WO2020200161A1 - Salt of indazolyl-containing tricyclic derivative and crystal form thereof - Google Patents

Salt of indazolyl-containing tricyclic derivative and crystal form thereof Download PDF

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WO2020200161A1
WO2020200161A1 PCT/CN2020/082042 CN2020082042W WO2020200161A1 WO 2020200161 A1 WO2020200161 A1 WO 2020200161A1 CN 2020082042 W CN2020082042 W CN 2020082042W WO 2020200161 A1 WO2020200161 A1 WO 2020200161A1
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acid
crystal form
pyridin
characteristic peaks
compound
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PCT/CN2020/082042
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French (fr)
Chinese (zh)
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詹小兰
呙临松
李宗斌
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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 CN202080002252.1A priority Critical patent/CN112020357B/en
Publication of WO2020200161A1 publication Critical patent/WO2020200161A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/4162—1,2-Diazoles condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5365—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/553—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having at least one nitrogen and one oxygen as ring hetero atoms, e.g. loxapine, staurosporine
    • 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
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains three hetero rings
    • C07D471/14—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the invention belongs to the field of drug synthesis, and specifically relates to a salt of an indazole group-containing triacyl derivative and a preparation method and application of its crystal form.
  • ERK signaling pathway Members of the ERK signaling pathway, such as RAS and BRAF, often mutate in tumors. About 1/3 of human tumors express continuously activated mutant RAS, and 8% of tumors express activated BRAF.
  • the mutations and probability statistics related to the ERK signaling pathway in malignant tumors are shown in Table 1. According to statistics, 90% of pancreatic cancers, 50% of colorectal cancers, and 30% of lung cancers have RAS mutations; 50% of melanomas, 50% of thyroid cancers, and 15% of colorectal cancers have BRAF mutations.
  • Vemurafenib is the first BRAF inhibitor approved by the FDA to be marketed. It is mainly used for the treatment of advanced melanoma, but the efficacy can only be maintained for 8-9 months, which is prone to drug resistance. Studies have confirmed that reactivation of the ERK signaling pathway mediates the resistance of melanoma to verofenil. And another BRAF inhibitor Dabrafenib (Dabrafenib) is also very easy to develop resistance. In addition, verofenib has failed to show significant clinical activity in colorectal cancer patients with BRAF mutations, and the overall response rate is only 5%. In addition to BRAF inhibitors, MEK inhibitors currently on the market have also appeared to varying degrees of resistance in clinical applications.
  • MEK inhibitors have a low response rate to RAS mutation tumors, and the response rate to BRAF mutation melanoma is only 22%.
  • BRAF inhibitors and EGFR inhibitors are used clinically to reverse drug resistance, and the patient develops multidrug resistance several months later.
  • PCT patent application number: PCT/CN2018/110795 discloses the structure of a series of pyrazolyl-containing triacyl derivative inhibitors.
  • PCT/CN2018/110795 discloses the structure of a series of pyrazolyl-containing triacyl derivative inhibitors.
  • the present invention has conducted a comprehensive study on the salts of the above substances, and is dedicated to obtaining the most suitable salt and crystal form for medicine.
  • the purpose of the present invention is to provide a crystal form of the compound represented by general formula (I), the structure of which is as shown in formula (I):
  • W is selected from N or CH
  • R 1 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R 3 , -(CH 2 ) n OR 3 , -(CH 2 ) n C(O)R 3 , -(CH 2 ) n NR 3 R 4 or -(CH 2 ) n C(O)NR 3 R 4 ;
  • R 2 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl or 3-8 membered heterocyclic group;
  • R 3 and R 4 are the same or different, and are each independently selected from hydrogen atom, deuterium atom, halogen, cyano, hydroxyl, amino, nitro, C 1-6 alkyl, C 1-6 haloalkyl, C 3- 8 cycloalkyl or 3-8 membered heterocyclic group, wherein the C 1-6 alkyl, C 1-6 haloalkyl, amino, C 3-8 cycloalkyl and 3-8 membered heterocyclic group are optional Further selected from deuterium atom, C 1-6 alkyl, C 1-6 haloalkyl, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl and 3-6 membered heterocyclic group substituted by one or more substituents;
  • M is an inorganic acid or an organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1- Hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-amino Benzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid,
  • x is selected from an integer of 0, 1, 2 or 3;
  • n is an integer of 0, 1, 2 or 3;
  • t is an integer of 0, 1, 2, 3, 4, 5, or 6.
  • the crystal form of the compound represented by general formula (I) has a structure as shown in general formula (II):
  • W, R 1 , R 2 , M, x and t are as defined in general formula (I).
  • the crystal form of the compound represented by general formula (I) has the following compound structure:
  • x and M are as defined in general formula (I).
  • it is the free base crystal form of the compound of formula (III).
  • it is an anhydrate, and further contains water, and the water is pipeline water or crystal water, preferably containing 0.5-8 water molecules; more preferably, the hydrate contains 0.5-4 water molecules; further It preferably contains 0.5-2.5 water molecules; more preferably, it contains 1 water molecule. For example, it contains 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 or 8 water.
  • the crystal form of the compound of formula (III) is a hydrate or an anhydrate.
  • the crystal form of the compound of formula (III) contains 0.5-8 water molecules; preferably, the hydrate contains 0.5-4 water molecules; further preferably, it contains 0.5-2.5 water molecules. Molecule; More preferably, it contains 1 water molecule. For example, it contains 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 or 8 water.
  • M is selected from maleic acid, p-toluenesulfonic acid, hydrochloric acid, nitric acid, sulfuric acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid or 1,5-naphthalenedisulfonic acid
  • x is selected from 0.5, 1, 1.5 or 2, preferably 1.
  • x and M are as defined in general formula (I).
  • Another object of the present invention is to provide a compound represented by general formula (Ia), the structure of which is as shown in formula (Ia):
  • W is selected from N or CH
  • R 1 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R 3 , -(CH 2 ) n OR 3 , -(CH 2 ) n C(O)R 3 , -(CH 2 ) n NR 3 R 4 or -(CH 2 ) n C(O)NR 3 R 4 ;
  • R 2 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl or 3-8 membered heterocyclic group;
  • R 3 and R 4 are the same or different, and are each independently selected from hydrogen atom, deuterium atom, halogen, cyano, hydroxyl, amino, nitro, C 1-6 alkyl, C 1-6 haloalkyl, C 3- 8 cycloalkyl or 3-8 membered heterocyclic group, wherein the C 1-6 alkyl, C 1-6 haloalkyl, amino, C 3-8 cycloalkyl and 3-8 membered heterocyclic group are optional Further selected from deuterium atom, C 1-6 alkyl, C 1-6 haloalkyl, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl and 3-6 membered heterocyclic group substituted by one or more substituents;
  • M is an inorganic acid or an organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1- Hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-amino Benzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid
  • y is an integer of 1, 2 or 3;
  • n is an integer of 0, 1, 2 or 3;
  • t is an integer of 0, 1, 2, 3, 4, 5, or 6.
  • the compound represented by general formula (Ia) has a structure as shown in formula (IIa):
  • R 1 , R 2 , W, M, t, and x are as defined in general formula (I).
  • the compound represented by the general formula (Ia) has a compound structure as shown in the formula (IIIa):
  • M and y are as defined in general formula (Ia).
  • the compound represented by the general formula (IIIa) is a water anhydride, further comprising water, and the water is pipeline water or crystal water.
  • the compound represented by the general formula (IIIa) contains 0.5-8 water molecules; preferably 0.5-4 water molecules; further preferably 0.5-2.5 water molecules; Preferably 1 water molecule; for example, it contains 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 or 8 water.
  • the compound represented by the general formula (Ia) has a compound structure as shown in formula (IVa):
  • M and y are as defined in general formula (Ia), preferably, M is selected from maleic acid, and y is 1.
  • the object of the present invention is also to provide a method for preparing the crystal form of the compound represented by general formula (I) or the compound represented by general formula (Ia), which specifically includes the following steps:
  • the benign solvent is selected from methanol, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidone; preferably N,N-dimethylformamide and N- Methylpyrrolidone;
  • the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol and ethanol; the above
  • the benign solvent and organic solution must be miscible when used;
  • the poor solvent is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably ethyl acetate, methyl tert-butyl ether, isopropyl ether and acetonitrile ;
  • the counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, py
  • the object of the present invention is also to provide a crystal form of the compound represented by general formula (I) or a method for preparing the compound represented by general formula (Ia), which specifically includes the following steps:
  • the poor solvent is selected from acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N , N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol or 2-butanone; preferably methanol, ethanol, tetrahydrofuran, ethyl acetate, acetonitrile or acetone; preferably methanol, Ethanol, tetrahydrofuran, ethyl acetate, acetonitrile and acetone;
  • the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol or ethanol; the above
  • the benign solvent and organic solution must be miscible when used;
  • the counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, py
  • the object of the present invention is also to provide a crystal form of the compound represented by general formula (I) or a method for preparing the compound represented by general formula (Ia), which specifically includes the following steps:
  • the benign solvent is selected from methanol, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidone; preferably N,N-dimethylformamide and N- Methylpyrrolidone;
  • the organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol and ethanol; the above
  • the benign solvent and organic solution must be miscible when used;
  • the counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroga
  • the crystalline form of the compound represented by the general formula (IV), the free base crystalline form I, the X-ray powder diffraction pattern is at 2 ⁇ of 6.4 and 26.7 (2 ⁇ 0.2 °) has characteristic peaks, preferably, it further includes characteristic peaks at 2 ⁇ of 7.2, 13.1, 16.8, 17.7, 18.9, 20.2, 21.2, and 28.2 (2 ⁇ 0.2°).
  • the crystal form of the compound represented by the general formula (IV) of the present invention which is a free base compound crystal form, uses Cu-K ⁇ radiation, and the X-ray characteristic diffraction peaks expressed in 2 ⁇ angles and interplanar spacing d are shown in the table 1 shown.
  • the crystalline form of the compound represented by the general formula (IV) of the present invention is the crystalline form I of the free base compound, and its X-ray powder diffraction pattern is basically shown in FIG. 1.
  • the crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form I of the free base compound, and its TGA pattern is basically shown in FIG. 2.
  • the crystalline form of the compound represented by the general formula (IV) of the present invention is the crystalline form I of the free base compound, and its DSC spectrum is basically shown in FIG. 3.
  • the crystalline form of the compound represented by the general formula (IV), its free base crystalline form II, the X-ray powder diffraction pattern is at 2 ⁇ of 8.5, 13.4 and 17.0 (2 ⁇ ⁇ 0.2°) has characteristic peaks, and also includes characteristic peaks at 2 ⁇ of 10.2, 10.9, 12.8, 13.1, 16.1, 17.8, 18.8, 19.5, 23.0, 23.9, 24.4, 25.5, 26.1 and 27.6 (2 ⁇ 0.2°) .
  • the crystal form of the compound represented by the general formula (IV) of the present invention which is a free base compound crystal form, uses Cu-K ⁇ radiation, and the X-ray characteristic diffraction peaks expressed in 2 ⁇ angles and interplanar spacing d are shown in the table 2 shown.
  • the crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form II of the free base compound, and its X-ray powder diffraction pattern is basically as shown in FIG. 4.
  • the crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form II of the free base compound, and its TGA pattern is basically shown in FIG. 5.
  • the crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form II of the free base compound, and its DSC spectrum is basically as shown in FIG. 6.
  • the powder diffraction pattern has characteristic peaks at 2 ⁇ of 5.5 and 16.3 (2 ⁇ 0.2°); it further includes characteristic peaks at 2 ⁇ of 10.8, 15.3, 17.7, 26.1, 26.4 and 27.0 (2 ⁇ 0.2°); further There are characteristic peaks at 2 ⁇ of 13.0, 14.9, 20.0 and 20.8 (2 ⁇ 0.2°);
  • the crystal form of the compound represented by the general formula (IV) of the present invention wherein M is maleic acid, and x is 1, and the maleate salt crystal form I, using Cu-K ⁇ radiation, is set at a 2 ⁇ angle and a crystal plane
  • the X-ray characteristic diffraction peaks represented by the spacing d are shown in Table 3.
  • the X-ray powder diffraction pattern is basically shown in Figure 7.
  • the DSC spectrum is basically shown in Figure 9.
  • the X-ray powder diffraction pattern has characteristic peaks at 2 ⁇ of 5.5, 10.8, 16.3 and 17.6 (2 ⁇ 0.2°); it further contains characteristic peaks at 2 ⁇ of 14.0, 26.4, 26.6 and 27.2; There are characteristic peaks at 8.8, 12.9, 18.4, 18.9, 20.6, 21.5, 22.4 and 22.8 (2 ⁇ 0.2°).
  • the X-ray characteristic diffraction peaks represented by the spacing d are shown in Table 4.
  • the X-ray powder diffraction pattern is basically shown in Figure 10.
  • the powder diffraction pattern has characteristic peaks at 2 ⁇ of 4.9, 15.1, 17.2, 17.5, 26.5 and 26.9 (2 ⁇ 0.2°); it further contains characteristic peaks at 2 ⁇ of 9.8, 14.7, 18.4 and 19.8 (2 ⁇ 0.2°) Peak; further includes characteristic peaks at 2 ⁇ of 12.3, 13.0, 14.0, and 27.5 (2 ⁇ 0.2°).
  • the powder diffraction pattern is basically shown in Figure 12.
  • the crystal form of the compound represented by formula (IV), wherein M is maleic acid, and x is 1, its maleate salt crystal form III, and its DSC spectrum is basically As shown in Figure 13.
  • the spectrum has characteristic peaks of 4.8 and 16.7 in 2 ⁇ ; it further includes characteristic peaks at 2 ⁇ of 9.6, 11.9 and 14.9 (2 ⁇ 0.2°); it is further included in 2 ⁇ of 7.2, 12.9, 14.3, 19.1, 19.3, There are characteristic peaks at 21.5 and 23.9 (2 ⁇ 0.2°).
  • the powder diffraction pattern is basically shown in Figure 14.
  • the crystal form of the compound represented by formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form IV, and its DSC spectrum is basically as shown in Figure 15.
  • the crystal form of the compound represented by formula (IV), wherein M is hydrochloric acid, the hydrochloride crystal form, and the X-ray powder diffraction pattern is at 2 ⁇ of 5.8 and 17.2 (2 ⁇ 0.2°) has characteristic peaks; further includes characteristic peaks at 2 ⁇ of 12.0, 13.6, 16.4, 21.7, 23.0, 26.1, 26.3, and 27.1 (2 ⁇ 0.2°); it is further included in 2 ⁇ of 8.9, There are characteristic peaks at 28.8 and 30.1 (2 ⁇ 0.2°).
  • the crystal form of the compound represented by formula (IV), wherein M is hydrochloric acid, its hydrochloride crystal form, and its X-ray powder diffraction pattern is basically as shown in Figure 16. .
  • M is p-toluenesulfonic acid
  • its p-toluenesulfonic acid salt crystal form I X-ray powder diffraction
  • the spectrum has characteristic peaks at 2 ⁇ of 6.1, 11.3, 17.0, 17.5 and 18.3 (2 ⁇ 0.2°); it further contains characteristic peaks at 2 ⁇ of 12.7, 19.1, 19.9, 20.6 and 22.2 (2 ⁇ 0.2°).
  • the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form I, and its X-ray powder is basically shown in Figure 17.
  • M is p-toluenesulfonic acid
  • its p-toluenesulfonic acid salt crystal form II X-ray powder diffraction
  • the spectrum has characteristic peaks at 2 ⁇ of 5.4, 8.6, 11.4, 16.8, 18.2, 19.7, 20.4, and 21.9 (2 ⁇ 0.2°); it is further included at 2 ⁇ of 13.5, 13.7, 15.7, 17.2, 23.7, 25.5 and 27.6 (2 ⁇ 0.2°) has characteristic peaks; and further includes characteristic peaks at 2 ⁇ of 16.0, 21.4, 25.9, and 28.7 (2 ⁇ 0.2°).
  • the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form II, and its X-ray powder is basically shown in Figure 18.
  • the spectrum has characteristic peaks at 2 ⁇ of 4.9, 8.6, 13.2, 18.9, 20.6, and 25.2 (2 ⁇ 0.2°); it is further included at 2 ⁇ of 9.6, 10.9, 12.6, 15.0, 15.6, 17.0, 22.6, 25.8, and 27.5. (2 ⁇ 0.2°) has characteristic peaks; and further includes characteristic peaks at 2 ⁇ of 10.5, 13.9, 16.5, 17.7, 21.7, 26.1, 26.6, 26.9, 27.8, 29.8, and 32.2 (2 ⁇ 0.2°).
  • the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form III, and its X-ray powder is basically shown in Figure 19.
  • M is p-toluenesulfonic acid
  • its p-toluenesulfonic acid salt crystal form IV X-ray powder diffraction
  • the spectrum has characteristic peaks at 2 ⁇ of 5.5, 11.9, 16.3, 19.4, and 25.5 (2 ⁇ 0.2°); it further includes 2 ⁇ at 13.8, 18.1, 18.6, 20.1, 21.3, 22.9, and 26.5 (2 ⁇ 0.2°) It has characteristic peaks; it further includes characteristic peaks at 8.7, 9.1, 10.2, 10.8, 12.9 and 20.4 (2 ⁇ 0.2°).
  • the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form IV, and its X-ray powder is basically shown in Figure 20.
  • the crystal form of the compound represented by formula (IV), wherein M is nitric acid, its nitrate crystal form, and the X-ray powder diffraction pattern at 2 ⁇ is 5.0, 16.3, 16.7 And 28.0 (2 ⁇ 0.2°) with characteristic peaks; further include characteristic peaks at 2 ⁇ of 8.2, 8.5, 11.8, 13.2, and 29.1 (2 ⁇ 0.2°); further including 2 ⁇ at 19.7, 20.4, 21.0 And 24.5 (2 ⁇ 0.2°) have characteristic peaks.
  • the crystal form of the compound represented by formula (IV), wherein M is nitric acid, its nitrate crystal form, and its X-ray powder diffraction pattern is basically as shown in FIG. 21.
  • the object of the present invention is also to provide a pharmaceutical composition, which contains a therapeutically effective amount of the crystal form of the compound of formula (I) or the compound of formula (Ia), and one or more A pharmaceutically acceptable carrier.
  • the purpose of the present invention is also to provide the crystal form of the general formula compound represented by formula (I) or the general formula compound represented by formula (Ia) and the pharmaceutical composition in the preparation for the treatment and/or prevention of ERK-mediated The use of drugs for leading cancer or tumor-related diseases.
  • the object of the present invention is also to provide the crystal form of the compound represented by formula (I) or the compound represented by formula (Ia) and its pharmaceutical composition for preparing and treating cancer, inflammation, chronic liver disease, diabetes, heart disease Application in medicine for vascular disease or AIDS.
  • the object of the present invention is also to provide a method for the treatment, prevention and/or treatment and prevention of ERK-mediated pathological characteristics of diseases, which comprises administering to a patient a therapeutically effective dose of a crystal of a compound of formula (I) or Compounds of general formula represented by formula (Ia) and pharmaceutical compositions thereof.
  • Diseases with ERK-mediated pathological characteristics include cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease or AIDS.
  • the disease is selected from cancer, bone disease, inflammatory disease, immune disease, neurological disease, metabolic disease, respiratory disease, or medical application in heart disease; wherein the cancer is selected from Breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, acute myeloid leukemia, or colorectal cancer.
  • 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
  • the alkyl group is 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
  • Alkyl groups may be substituted or unsubstituted. When substituted, the substituents may be substituted at any available attachment point.
  • the substituents are preferably one or more of the following groups, which are independently selected from alkanes Group, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkane Oxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate group, the present invention preferably methyl, ethyl, isopropyl, tert-butyl, haloalkyl , 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” It refers to -(CH 2 ) 3 -, “butylene” refers to -(CH 2 ) 4 -, etc.
  • the above-mentioned substituents can be connected to different carbon atoms to form a carbon chain, or they can be connected to a carbon atom to form a cycloalkyl group.
  • alkenyl refers to an alkyl group as defined above composed of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3 -Butenyl etc. Alkenyl groups may be substituted or unsubstituted.
  • the substituents are preferably one or more of the following groups, which are independently selected from 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 contains 3 to 20 carbon atoms, preferably 3 to 8 carbon atoms, more preferably 3 to 6 Carbon atoms.
  • Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene
  • Polycyclic cycloalkyl groups include spiro, condensed and bridged cycloalkyl groups, preferably cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl and cycloheptyl.
  • heterocyclyl refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent which contains 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (where m is an integer of 0 to 2) heteroatoms, but does not include the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably contains 3 to 8 ring atoms; most preferably contains 3 to 8 ring atoms.
  • Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidine Group, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably tetrahydrofuryl, pyrazolidinyl, morpholinyl, piperazinyl and pyranyl.
  • Polycyclic heterocyclic groups include spiro, condensed and bridged heterocyclic groups; the spiro, condensed and bridged heterocyclic groups are optionally connected to other groups through a single bond, or through a ring Any two or more of the above atoms are further connected to other cycloalkyl groups, heterocyclic groups, aryl groups and heteroaryl groups; heterocyclic groups may be optionally substituted or unsubstituted, when substituted ,
  • the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano , Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy,
  • heterocyclyl refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent which contains 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (where m is an integer of 0 to 2) heteroatoms, but does not include the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably contains 3 to 8 ring atoms; most preferably contains 3 to 8 ring atoms.
  • Non-limiting examples of monocyclic heterocyclic groups include oxetanyl, pyrrolidinyl, pyrrolidone, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrrolidinyl Azolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably oxetanyl, pyrrolidonyl, tetrahydrofuranyl, Pyrazolidinyl, morpholinyl, piperazinyl and pyranyl.
  • Polycyclic heterocyclic groups include spiro, condensed and bridged heterocyclic groups; the spiro, condensed and bridged heterocyclic groups are optionally connected to other groups through a single bond, or through a ring Any two or more of the above atoms are further connected to other cycloalkyl groups, heterocyclic groups, aryl groups and heteroaryl groups.
  • alkoxy refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), where alkyl is defined as described above. It is preferably an alkoxy group having 1 to 8 carbon atoms, more preferably an alkoxy group having 1 to 6 carbon atoms, and most preferably an alkoxy group having 1 to 3 carbon atoms.
  • alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy.
  • the alkoxy group may be optionally substituted or unsubstituted.
  • the substituent is preferably one or more of the following groups, which are independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , Heterocycloalkylthio, carboxyl or carboxylate.
  • Haloalkyl refers to an alkyl group substituted with one or more halogens, where alkyl is as defined above.
  • Haloalkoxy refers to an alkoxy group substituted with one or more halogens, where alkoxy is as defined above.
  • Hydroalkyl refers to an alkyl group substituted with a hydroxy group, where the alkyl group is as defined above.
  • Alkenyl refers to an alkenyl group, also known as an alkene group, preferably an alkenyl group containing 2 to 8 carbon atoms, more preferably an alkenyl group having 2 to 6 carbon atoms, and most preferably an alkenyl group having 2 to 3 carbon atoms ;
  • the alkenyl group can be further substituted by other related groups, such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano , Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.
  • Alkynyl refers to (CH ⁇ C-), preferably an alkynyl group containing 2 to 8 carbon atoms, more preferably an alkynyl group having 2 to 6 carbon atoms, and most preferably an alkynyl group having 2 to 3 carbon atoms.
  • alkynyl group described therein may be further substituted by other related groups, such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy, or carboxylate.
  • other related groups such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy, or carboxylate.
  • Halogen refers to fluorine, chlorine, bromine or iodine.
  • Amino refers to -NH 2 .
  • Cyano refers to -CN.
  • Niro refers to -NO 2 .
  • Carboxy refers to -C(O)OH.
  • 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
  • other terms all express the same Meaning, which means that X can be any one or more of A, B, and C.
  • Substituted refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of each other, substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without too much effort. For example, an amino group or a hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (eg, olefinic) bond.
  • Stepoisomerism includes geometric isomerism (cis-trans isomerism), optical isomerism, and conformational isomerism.
  • the hydrogen atoms described in the present invention can be replaced by its isotope deuterium, and any hydrogen atom in the example compounds of the present invention can also be replaced by a deuterium atom.
  • “Pharmaceutical composition” means a mixture containing one or more of the compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, and other components such as physiological/pharmaceutically acceptable carriers And excipients.
  • the purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredients and thus the biological activity.
  • “Pharmaceutically acceptable salt” refers to the salt of the compound of the present invention. Such salt is safe and effective when used in mammals, and has due biological activity.
  • TGA thermogravimetric analysis
  • DSC differential scanning calorimetry
  • DVD refers to dynamic moisture adsorption (DVS) experiments.
  • XRPD refers to X-ray powder diffraction (XRPD) experiments.
  • HPLC refers to high performance liquid chromatography (HPLC) experiments.
  • PK refers to pharmacokinetic (PK) experiments.
  • Figure 1 is an XRPD diagram of the free base crystal form I.
  • Figure 2 is a TGA diagram of the free base crystal form I.
  • Figure 3 is a DSC diagram of the free base crystal form I.
  • Figure 4 is an XRPD diagram of the free base crystal form II.
  • Figure 5 is a TGA diagram of the free base crystal form II.
  • Figure 6 is a DSC chart of the free base crystal form II.
  • Figure 7 is an XRPD diagram of maleate salt form I.
  • Figure 8 is a TGA diagram of maleate salt form I.
  • Figure 9 is a DSC diagram of maleate salt form I.
  • Figure 10 is an XRPD diagram of maleate salt form II.
  • Figure 11 is a DSC chart of maleate salt form II.
  • Figure 12 is an XRPD diagram of maleate salt form III.
  • Figure 13 is a DSC chart of maleate salt form III.
  • Figure 14 is an XRPD diagram of maleate salt form IV.
  • Figure 15 is a DSC chart of maleate salt form IV.
  • Figure 16 is an XRPD diagram of the hydrochloride salt.
  • Figure 17 is an XRPD diagram of p-toluenesulfonate form I.
  • Figure 18 is an XRPD diagram of p-toluenesulfonate Form II.
  • Figure 19 is an XRPD diagram of p-toluenesulfonate Form III.
  • Figure 20 is an XRPD diagram of p-toluenesulfonate salt form IV.
  • Figure 21 is an XRPD diagram of nitrate.
  • Figure 22 is a schematic diagram of a rat PK experiment.
  • Mobile phase A: water (0.05% trifluoroacetic acid); B: acetonitrile (0.05% trifluoroacetic acid)
  • the first step preparation of methyl 3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)acryloyl acid
  • reaction solution is concentrated and purified by column chromatography [eluent: petroleum ether ⁇ petroleum ether/ethyl acetate (60:40)] to obtain a pale blue solid product 3-(6-nitro-3-(pyridine-4) -Yl)-1-tritylmethyl-1H-indazol-5-yl)acrylic acid methyl ester (200 mg, yield 51%).
  • reaction solution was concentrated and purified by column chromatography [eluent: petroleum ether ⁇ petroleum ether/ethyl acetate (40:60)] to obtain the product (R)-N-(1-phenylethyl)-3- (Pyridin-4-yl)-1-tritylmethyl-1,7-dihydropyrrolo[3,2-f]indazole-6-carboxamide (50 mg, yield 40%).
  • the third step Preparation of 3-(pyridin-4-yl)-1-trityl-1H-indazole-5,6-diamine
  • Step 4 Preparation of 3-(pyridin-4-yl)-6-(trichloromethyl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole
  • Step 5 Preparation of 3-(pyridin-4-yl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester
  • Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate Place in a microwave reaction tube, add (R)-1-(p-benzyl)ethane-1-amine (1.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution.
  • Step 2 Preparation of N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
  • N-(1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6- The crude formamide (27mg) was dissolved in dichloromethane (2mL), added with trifluoroacetic acid (2mL), stirred at room temperature for 2h, concentrated to dryness, purified by column chromatography to obtain a yellow solid N-(1-phenylethyl)-3 -(Pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (5 mg, yield 7% in two steps).
  • Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate Place in a microwave reaction tube, add (R)-1-(4-(trifluoromethyl)phenyl)ethane-1-amine (1.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution.
  • the obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried over anhydrous sodium sulfate.
  • the obtained organic solvent was spin-dried and purified by preparative chromatography to obtain the yellow solid product (R)-3-(pyridin-4-yl)-N-(1-(3-(trifluoromethyl)phenyl)ethyl)- 1,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide (10 mg, yield 19%).
  • Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate Place in a microwave reaction tube, add (R)-1-(3-fluorophenyl)ethane-1-amine (1.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution.
  • the third step Preparation of 6-fluoro-3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine
  • Step 7 Preparation of methyl 3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-b]pyrazolo[4,3-e]pyridine-6-carboxylate
  • the first step preparation of (R)-(1-(3-bromophenyl)ethyl) t-butyl carbamate
  • Step 2 Preparation of tert-butyl (R)-(1-(3-cyclopropylphenyl)ethyl)carbamate
  • reaction solution is concentrated and purified by column chromatography [eluent: petroleum ether ⁇ petroleum ether/ethyl acetate (95/5)] to obtain a colorless oily product (R)-(1-(3-cyclopropylbenzene) (Yl)ethyl) tert-butyl carbamate (430 mg, yield 69%).
  • Step 2 (R)-N-(1-(3-(Pro-1-en-2-yl)phenyl)ethyl)-3-(pyridin-4-yl)-1-trityl Of phenyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
  • reaction solution was concentrated and purified by column chromatography [eluent: petroleum ether ⁇ petroleum ether/ethyl acetate (30:70)] to obtain a brown oily product (R)-N-(1-(3-(prop-1-ene) -2-yl)phenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide (100 mg, 53%).
  • test case is to measure the inhibitory ability of the compound on ERK-1 kinase activity.
  • In vitro ERK-1 kinase analysis was performed using the LANCE Ultra (Perkin Elmer) method.
  • ERK1 enzyme Invitrogen, #PV3311, final concentration: 0.4nM
  • kinase buffer 50mM Hepes pH 7.4, 10mM MgC12, 1mM EGTA, 0.01% Triton X-100, 2mM DTT
  • substrate Ulight-MBP peptide Perkin
  • test case The purpose of this test case is to measure the compound's ability to inhibit ERK-2 kinase activity.
  • In vitro ERK-2 kinase analysis was performed using LANCE Ultra (Perkin Elmer) method. By adding 2.5 ⁇ L of test compound/DMSO (final 4%, V/V, diluted to 10 concentrations (400nM to 0.02nM) using a 1:3 dilution scheme) in a 384-well plate (Perkin Elmer OPTIPLATE TM ), adding ERK-2 enzyme (Invitrogen, #PV3313, final concentration: 0.08nM) prepared in kinase buffer (50mM Hepes pH 7.4, 10mM MgCl 2 , 1mM EGTA, 0.01% Triton X-100, 2mM DTT) and substrate Ulight-MBP Peptide (Perkin Elmer, #TRF0109-M, final concentration: 0.5 ⁇ M) mixed solution 5 ⁇ L, then add 2.5 ⁇ L of ATP (
  • the compound shown in the present invention shows a biological activity of about 0.01 nM to 100 nM (IC 50 ) in the ERK inhibition test.
  • compounds of the invention and for the IC ERK-1 / ERK-2 is 50 or less than about 10OnM, preferably less than about 10 nM compound, more preferably less than about of 5 nM, more preferably less than about 1 nM, the present invention is presented The most preferred compound is less than 0.1 nM.
  • compounds of the invention for the ERK IC 50 less than about 10OnM, preferably less than about 10 nM, more preferably less than about of 5 nM, more preferably less than about 1 nM, most preferably less than compounds listed 0.1nM.
  • the present invention is preferably a compound listed compounds show dual binding specificity, and can be less than about 10OnM, less than about 10 nM, less than about of 5 nM, less than about 1 nM, less than the IC 50 values of 0.1nM of ERK Kinases (eg, ERK-1 kinase, ERK-2 kinase, etc.) and protein kinases (eg, Ras, Raf, Her-2, MEK1, etc.).
  • ERK Kinases eg, ERK-1 kinase, ERK-2 kinase, etc.
  • protein kinases eg, Ras, Raf, Her-2, MEK1, etc.
  • test data of the embodiment is shown in Table 13:
  • the purpose of this test example is to determine the inhibitory effect of the compound of the present invention on tumor cell proliferation activity.
  • the compound's anti-tumor cell proliferation inhibitory activity was measured by the CellTiter-Glo method, and the half inhibitory concentration IC 50 of the compound's inhibitory cell proliferation activity was obtained.
  • the test compound solutions of different concentrations in gradient dilutions are added to the cells of the culture plate, and the culture plate is incubated in an incubator for 3-7 days (37° C., 5% CO 2 ).
  • the compounds of the present invention were tested for the proliferation activity of pancreatic cancer tumor cells Mia Paca 2, and the measured IC 50 values are shown in Table 14.
  • Table 14 Relative IC 50 value of compound inhibiting proliferation activity of pancreatic cancer tumor cell Mia Paca 2
  • the compound of the present invention has obvious inhibitory effect on tumor cell proliferation activity.
  • Balb/c Mouse male, purchased from Shanghai Jiesjie Laboratory Animal Co., Ltd., animal production license number (SCXK (Shanghai) 2013-0006N0.311620400001794) was used as the test animal to study the compound
  • Example 1 The pharmacokinetic behavior of Example 1, Example 2, and Example 6 in Mouse (plasma).
  • Example 1 Weigh 11.2mg of Example 1, 10.8mg of Example 2, and 9.8mg of Example 6, respectively into 4mL glass bottles, add 1.940mL, 2.160mL and 1.525mL of this solution, sonicate for 10 minutes to obtain a colorless clear solution with a concentration of 5mg/mL. After fasting overnight, PO respectively, the dose was 50 mg/kg, and the administration volume was 10 mL/kg.
  • mice BALB/c nude mice were used as test animals, and the human pancreatic cancer cell MiaPaca 2 xenograft (CDX) model was used for in vivo pharmacodynamic experiments to evaluate the anti-tumor effects of the test compounds.
  • CDX human pancreatic cancer cell MiaPaca 2 xenograft
  • Vernier caliper (CD-6"AX, Mitutoyo, Japan)
  • FBS Fetal Bovine Serum
  • Penicillin double antibody (SV30010, GE)
  • PBS Phosphate buffered saline
  • MiaPaca 2 cells Take out MiaPaca 2 cells from the cell bank, add DMEM medium (containing 10% FBS, 1% Glu, 1% P/S) after resuscitation and place them in a CO 2 incubator (the temperature of the incubator is 37°C, and the CO 2 concentration Is 5%). After the cells are 80-90% of the bottom of the culture flask, the cells are passaged. After passage, the cells are continued to be cultured in a CO 2 incubator. Repeat the process until the number of cells meets the requirement of inoculation in vivo, start to collect the cells in the logarithmic growth phase, count them with an automatic cell counter, and resuspend the cells with PBS and Matrigel (volume ratio 1:1) according to the counting results. Prepare cell suspension (density 8 suspended cells 7 /ml) and place in an ice box for later use.
  • DMEM medium containing 10% FBS, 1% Glu, 1% P/S
  • mice Female, 6-8 weeks old, weighing about 18-22 grams.
  • the mice were kept in an environment free of special pathogens and in a single ventilated cage with 5 mice per cage. All cages, litter and water are disinfected before use, and all animals can freely access standard certified commercial laboratory food.
  • the nude mice were labeled with disposable universal ear tags for rats and mice, and the skin of the inoculation site was disinfected with 75% medical alcohol before inoculation.
  • Each mouse was inoculated with 0.1ml (containing 8*10 6 cells) MiaPaca 2 subcutaneously on the right back of each mouse. Tumor cells. When the average tumor volume reaches 100-200mm 3 , group administration is started.
  • the test compound was administered by oral gavage every day, and the dosage, frequency of administration and the efficacy of each group at the end of the experiment are shown in Table 16 and Table 17.
  • the antitumor efficacy is determined by dividing the average tumor increase volume of animals treated with the compound by the average tumor increase volume of untreated animals.
  • Example 2 can significantly inhibit the growth of transplanted tumors in nude mice of MiaPaca 2 under the conditions of 100mg/kg QD and 50mg/kg BID, and the drug effect is good;
  • Example 6 100mg/kg QD can also significantly inhibit tumor growth.
  • Example 1 can significantly inhibit the growth of transplanted tumors in MiaPaca 2 nude mice under the conditions of 50 mg/kg QD administration, and the drug effect is good.
  • CHO-hERG cells are cultured in a 175cm2 culture flask. After the cell density grows to 60-80%, remove the culture solution, wash it with 7mL PBS, and then add 3mL Detachin for digestion.
  • the single-cell high-impedance sealing and the whole-cell mode formation process are all automatically completed by the Qpatch instrument.
  • the cell is clamped at -80 mV, before giving a 5 second +40 mV depolarization stimulus .
  • This voltage stimulus was applied every 15 seconds, recorded for 2 minutes, and then administered extracellular fluid for 5 minutes, and then the administration process was started.
  • the compound concentration started from the lowest test concentration, and each test concentration was given for 2.5 minutes. At least 3 cells are tested at each concentration.
  • the highest test concentration is 40uM, which are respectively 40, 13.33, 4.44, 1.48, 0.49, 0.16uM and 6 concentrations.
  • the experimental data is analyzed by XLFit software.
  • the experimental reagents used were purchased from Sigma, with a purity of >98%
  • Example 2 has no inhibitory effect on the cardiac hERG potassium ion channel, and can avoid cardiac toxic side effects at high doses.
  • the present invention provides a series of highly active and selective ERK1/2 kinase inhibitors with novel structures. It has shown good pharmacokinetic properties in both rat and mouse models, and has also shown good pharmacodynamics in the Miapaca tumor-bearing mouse model. It has great potential to be developed for tumor diseases. drug.
  • Example 2 Weigh 500 mg of the free base of Example 2 and add 5 mL of N,N-dimethylformamide ultrasonically undissolved. The system is heated at 50°C until it is completely dissolved. Separately weigh 183 mg of maleic acid and 1 mL of methanol to completely dissolve at room temperature. clear. The methanol solution of maleic acid was added to the alkali solution at 50°C, and the precipitate precipitated out quickly. After stirring for 0.5h, 10mL methyl tert-butyl ether was added to the system. After stirring for 1.5h, the heating was turned off and the temperature was cooled to room temperature.
  • the filter cake is rinsed with 20 mL of methyl tert-butyl ether, and the solid is placed in a vacuum drying oven at 40° C. and dried to a constant weight to obtain maleate crystal form I. After detection and analysis, it has an XRPD chart as shown in FIG. 7, a TGA chart as shown in FIG. 8 and a DSC chart as shown in FIG. 9.
  • Maleate crystal form I absorbs moisture and increases weight by 1.36% under the condition of RH80%, which is slightly hygroscopic; after a cycle of moisture absorption and desorption under the condition of 0-95% relative humidity, maleate crystal form I
  • the XRPD spectrum has not changed, that is, there is no crystal transformation.
  • thermodynamic solubility of the compound at room temperature was determined by HPLC and external standard method.
  • maleate crystal form I weigh 50.23mg maleate crystal form I, add 0.75mL N-methylpyrrolidone to completely dissolve it at 50°C, add 1mL isopropyl ether to the system, and quickly precipitate a yellow solid. After reacting at 50°C for 2h Continue to add 2 mL of isopropyl ether and turn off the heating to bring to room temperature. Finally, the solid is centrifuged, and the supernatant is removed, and the solid is placed in a vacuum drying oven at 40° C. and dried to a constant weight to obtain maleate crystal form IV.
  • Maleate salt crystal form I is relatively stable in acetone, acetonitrile, ethanol, ethyl acetate, isopropanol and water.
  • Comparative compound example 2 has a free base crystal form II T max , and maleate salt crystal form I has a reduced T max .

Abstract

The present invention relates to a salt of an indazolyl-containing tricyclic derivative and a crystal form thereof. In particular, the present invention relates to a crystal form of a compound of general formula (I), a preparation method and a pharmaceutical composition containing a therapeutically effective amount of the crystal form, and the use of same in the preparation of a drug for treating related ERK-mediated diseases.

Description

含吲唑基的三并环类衍生物的盐及其晶型Salts and crystal forms of triacyl derivatives containing indazole groups 技术领域Technical field
本发明属于药物合成领域，具体涉及一种含吲唑基的三并环类衍生物的盐及其晶型的制备方法和应用。The invention belongs to the field of drug synthesis, and specifically relates to a salt of an indazole group-containing triacyl derivative and a preparation method and application of its crystal form.
背景技术Background technique
ERK信号通路的成员RAS、BRAF等在肿瘤中常常发生突变，大约有1/3人类肿瘤表达持续激活的突变型RAS，8％的肿瘤表达激活型的BRAF。恶性肿瘤中ERK信号通路相关的突变及概率统计见表1。据统计，90％的胰腺癌、50％的结直肠癌和30％肺癌的都存在RAS突变；50％的黑色素瘤、50％的甲状腺癌和15％的结直肠癌都有BRAF突变。Members of the ERK signaling pathway, such as RAS and BRAF, often mutate in tumors. About 1/3 of human tumors express continuously activated mutant RAS, and 8% of tumors express activated BRAF. The mutations and probability statistics related to the ERK signaling pathway in malignant tumors are shown in Table 1. According to statistics, 90% of pancreatic cancers, 50% of colorectal cancers, and 30% of lung cancers have RAS mutations; 50% of melanomas, 50% of thyroid cancers, and 15% of colorectal cancers have BRAF mutations.
表1.不同癌症中ERK信号通路相关突变类型及概率Table 1. Types and probabilities of mutations related to the ERK signaling pathway in different cancers
Figure PCTCN2020082042-appb-000001
Figure PCTCN2020082042-appb-000001
维罗非尼(Vemurafenib)是被FDA批准上市的第一个BRAF抑制剂，主要用于晚期黑色素瘤的治疗，但疗效仅可维持8-9个月，易发生耐药。研究证实重新激活ERK信号通路介导黑色素瘤对维罗非尼的耐药。而另一个BRAF抑制剂达拉非尼(Dabrafenib)也非常容易产生耐药。除此之外，维罗非尼在携带BRAF突变的结直肠癌病人中亦未能表现出显著的临床活性，总体响应率仅为5％。除了BRAF抑制剂以外，目前已经上市的MEK抑制剂在临床应用中也出现了不同程度的耐药。MEK抑制剂对RAS突变的肿瘤响应率低，对BRAF突变的黑色瘤响应率也仅为22％。临床采用BRAF抑制剂与EGFR抑制剂联用来逆转耐药，患者在数月后出现了多药耐药。Vemurafenib is the first BRAF inhibitor approved by the FDA to be marketed. It is mainly used for the treatment of advanced melanoma, but the efficacy can only be maintained for 8-9 months, which is prone to drug resistance. Studies have confirmed that reactivation of the ERK signaling pathway mediates the resistance of melanoma to verofenil. And another BRAF inhibitor Dabrafenib (Dabrafenib) is also very easy to develop resistance. In addition, verofenib has failed to show significant clinical activity in colorectal cancer patients with BRAF mutations, and the overall response rate is only 5%. In addition to BRAF inhibitors, MEK inhibitors currently on the market have also appeared to varying degrees of resistance in clinical applications. MEK inhibitors have a low response rate to RAS mutation tumors, and the response rate to BRAF mutation melanoma is only 22%. BRAF inhibitors and EGFR inhibitors are used clinically to reverse drug resistance, and the patient develops multidrug resistance several months later.
目前已经有大量的临床前研究结果证明不同种类的ERK上游靶点抑制剂耐药可以通过抑制ERK活性来逆转BRAF抑制剂和MEK抑制剂的耐药，已有很多制药公司在进行研究，如：Genentech、Merck、Lilly等，但目前尚未有中国本 土研究的BRAF及MEK抑制剂上市。现已公开的选择性抑制ERK1/2的专利申请包括WO2012088314、WO2014134776、WO2014179154、WO2014137728和WO2015051314等。但现有的临床候选药物如KO-947，存在溶解性差的问题，临床采用静脉给药，在常规溶剂中只能以混悬状态存在，不利于成药，更难以口服给药。At present, a large number of pre-clinical studies have proved that the resistance of different types of ERK upstream target inhibitors can reverse the resistance of BRAF inhibitors and MEK inhibitors by inhibiting ERK activity. Many pharmaceutical companies have been conducting research, such as: Genentech, Merck, Lilly, etc., but currently there are no domestically researched BRAF and MEK inhibitors on the market. Patent applications that have been published for selective inhibition of ERK1/2 include WO2012088314, WO2014134776, WO2014179154, WO2014137728 and WO2015051314. However, the existing clinical candidate drugs such as KO-947 have the problem of poor solubility. They are administered intravenously in clinical practice and can only exist in a suspension state in conventional solvents, which is not conducive to the preparation of medicines and is more difficult to administer orally.
PCT专利(申请号：PCT/CN2018/110795)中公开了一系列含吡唑基的三并环类衍生物抑制剂的结构，在后续的研发中，为了产物易于处理、过滤和干燥，寻求适合的便于储存、产品长期稳定等特点，本发明对上述物质的盐进行了全面的研究，致力于得到最适合成药的盐和晶型。The PCT patent (application number: PCT/CN2018/110795) discloses the structure of a series of pyrazolyl-containing triacyl derivative inhibitors. In the subsequent research and development, in order to facilitate the processing, filtration and drying of the product, a suitable Because of its convenient storage and long-term product stability, the present invention has conducted a comprehensive study on the salts of the above substances, and is dedicated to obtaining the most suitable salt and crystal form for medicine.
发明内容Summary of the invention
PCT专利(申请号：PCT/CN2018/110795)的内容均可以引证至本发明中，作为本申请的一部分。The content of the PCT patent (application number: PCT/CN2018/110795) can be cited in the present invention as a part of this application.
本发明的目的在于提供一种通式(I)所示化合物的晶型，其结构如式(I)所示：The purpose of the present invention is to provide a crystal form of the compound represented by general formula (I), the structure of which is as shown in formula (I):
Figure PCTCN2020082042-appb-000002
Figure PCTCN2020082042-appb-000002
其中：among them:
W选自N或CH；W is selected from N or CH;
R 1选自氢原子、卤素、氨基、硝基、羟基、氰基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6卤代烷氧基、C 1-6羟烷基、C 3-8环烷基、-(CH 2) nR 3、-(CH 2) nOR 3、-(CH 2) nC(O)R 3、-(CH 2) nNR 3R 4或-(CH 2) nC(O)NR 3R 4； R 1 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R 3 , -(CH 2 ) n OR 3 , -(CH 2 ) n C(O)R 3 , -(CH 2 ) n NR 3 R 4 or -(CH 2 ) n C(O)NR 3 R 4 ;
R 2选自氢原子、卤素、氨基、硝基、羟基、氰基、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卤代烷氧基、C 3-8环烷基或3-8元杂环基； R 2 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl or 3-8 membered heterocyclic group;
R 3和R 4相同或不同，且各自独立地选自氢原子、氘原子、卤素、氰基、羟基、氨基、硝基、C 1-6烷基、C 1-6卤代烷基、C 3-8环烷基或3-8元杂环基，其中所述的C 1-6烷基、C 1-6卤代烷基、氨基、C 3-8环烷基和3-8元杂环基任选进一步被选自氘原子、C 1-6烷基、C 1-6卤代烷基、卤素、羟基、氨基、硝基、氰基、C 1-6烷氧基、C 1-6羟烷基、C 3-8环烷基和3-6元杂环基中的一个或多个取代基所取代； R 3 and R 4 are the same or different, and are each independently selected from hydrogen atom, deuterium atom, halogen, cyano, hydroxyl, amino, nitro, C 1-6 alkyl, C 1-6 haloalkyl, C 3- 8 cycloalkyl or 3-8 membered heterocyclic group, wherein the C 1-6 alkyl, C 1-6 haloalkyl, amino, C 3-8 cycloalkyl and 3-8 membered heterocyclic group are optional Further selected from deuterium atom, C 1-6 alkyl, C 1-6 haloalkyl, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl and 3-6 membered heterocyclic group substituted by one or more substituents;
M为无机酸或有机酸，其中所述无机酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸或磷酸；有机酸选自2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4- 乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；M is an inorganic acid or an organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1- Hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-amino Benzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid , Lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, Galactonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid , Malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid Acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid ；
x选自0、1、2或3的整数；x is selected from an integer of 0, 1, 2 or 3;
n为0、1、2或3的整数；且n is an integer of 0, 1, 2 or 3; and
t为0、1、2、3、4、5或6的整数。t is an integer of 0, 1, 2, 3, 4, 5, or 6.
在本发明的一个优选实施例方案中，所述通式(I)所示化合物的晶型，其结构如通式(II)所示：In a preferred embodiment of the present invention, the crystal form of the compound represented by general formula (I) has a structure as shown in general formula (II):
Figure PCTCN2020082042-appb-000003
Figure PCTCN2020082042-appb-000003
其中：among them:
W、R 1、R 2、M、x和t如通式(I)所定义。 W, R 1 , R 2 , M, x and t are as defined in general formula (I).
在本发明的一个优选实施例方案中，所述的通式(I)所示化合物的晶型，其化合物结构如下：In a preferred embodiment of the present invention, the crystal form of the compound represented by general formula (I) has the following compound structure:
Figure PCTCN2020082042-appb-000004
Figure PCTCN2020082042-appb-000004
Figure PCTCN2020082042-appb-000005
Figure PCTCN2020082042-appb-000005
在本发明的一个优选实施例方案中，通式(I)所示化合物的晶型，所述化合物结构如式(III)所示：In a preferred embodiment of the present invention, the crystal form of the compound represented by general formula (I), and the structure of the compound is represented by formula (III):
Figure PCTCN2020082042-appb-000006
Figure PCTCN2020082042-appb-000006
x、M如通式(I)定义。x and M are as defined in general formula (I).
在本发明的一个进一步优选实施例方案中，其为式(III)化合物的自由碱晶型。优选地，其为无水物，进一步还包含水，所述水为管道水或结晶水，优选地， 含有0.5-8个水分子；更优选的，水合物含有0.5-4个水分子；进一步优选含有0.5-2.5个水分子；更进一步优选含有1个水分子。例如含有0.5、1、1.5、2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、7、7.5或8个水。In a further preferred embodiment of the present invention, it is the free base crystal form of the compound of formula (III). Preferably, it is an anhydrate, and further contains water, and the water is pipeline water or crystal water, preferably containing 0.5-8 water molecules; more preferably, the hydrate contains 0.5-4 water molecules; further It preferably contains 0.5-2.5 water molecules; more preferably, it contains 1 water molecule. For example, it contains 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 or 8 water.
在本发明的一个进一步优选实施例方案中，式(III)化合物的晶型，其中x为1-3；更进一步优选1。In a further preferred embodiment of the present invention, the crystalline form of the compound of formula (III), wherein x is 1 to 3;
在本发明的一个进一步优选实施例方案中，式(III)化合物的晶型为水合物或无水物。In a further preferred embodiment of the present invention, the crystal form of the compound of formula (III) is a hydrate or an anhydrate.
在本发明的一个进一步优选实施例方案中，式(III)化合物的晶型中含有0.5-8个水分子；优选的，水合物含有0.5-4个水分子；进一步优选含有0.5-2.5个水分子；更进一步优选含有1个水分子。例如含有0.5、1、1.5、2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、7、7.5或8个水。In a further preferred embodiment of the present invention, the crystal form of the compound of formula (III) contains 0.5-8 water molecules; preferably, the hydrate contains 0.5-4 water molecules; further preferably, it contains 0.5-2.5 water molecules. Molecule; More preferably, it contains 1 water molecule. For example, it contains 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 or 8 water.
在本发明的一个优选实施例方案中，M选自马来酸、对苯甲磺酸、盐酸、硝酸、硫酸、水杨酸、甲磺酸、苯磺酸或1,5-萘二磺酸；且x选自0.5、1、1.5或2，优选1。In a preferred embodiment of the present invention, M is selected from maleic acid, p-toluenesulfonic acid, hydrochloric acid, nitric acid, sulfuric acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid or 1,5-naphthalenedisulfonic acid And x is selected from 0.5, 1, 1.5 or 2, preferably 1.
在本发明的一个优选实施例方案中，通式(I)所示化合物晶型，其化合物结构如式(IV)所示：In a preferred embodiment of the present invention, the crystal form of the compound represented by general formula (I), and its compound structure is represented by formula (IV):
Figure PCTCN2020082042-appb-000007
Figure PCTCN2020082042-appb-000007
x、M如通式(I)定义。x and M are as defined in general formula (I).
本发明另外的目的在于提供一种通式(Ia)所示的化合物，其结构如式(Ia)所示:Another object of the present invention is to provide a compound represented by general formula (Ia), the structure of which is as shown in formula (Ia):
Figure PCTCN2020082042-appb-000008
Figure PCTCN2020082042-appb-000008
其中：among them:
W选自N或CH；W is selected from N or CH;
R 1选自氢原子、卤素、氨基、硝基、羟基、氰基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6卤代烷氧基、C 1-6羟烷基、C 3-8环烷基、-(CH 2) nR 3、-(CH 2) nOR 3、-(CH 2) nC(O)R 3、-(CH 2) nNR 3R 4或-(CH 2) nC(O)NR 3R 4； R 1 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R 3 , -(CH 2 ) n OR 3 , -(CH 2 ) n C(O)R 3 , -(CH 2 ) n NR 3 R 4 or -(CH 2 ) n C(O)NR 3 R 4 ;
R 2选自氢原子、卤素、氨基、硝基、羟基、氰基、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卤代烷氧基、C 3-8环烷基或3-8元杂环基； R 2 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl or 3-8 membered heterocyclic group;
R 3和R 4相同或不同，且各自独立地选自氢原子、氘原子、卤素、氰基、羟基、氨基、硝基、C 1-6烷基、C 1-6卤代烷基、C 3-8环烷基或3-8元杂环基，其中所述的C 1-6烷基、C 1-6卤代烷基、氨基、C 3-8环烷基和3-8元杂环基任选进一步被选自氘原子、C 1-6烷基、C 1-6卤代烷基、卤素、羟基、氨基、硝基、氰基、C 1-6烷氧基、C 1-6羟烷基、C 3-8环烷基和3-6元杂环基中的一个或多个取代基所取代； R 3 and R 4 are the same or different, and are each independently selected from hydrogen atom, deuterium atom, halogen, cyano, hydroxyl, amino, nitro, C 1-6 alkyl, C 1-6 haloalkyl, C 3- 8 cycloalkyl or 3-8 membered heterocyclic group, wherein the C 1-6 alkyl, C 1-6 haloalkyl, amino, C 3-8 cycloalkyl and 3-8 membered heterocyclic group are optional Further selected from deuterium atom, C 1-6 alkyl, C 1-6 haloalkyl, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl and 3-6 membered heterocyclic group substituted by one or more substituents;
M为无机酸或有机酸，其中所述无机酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸或磷酸；有机酸选自2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；M is an inorganic acid or an organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1- Hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-amino Benzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid , Lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, Galactonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid , Malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid Acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid ；
y为1、2或3的整数；y is an integer of 1, 2 or 3;
n为0、1、2或3的整数；且n is an integer of 0, 1, 2 or 3; and
t为0、1、2、3、4、5或6的整数。t is an integer of 0, 1, 2, 3, 4, 5, or 6.
在本发明一个优选的实施方案中，所述通式(Ia)所示的化合物，其结构如式(IIa)所示：In a preferred embodiment of the present invention, the compound represented by general formula (Ia) has a structure as shown in formula (IIa):
Figure PCTCN2020082042-appb-000009
Figure PCTCN2020082042-appb-000009
R 1、R 2、W、M、t、x如通式(I)定义。 R 1 , R 2 , W, M, t, and x are as defined in general formula (I).
在本发明一个优选的实施方案中，所述的通式(Ia)所示的化合物，其中，化合物结构如下：In a preferred embodiment of the present invention, the compound represented by the general formula (Ia), wherein the compound structure is as follows:
Figure PCTCN2020082042-appb-000010
Figure PCTCN2020082042-appb-000010
Figure PCTCN2020082042-appb-000011
Figure PCTCN2020082042-appb-000011
Figure PCTCN2020082042-appb-000012
Figure PCTCN2020082042-appb-000012
在本发明一个优选的实施方案中，所述的通式(Ia)所示的化合物，其化合物结构如式(IIIa)所示：In a preferred embodiment of the present invention, the compound represented by the general formula (Ia) has a compound structure as shown in the formula (IIIa):
Figure PCTCN2020082042-appb-000013
Figure PCTCN2020082042-appb-000013
M、y如通式(Ia)定义。M and y are as defined in general formula (Ia).
在本发明进一步优选的实施方案中，所述的通式(IIIa)所示的化合物为水无水物，进一步还包含水，所述水为管道水或结晶水。In a further preferred embodiment of the present invention, the compound represented by the general formula (IIIa) is a water anhydride, further comprising water, and the water is pipeline water or crystal water.
在本发明进一步优选的实施方案中，所述的通式(IIIa)所示的化合物中含有0.5-8个水分子；优选0.5-4个水分子；进一步优选0.5-2.5个水分子；更进一步优选1个水分子；例如含有0.5、1、1.5、2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、7、7.5或8个水。In a further preferred embodiment of the present invention, the compound represented by the general formula (IIIa) contains 0.5-8 water molecules; preferably 0.5-4 water molecules; further preferably 0.5-2.5 water molecules; Preferably 1 water molecule; for example, it contains 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 or 8 water.
在本发明进一步优选的实施方案中，所述的通式(Ia)所示的化合物，其化合物结构如式(IVa)所示：In a further preferred embodiment of the present invention, the compound represented by the general formula (Ia) has a compound structure as shown in formula (IVa):
Figure PCTCN2020082042-appb-000014
Figure PCTCN2020082042-appb-000014
M、y如通式(Ia)定义，优选地，M选自马来酸，y为1。M and y are as defined in general formula (Ia), preferably, M is selected from maleic acid, and y is 1.
本发明的目的还在于提供一种通式(I)所示化合物晶型或通式(Ia)所示化合物的制备方法，具体包括如下步骤：The object of the present invention is also to provide a method for preparing the crystal form of the compound represented by general formula (I) or the compound represented by general formula (Ia), which specifically includes the following steps:
1)称取适量的自由碱，用良性溶剂溶解；1) Weigh an appropriate amount of free base and dissolve it with a benign solvent;
2)任选地，称取适量的反离子酸，用有机溶剂溶解；优选地，反离子酸的量为1.2当量；2) Optionally, weigh an appropriate amount of counter ion acid and dissolve it with an organic solvent; preferably, the amount of counter ion acid is 1.2 equivalents;
3)任选地，把上述两种溶液合并，搅拌后若无沉淀析出，滴加不良溶剂直至出现浑浊，搅拌、析晶得目标产物；3) Optionally, combine the above two solutions, if there is no precipitation after stirring, add dropwise poor solvent until turbidity appears, stir and crystallize to obtain the target product;
其中：among them:
所述的良性溶剂选自甲醇、四氢呋喃、N,N-二甲基甲酰胺、N，N-二甲基乙酰胺或N-甲基吡咯烷酮；优选N,N-二甲基甲酰胺和N-甲基吡咯烷酮；The benign solvent is selected from methanol, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidone; preferably N,N-dimethylformamide and N- Methylpyrrolidone;
所述的有机溶剂选自甲醇、乙醇、乙酸乙酯、二氯甲烷、丙酮、正己烷、石油醚、苯、甲苯、氯仿、乙腈、四氯化碳、二氯乙烷、四氢呋喃、2-丁酮、3-戊酮、庚烷、甲基叔丁基醚、异丙醚、1,4-二氧六环、叔丁醇或N，N-二甲基甲酰胺；优选甲醇和乙醇；上述良性溶剂和有机溶液使用时需互溶；The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol and ethanol; the above The benign solvent and organic solution must be miscible when used;
所述的不良溶剂选自庚烷、水、甲基叔丁基醚、甲苯、异丙醚、乙酸乙酯、丙酮或乙腈；优选乙酸乙酯、甲基叔丁基醚、异丙醚和乙腈；The poor solvent is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably ethyl acetate, methyl tert-butyl ether, isopropyl ether and acetonitrile ；
所述的反离子酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸、磷酸、2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；优选甲磺酸、硫酸、盐酸、硝酸、苯磺酸、马来酸、己二酸、对甲基苯磺酸、柠檬酸、丙二酸和L-苹果酸；更优选马来酸。The counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroglutarate Amino acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid , 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphor acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid Acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably methanesulfonic acid, sulfuric acid, hydrochloric acid, nitric acid , Benzenesulfonic acid, maleic acid, adipic acid, p-toluenesulfonic acid, citric acid, malonic acid, and L-malic acid; maleic acid is more preferred.
本发明的目的还在于提供一种通式(I)所示化合物的晶型或通式(Ia)所示化合物的制备方法，具体包括如下步骤：The object of the present invention is also to provide a crystal form of the compound represented by general formula (I) or a method for preparing the compound represented by general formula (Ia), which specifically includes the following steps:
1)称取适量的自由碱，用不良性溶剂混悬；1) Weigh an appropriate amount of free base and suspend it in a poor solvent;
2)任选地，称取适量的反离子酸，用有机溶剂溶解；优选地，反离子酸的量为1.2当量；2) Optionally, weigh an appropriate amount of counter ion acid and dissolve it with an organic solvent; preferably, the amount of counter ion acid is 1.2 equivalents;
3)任选地，将2)中溶液加入1)中混悬液中，搅拌，析晶得目标产物；3) Optionally, add the solution in 2) to the suspension in 1), stir, and crystallize to obtain the target product;
其中：among them:
所述的不良性溶剂选自丙酮、乙酸乙酯、乙腈、乙醇、88％丙酮、四氢呋喃、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、叔丁醇或2-丁酮；优选甲醇、乙醇、四氢呋喃、乙酸乙酯、乙腈或丙酮；优选甲醇、乙醇、四氢呋喃、乙酸乙酯、乙腈和丙酮；The poor solvent is selected from acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N , N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol or 2-butanone; preferably methanol, ethanol, tetrahydrofuran, ethyl acetate, acetonitrile or acetone; preferably methanol, Ethanol, tetrahydrofuran, ethyl acetate, acetonitrile and acetone;
所述的有机溶剂选自甲醇、乙醇、乙酸乙酯、二氯甲烷、丙酮、正己烷、石油醚、苯、甲苯、氯仿、乙腈、四氯化碳、二氯乙烷、四氢呋喃、2-丁酮、3-戊酮、庚烷、甲基叔丁基醚、异丙醚、1,4-二氧六环、叔丁醇或N，N-二甲基甲酰胺；优选甲醇或乙醇；上述良性溶剂和有机溶液使用时需互溶；The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol or ethanol; the above The benign solvent and organic solution must be miscible when used;
所述的反离子酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸、磷酸、2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；优选马来酸。The counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroglutarate Amino acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid , 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphor acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid Acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably maleic acid.
本发明的目的还在于提供一种通式(I)所示化合物的晶型或通式(Ia)所示化合物的制备方法，具体包括如下步骤：The object of the present invention is also to provide a crystal form of the compound represented by general formula (I) or a method for preparing the compound represented by general formula (Ia), which specifically includes the following steps:
1)称取适量的自由碱，加入良性溶剂，加热至溶解；1) Weigh an appropriate amount of free base, add a benign solvent, and heat to dissolve;
2)任选地，称取适量的反离子酸，用有机溶剂溶解；优选地，反离子酸的量1.2当量；2) Optionally, weigh an appropriate amount of counter ion acid and dissolve it with an organic solvent; preferably, the amount of counter ion acid is 1.2 equivalents;
3)搅拌、降温析晶得到目标产物；3) Stirring, cooling and crystallization to obtain the target product;
其中：among them:
所述的良性溶剂选自甲醇、四氢呋喃、N,N-二甲基甲酰胺、N，N-二甲基乙酰胺或N-甲基吡咯烷酮；优选N,N-二甲基甲酰胺和N-甲基吡咯烷酮；The benign solvent is selected from methanol, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidone; preferably N,N-dimethylformamide and N- Methylpyrrolidone;
所述的有机溶剂选自甲醇、乙醇、乙酸乙酯、二氯甲烷、丙酮、正己烷、石油醚、苯、甲苯、氯仿、乙腈、四氯化碳、二氯乙烷、四氢呋喃、2-丁酮、3-戊酮、庚烷、甲基叔丁基醚、异丙醚、1,4-二氧六环、叔丁醇或N，N-二甲基甲酰胺；优选甲醇和乙醇；上述良性溶剂和有机溶液使用时需互溶；The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol and ethanol; the above The benign solvent and organic solution must be miscible when used;
所述的反离子酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸、磷酸、2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、 己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；优选甲磺酸、硫酸、盐酸、硝酸、苯磺酸、马来酸、己二酸、对甲基苯磺酸、柠檬酸、丙二酸或L-苹果酸；更优选马来酸。The counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyrogallium Amino acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid , 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphor acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid Acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably methanesulfonic acid, sulfuric acid, hydrochloric acid, nitric acid , Benzenesulfonic acid, maleic acid, adipic acid, p-toluenesulfonic acid, citric acid, malonic acid or L-malic acid; more preferably maleic acid.
在本发明的一个优选实施方案中，所述的通式(IV)所示化合物的晶型，其中M选自马来酸、对苯甲磺酸、盐酸、硝酸、水杨酸、甲磺酸、苯磺酸或1,5-萘二磺酸；优选马来酸，进一步优选x为1。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is selected from maleic acid, p-toluenesulfonic acid, hydrochloric acid, nitric acid, salicylic acid, methanesulfonic acid , Benzenesulfonic acid or 1,5-naphthalenedisulfonic acid; preferably maleic acid, more preferably x is 1.
在本发明的一个优选实施方案中，所述的通式(IV)所示化合物的晶型，其自由碱结晶形式I，X-射线粉末衍射图谱在在2θ为6.4和26.7处(2θ±0.2°)具有特征峰，优选地，还包含在2θ为7.2、13.1、16.8、17.7、18.9、20.2、21.2和28.2处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystalline form of the compound represented by the general formula (IV), the free base crystalline form I, the X-ray powder diffraction pattern is at 2θ of 6.4 and 26.7 (2θ±0.2 °) has characteristic peaks, preferably, it further includes characteristic peaks at 2θ of 7.2, 13.1, 16.8, 17.7, 18.9, 20.2, 21.2, and 28.2 (2θ±0.2°).
本发明所述的通式(IV)所示化合物的晶型，其为自由碱化合物晶型，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。The crystal form of the compound represented by the general formula (IV) of the present invention, which is a free base compound crystal form, uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d are shown in the table 1 shown.
表1Table 1
Figure PCTCN2020082042-appb-000015
Figure PCTCN2020082042-appb-000015
Figure PCTCN2020082042-appb-000016
Figure PCTCN2020082042-appb-000016
本发明所述通式(IV)所示化合物的晶型，其为自由碱化合物的晶型I，其X-射线粉末衍射图谱基本如图1所示。The crystalline form of the compound represented by the general formula (IV) of the present invention is the crystalline form I of the free base compound, and its X-ray powder diffraction pattern is basically shown in FIG. 1.
本发明所述通式(IV)所示化合物的晶型，其为自由碱化合物的晶型I，其TGA图谱基本如图2所示。The crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form I of the free base compound, and its TGA pattern is basically shown in FIG. 2.
本发明所述通式(IV)所示化合物的晶型，其为自由碱化合物的晶型I，其DSC图谱基本如图3所示。The crystalline form of the compound represented by the general formula (IV) of the present invention is the crystalline form I of the free base compound, and its DSC spectrum is basically shown in FIG. 3.
在本发明的一个优选实施例方案中，所述的通式(IV)所示化合物的晶型，其自由碱结晶形式II，X-射线粉末衍射图谱在2θ为8.5、13.4和17.0处(2θ±0.2°)具有特征峰，还包含在2θ为10.2、10.9、12.8、13.1、16.1、17.8、18.8、19.5、23.0、23.9、24.4、25.5、26.1和27.6处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystalline form of the compound represented by the general formula (IV), its free base crystalline form II, the X-ray powder diffraction pattern is at 2θ of 8.5, 13.4 and 17.0 (2θ ±0.2°) has characteristic peaks, and also includes characteristic peaks at 2θ of 10.2, 10.9, 12.8, 13.1, 16.1, 17.8, 18.8, 19.5, 23.0, 23.9, 24.4, 25.5, 26.1 and 27.6 (2θ±0.2°) .
本发明所述的通式(IV)所示化合物的晶型，其为自由碱化合物晶型，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。The crystal form of the compound represented by the general formula (IV) of the present invention, which is a free base compound crystal form, uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks expressed in 2θ angles and interplanar spacing d are shown in the table 2 shown.
表2Table 2
Figure PCTCN2020082042-appb-000017
Figure PCTCN2020082042-appb-000017
Figure PCTCN2020082042-appb-000018
Figure PCTCN2020082042-appb-000018
本发明所述通式(IV)所示化合物的晶型，其为自由碱化合物的晶型II，其X-射线粉末衍射图谱基本如图4所示。The crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form II of the free base compound, and its X-ray powder diffraction pattern is basically as shown in FIG. 4.
本发明所述通式(IV)所示化合物的晶型，其为自由碱化合物的晶型II，其 TGA图谱基本如图5所示。The crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form II of the free base compound, and its TGA pattern is basically shown in FIG. 5.
本发明所述通式(IV)所示化合物的晶型，其为自由碱化合物的晶型II，其DSC图谱基本如图6所示。The crystal form of the compound represented by the general formula (IV) of the present invention is the crystal form II of the free base compound, and its DSC spectrum is basically as shown in FIG. 6.
在本发明的一个优选实施例方案中，所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型I，X-射线粉末衍射图谱在2θ为5.5和16.3处(2θ±0.2°)具有特征峰；进一步还包含在2θ为10.8、15.3、17.7、26.1、26.4和27.0处(2θ±0.2°)具有特征峰；更进一步在2θ为13.0、14.9、20.0和20.8处(2θ±0.2°)具有特征峰；In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, the maleate crystal form I, X-ray The powder diffraction pattern has characteristic peaks at 2θ of 5.5 and 16.3 (2θ±0.2°); it further includes characteristic peaks at 2θ of 10.8, 15.3, 17.7, 26.1, 26.4 and 27.0 (2θ±0.2°); further There are characteristic peaks at 2θ of 13.0, 14.9, 20.0 and 20.8 (2θ±0.2°);
本发明所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型I，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。The crystal form of the compound represented by the general formula (IV) of the present invention, wherein M is maleic acid, and x is 1, and the maleate salt crystal form I, using Cu-Kα radiation, is set at a 2θ angle and a crystal plane The X-ray characteristic diffraction peaks represented by the spacing d are shown in Table 3.
表3table 3
Figure PCTCN2020082042-appb-000019
Figure PCTCN2020082042-appb-000019
Figure PCTCN2020082042-appb-000020
Figure PCTCN2020082042-appb-000020
在本发明的一个优选实施例方案中，所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型I，其X-射线粉末衍射图谱基本如图7所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form I, and its X- The X-ray powder diffraction pattern is basically shown in Figure 7.
在本发明的一个优选实施例方案中，所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型I，其TGA图谱基本如图8所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form I, and its TGA pattern Basically as shown in Figure 8.
在本发明的一个优选实施例方案中，所述的所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型I，其DSC图谱基本如图9所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, the maleate salt crystal form I, The DSC spectrum is basically shown in Figure 9.
在本发明的一个优选实施例方案中，所述的所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型II，X-射线粉末衍射图谱在2θ为5.5、10.8、16.3和17.6处(2θ±0.2°)具有特征峰；进一步还包含在2θ为14.0、26.4、26.6和27.2的特征峰；更进一步包含在2θ为8.8、12.9、18.4、18.9、20.6、21.5、22.4和22.8处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form II, The X-ray powder diffraction pattern has characteristic peaks at 2θ of 5.5, 10.8, 16.3 and 17.6 (2θ±0.2°); it further contains characteristic peaks at 2θ of 14.0, 26.4, 26.6 and 27.2; There are characteristic peaks at 8.8, 12.9, 18.4, 18.9, 20.6, 21.5, 22.4 and 22.8 (2θ±0.2°).
本发明所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型II，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。The crystalline form of the compound represented by the general formula (IV) of the present invention, wherein M is maleic acid, and x is 1, and its maleate salt crystalline form II uses Cu-Kα radiation, with a 2θ angle and crystal plane The X-ray characteristic diffraction peaks represented by the spacing d are shown in Table 4.
表4Table 4
Figure PCTCN2020082042-appb-000021
Figure PCTCN2020082042-appb-000021
Figure PCTCN2020082042-appb-000022
Figure PCTCN2020082042-appb-000022
在本发明的一个优选实施例方案中，所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型II，其X-射线粉末衍射图谱基本如图10所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form II, and its X- The X-ray powder diffraction pattern is basically shown in Figure 10.
在本发明的一个优选实施例方案中，所述的通式(IV)所示化合物的晶型， 其中M为马来酸，且x为1，其马来酸盐晶型II，其DSC图谱基本如图11所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form II, and its DSC spectrum Basically as shown in Figure 11.
在本发明的一个优选实施例方案中，所述的通式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型III，X-射线粉末衍射图谱在2θ为4.9、15.1、17.2、17.5、26.5和26.9处(2θ±0.2°)具有特征峰；进一步还包含在2θ为9.8、14.7、18.4和19.8处(2θ±0.2°)具有特征峰；更进一步包含在2θ为12.3、13.0、14.0和27.5处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by the general formula (IV), wherein M is maleic acid, and x is 1, the maleate crystal form III, X-ray The powder diffraction pattern has characteristic peaks at 2θ of 4.9, 15.1, 17.2, 17.5, 26.5 and 26.9 (2θ±0.2°); it further contains characteristic peaks at 2θ of 9.8, 14.7, 18.4 and 19.8 (2θ±0.2°) Peak; further includes characteristic peaks at 2θ of 12.3, 13.0, 14.0, and 27.5 (2θ±0.2°).
本发明所述的通式(IV)所示化合物，其中M为马来酸，且x为1，其马来酸盐晶型III，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5所示。The compound represented by the general formula (IV) of the present invention, wherein M is maleic acid and x is 1, its maleate crystal form III, using Cu-Kα radiation, with 2θ angle and interplanar spacing d value The characteristic X-ray diffraction peaks are shown in Table 5.
表5table 5
Figure PCTCN2020082042-appb-000023
Figure PCTCN2020082042-appb-000023
Figure PCTCN2020082042-appb-000024
Figure PCTCN2020082042-appb-000024
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型III，其X-射线粉末衍射图谱基本如图12所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form III, and its X-ray The powder diffraction pattern is basically shown in Figure 12.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型III，其DSC图谱基本如图13所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is maleic acid, and x is 1, its maleate salt crystal form III, and its DSC spectrum is basically As shown in Figure 13.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物晶型，其中M为马来酸，且x为1，其马来酸盐晶型IV，X-射线粉末衍射图谱在2θ为具有4.8、16.7的特征峰；进一步还包含在2θ为9.6、11.9和14.9处(2θ±0.2°)具有特征峰；更进一步包含在2θ为7.2、12.9、14.3、19.1、19.3、21.5和23.9处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form IV, X-ray powder diffraction The spectrum has characteristic peaks of 4.8 and 16.7 in 2θ; it further includes characteristic peaks at 2θ of 9.6, 11.9 and 14.9 (2θ±0.2°); it is further included in 2θ of 7.2, 12.9, 14.3, 19.1, 19.3, There are characteristic peaks at 21.5 and 23.9 (2θ±0.2°).
本发明所述的式(IV)所示化合物，其中M为马来酸，且x为1，其马来酸盐晶型IV，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6所示。The compound represented by formula (IV) of the present invention, wherein M is maleic acid and x is 1, its maleate crystal form IV, using Cu-Kα radiation, is expressed by 2θ angle and interplanar spacing d value The characteristic X-ray diffraction peaks are shown in Table 6.
表6Table 6
Figure PCTCN2020082042-appb-000025
Figure PCTCN2020082042-appb-000025
Figure PCTCN2020082042-appb-000026
Figure PCTCN2020082042-appb-000026
Figure PCTCN2020082042-appb-000027
Figure PCTCN2020082042-appb-000027
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型IV，其X-射线粉末衍射图谱基本如图14所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form IV, and its X-ray The powder diffraction pattern is basically shown in Figure 14.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为马来酸，且x为1，其马来酸盐晶型IV，其DSC图谱基本如图15所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is maleic acid, and x is 1, its maleate crystal form IV, and its DSC spectrum is basically As shown in Figure 15.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为盐酸，其盐酸盐晶型，X-射线粉末衍射图谱在2θ为5.8和17.2处(2θ±0.2°)具有特征峰；进一步还包含在2θ为12.0、13.6、16.4、21.7、23.0、26.1、26.3和27.1处(2θ±0.2°)具有特征峰；更进一步包含在2θ为8.9、28.8和30.1处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is hydrochloric acid, the hydrochloride crystal form, and the X-ray powder diffraction pattern is at 2θ of 5.8 and 17.2 (2θ±0.2°) has characteristic peaks; further includes characteristic peaks at 2θ of 12.0, 13.6, 16.4, 21.7, 23.0, 26.1, 26.3, and 27.1 (2θ±0.2°); it is further included in 2θ of 8.9, There are characteristic peaks at 28.8 and 30.1 (2θ±0.2°).
本发明所述的式(IV)所示化合物，其中M为盐酸，其盐酸盐晶型，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7所示。The compound represented by the formula (IV) of the present invention, wherein M is hydrochloric acid, its hydrochloride crystal form, using Cu-Kα radiation, the X-ray characteristic diffraction peaks expressed in 2θ angle and interplanar spacing d are shown in the table 7 shown.
表7Table 7
Figure PCTCN2020082042-appb-000028
Figure PCTCN2020082042-appb-000028
Figure PCTCN2020082042-appb-000029
Figure PCTCN2020082042-appb-000029
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为盐酸，其盐酸盐晶型，其X-射线粉末衍射图谱基本如图16所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is hydrochloric acid, its hydrochloride crystal form, and its X-ray powder diffraction pattern is basically as shown in Figure 16. .
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型I，X-射线粉末衍射图谱在2θ为6.1、11.3、17.0、17.5和18.3处(2θ±0.2°)具有特征峰；进一步还包含在2θ为12.7、19.1、19.9、20.6和22.2处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form I, X-ray powder diffraction The spectrum has characteristic peaks at 2θ of 6.1, 11.3, 17.0, 17.5 and 18.3 (2θ±0.2°); it further contains characteristic peaks at 2θ of 12.7, 19.1, 19.9, 20.6 and 22.2 (2θ±0.2°).
本发明所述的式(IV)所示化合物，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型I，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8所示。The compound represented by formula (IV) of the present invention, wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form I, using Cu-Kα radiation, expressed in 2θ angle and interplanar spacing d value The characteristic X-ray diffraction peaks are shown in Table 8.
表8Table 8
Figure PCTCN2020082042-appb-000030
Figure PCTCN2020082042-appb-000030
Figure PCTCN2020082042-appb-000031
Figure PCTCN2020082042-appb-000031
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型I，其X-射线粉末衍射图谱基本如图17所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form I, and its X-ray powder The diffraction pattern is basically shown in Figure 17.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型II，X-射线粉末衍射图谱在2θ为5.4、8.6、11.4、16.8、18.2、19.7、20.4和21.9处(2θ±0.2°)具有特征峰；进一步还包含在2θ为13.5、13.7、15.7、17.2、23.7、25.5和27.6处(2θ±0.2°)具有特征峰；更进一步包含在2θ为16.0、21.4、25.9和28.7处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form II, X-ray powder diffraction The spectrum has characteristic peaks at 2θ of 5.4, 8.6, 11.4, 16.8, 18.2, 19.7, 20.4, and 21.9 (2θ±0.2°); it is further included at 2θ of 13.5, 13.7, 15.7, 17.2, 23.7, 25.5 and 27.6 (2θ±0.2°) has characteristic peaks; and further includes characteristic peaks at 2θ of 16.0, 21.4, 25.9, and 28.7 (2θ±0.2°).
本发明所述的式(IV)所示化合物，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型II，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9所示。The compound represented by formula (IV) of the present invention, wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form II, using Cu-Kα radiation, expressed in 2θ angle and interplanar spacing d value The characteristic X-ray diffraction peaks are shown in Table 9.
表9Table 9
Figure PCTCN2020082042-appb-000032
Figure PCTCN2020082042-appb-000032
Figure PCTCN2020082042-appb-000033
Figure PCTCN2020082042-appb-000033
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型II，其X-射线粉末衍射图谱基本如图18所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form II, and its X-ray powder The diffraction pattern is basically shown in Figure 18.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其 中M为对苯甲磺酸，其对苯甲磺酸盐晶型III，X-射线粉末衍射图谱在2θ为4.9、8.6、13.2、18.9、20.6和25.2处(2θ±0.2°)具有特征峰；进一步还包含在2θ为9.6、10.9、12.6、15.0、15.6、17.0、22.6、25.8和27.5处(2θ±0.2°)具有特征峰；更进一步包含在2θ为10.5、13.9、16.5、17.7、21.7、26.1、26.6、26.9、27.8、29.8和32.2处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form III, X-ray powder diffraction The spectrum has characteristic peaks at 2θ of 4.9, 8.6, 13.2, 18.9, 20.6, and 25.2 (2θ±0.2°); it is further included at 2θ of 9.6, 10.9, 12.6, 15.0, 15.6, 17.0, 22.6, 25.8, and 27.5. (2θ±0.2°) has characteristic peaks; and further includes characteristic peaks at 2θ of 10.5, 13.9, 16.5, 17.7, 21.7, 26.1, 26.6, 26.9, 27.8, 29.8, and 32.2 (2θ±0.2°).
本发明所述的式(IV)所示化合物，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型III，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10所示。The compound represented by formula (IV) of the present invention, wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form III, using Cu-Kα radiation, expressed in 2θ angle and interplanar spacing d value X-ray characteristic diffraction peaks are shown in Table 10.
表10Table 10
Figure PCTCN2020082042-appb-000034
Figure PCTCN2020082042-appb-000034
Figure PCTCN2020082042-appb-000035
Figure PCTCN2020082042-appb-000035
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型III，其X-射线粉末衍射图谱基本如图19所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form III, and its X-ray powder The diffraction pattern is basically shown in Figure 19.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型IV，X-射线粉末衍射图谱在2θ为5.5、11.9、16.3、19.4和25.5处(2θ±0.2°)具有特征峰；进一步还包含在2θ为13.8、18.1、18.6、20.1、21.3、22.9和26.5处(2θ±0.2°)具有特征峰；更进一步包含8.7、9.1、10.2、10.8、12.9和20.4处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form IV, X-ray powder diffraction The spectrum has characteristic peaks at 2θ of 5.5, 11.9, 16.3, 19.4, and 25.5 (2θ±0.2°); it further includes 2θ at 13.8, 18.1, 18.6, 20.1, 21.3, 22.9, and 26.5 (2θ±0.2°) It has characteristic peaks; it further includes characteristic peaks at 8.7, 9.1, 10.2, 10.8, 12.9 and 20.4 (2θ±0.2°).
本发明所述的式(IV)所示化合物，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型IV，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11所示。The compound represented by formula (IV) of the present invention, wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form IV, using Cu-Kα radiation, expressed by 2θ angle and interplanar spacing d value The characteristic X-ray diffraction peaks are shown in Table 11.
表11Table 11
Figure PCTCN2020082042-appb-000036
Figure PCTCN2020082042-appb-000036
Figure PCTCN2020082042-appb-000037
Figure PCTCN2020082042-appb-000037
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为对苯甲磺酸，其对苯甲磺酸盐晶型IV，其X-射线粉末衍射图谱基本如图20所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is p-toluenesulfonic acid, its p-toluenesulfonic acid salt crystal form IV, and its X-ray powder The diffraction pattern is basically shown in Figure 20.
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为硝酸，其硝酸盐晶型，X-射线粉末衍射图谱在2θ为5.0、16.3、16.7和28.0处(2θ±0.2°)具有特征峰；进一步还包含在2θ为8.2、8.5、11.8、13.2和29.1处(2θ±0.2°)具有特征峰；更进一步包含在2θ为19.7、20.4、21.0和24.5处(2θ±0.2°)具有特征峰。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is nitric acid, its nitrate crystal form, and the X-ray powder diffraction pattern at 2θ is 5.0, 16.3, 16.7 And 28.0 (2θ±0.2°) with characteristic peaks; further include characteristic peaks at 2θ of 8.2, 8.5, 11.8, 13.2, and 29.1 (2θ±0.2°); further including 2θ at 19.7, 20.4, 21.0 And 24.5 (2θ±0.2°) have characteristic peaks.
本发明所述的式(IV)所示化合物，其中M为硝酸，其硝酸盐晶型，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12所示。The compound represented by the formula (IV) of the present invention, wherein M is nitric acid, and its nitrate crystal form uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks expressed by 2θ angle and interplanar spacing d are shown in Table 12. Shown.
表12Table 12
Figure PCTCN2020082042-appb-000038
Figure PCTCN2020082042-appb-000038
Figure PCTCN2020082042-appb-000039
Figure PCTCN2020082042-appb-000039
在本发明的一个优选实施例方案中，所述的式(IV)所示化合物的晶型，其中M为硝酸，其硝酸盐晶型，其X-射线粉末衍射图谱基本如图21所示。In a preferred embodiment of the present invention, the crystal form of the compound represented by formula (IV), wherein M is nitric acid, its nitrate crystal form, and its X-ray powder diffraction pattern is basically as shown in FIG. 21.
本发明的目的还在于提供了一种药物组合物，其含有治疗有效量的所述式(I)所示通式化合物的晶型或式(Ia)所示通式化合物，以及一种或多种药学上可接受的载体。The object of the present invention is also to provide a pharmaceutical composition, which contains a therapeutically effective amount of the crystal form of the compound of formula (I) or the compound of formula (Ia), and one or more A pharmaceutically acceptable carrier.
本发明的目的还在于提供了所述式(I)所示通式化合物晶型或式(Ia)所示通式化合物以及所述的药物组合物在制备用于治疗和/或预防由ERK介导的癌症或肿瘤相关疾病的药物中的用途。The purpose of the present invention is also to provide the crystal form of the general formula compound represented by formula (I) or the general formula compound represented by formula (Ia) and the pharmaceutical composition in the preparation for the treatment and/or prevention of ERK-mediated The use of drugs for leading cancer or tumor-related diseases.
本发明的目的还在于提供了所述式(I)所示通式化合物的晶型或式(Ia)所示通式化合物及其药物组合物在制备治疗癌症、炎症、慢性肝病、糖尿病、心血管疾病或AIDS的药物中的应用。The object of the present invention is also to provide the crystal form of the compound represented by formula (I) or the compound represented by formula (Ia) and its pharmaceutical composition for preparing and treating cancer, inflammation, chronic liver disease, diabetes, heart disease Application in medicine for vascular disease or AIDS.
本发明的目的还在于提供了一种治疗预防和/或治疗预防ERK介导的病理学特征的疾病的方法，其包括向患者施用治疗有效剂量的式(I)所示通式化合物的晶体或式(Ia)所示通式化合物及其药物组合物。其中ERK介导的病理学特征的疾病包括癌症、炎症、慢性肝病、糖尿病、心血管疾病或AIDS的疾病。The object of the present invention is also to provide a method for the treatment, prevention and/or treatment and prevention of ERK-mediated pathological characteristics of diseases, which comprises administering to a patient a therapeutically effective dose of a crystal of a compound of formula (I) or Compounds of general formula represented by formula (Ia) and pharmaceutical compositions thereof. Diseases with ERK-mediated pathological characteristics include cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease or AIDS.
以上所述的用途，其中所述的疾病选自癌症、骨病、炎性疾病、免疫疾病、神经系统疾病、代谢性疾病、呼吸性疾病或心脏病中的药物应用；其中所述癌症选自乳腺癌、胰腺癌、非小细胞肺癌、甲状腺癌、精原细胞瘤、黑素瘤、膀胱癌、肝癌、肾癌、骨髓增生异常综合征、急性髓性白血病或结直肠癌。The above-mentioned use, wherein the disease is selected from cancer, bone disease, inflammatory disease, immune disease, neurological disease, metabolic disease, respiratory disease, or medical application in heart disease; wherein the cancer is selected from Breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, acute myeloid leukemia, or colorectal cancer.
发明的详细说明Detailed description of the invention
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。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 The alkyl group is 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-ethyl Hexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethyl Base hexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl and various branched isomers. Alkyl groups may be substituted or unsubstituted. When substituted, the substituents may be substituted at any available attachment point. The substituents are preferably one or more of the following groups, which are independently selected from alkanes Group, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkane Oxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate group, the present invention preferably methyl, ethyl, isopropyl, tert-butyl, haloalkyl , 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，上述取代基可以连接在不同的碳原子形成碳链，也可以连接在一个碳原子上形成环烷基。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" It refers to -(CH 2 ) 3 -, "butylene" refers to -(CH 2 ) 4 -, etc. The above-mentioned substituents can be connected to different carbon atoms to form a carbon chain, or they can be connected to a carbon atom to form a cycloalkyl group. The term "alkenyl" refers to an alkyl group as defined above composed of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3 -Butenyl etc. Alkenyl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from 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8个碳原子，更优选包含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、环己基、环戊基和环庚基。The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent. The cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 8 carbon atoms, more preferably 3 to 6 Carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene Polycyclic cycloalkyl groups include spiro, condensed and bridged cycloalkyl groups, preferably cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl and cycloheptyl.
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8个环原子；最优选包含3至8个环原子。单环杂环基的非限制性实例包括吡咯烷基、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、哌嗪基、吗啉基、硫代吗啉基、高哌嗪基、吡喃基等，优选四氢呋喃基、吡唑烷基、吗啉基、哌嗪基和吡喃基。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；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。 The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent which contains 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (where m is an integer of 0 to 2) heteroatoms, but does not include the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably contains 3 to 8 ring atoms; most preferably contains 3 to 8 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidine Group, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably tetrahydrofuryl, pyrazolidinyl, morpholinyl, piperazinyl and pyranyl. Polycyclic heterocyclic groups include spiro, condensed and bridged heterocyclic groups; the spiro, condensed and bridged heterocyclic groups are optionally connected to other groups through a single bond, or through a ring Any two or more of the above atoms are further connected to other cycloalkyl groups, heterocyclic groups, aryl groups and heteroaryl groups; heterocyclic groups may be optionally substituted or unsubstituted, when substituted , The substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano , Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy, 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8个环原子；最优选包含3至8个环原子。单环杂环基的非限制性实例包括氧杂环丁烷基、吡咯烷基、吡咯烷酮基、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、哌嗪基、吗啉基、硫代吗啉基、高哌嗪基、吡喃基等，优选氧杂环丁烷基、吡咯烷酮基、四氢呋喃基、吡唑烷基、吗啉基、哌嗪基和吡喃基。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 which contains 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (where m is an integer of 0 to 2) heteroatoms, but does not include the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably contains 3 to 8 ring atoms; most preferably contains 3 to 8 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include oxetanyl, pyrrolidinyl, pyrrolidone, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrrolidinyl Azolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably oxetanyl, pyrrolidonyl, tetrahydrofuranyl, Pyrazolidinyl, morpholinyl, piperazinyl and pyranyl. Polycyclic heterocyclic groups include spiro, condensed and bridged heterocyclic groups; the spiro, condensed and bridged heterocyclic groups are optionally connected to other groups through a single bond, or through a ring Any two or more of the above atoms are further connected to other cycloalkyl groups, heterocyclic groups, aryl groups and heteroaryl groups.
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个碳原子的烷氧基，更优选1至6个碳原子的烷氧基，最更优选1至3个碳原子的烷氧基。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), where alkyl is defined as described above. It is preferably an alkoxy group having 1 to 8 carbon atoms, more preferably an alkoxy group having 1 to 6 carbon atoms, and most preferably an alkoxy group having 1 to 3 carbon atoms. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. The alkoxy group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkane 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" refers to an alkyl group substituted with one or more halogens, where 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" refers to an alkoxy group substituted with one or more halogens, where alkoxy 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" refers to an alkyl group substituted with a hydroxy group, where the alkyl group is as defined above.
“烯基”指链烯基，又称烯烃基，优选含有2至8个碳原子的烯基，更优选2至6个碳原子的烯基，最更优选2至3个碳原子的烯基；其中所述的烯基可以进 一步被其他相关基团取代，例如：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。"Alkenyl" refers to an alkenyl group, also known as an alkene group, preferably an alkenyl group containing 2 to 8 carbon atoms, more preferably an alkenyl group having 2 to 6 carbon atoms, and most preferably an alkenyl group having 2 to 3 carbon atoms ; The alkenyl group can be further substituted by other related groups, such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano , Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.
“炔基”指(CH≡C-)，优选含有2至8个碳原子的炔基，更优选2至6个碳原子的炔基，最更优选2至3个碳原子的炔基。其中所述的炔基可以进一步被其他相关基团取代，例如：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。"Alkynyl" refers to (CH≡C-), preferably an alkynyl group containing 2 to 8 carbon atoms, more preferably an alkynyl group having 2 to 6 carbon atoms, and most preferably an alkynyl group having 2 to 3 carbon atoms. The alkynyl group described therein may be further substituted by other related groups, such as: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy, or carboxylate.
“羟基”指-OH基团。"Hydroxy" refers to the -OH group.
“卤素”指氟、氯、溴或碘。"Halogen" refers to fluorine, chlorine, bromine or iodine.
“氨基”指-NH 2。 "Amino" refers to -NH 2 .
“氰基”指-CN。"Cyano" refers to -CN.
“硝基”指-NO 2。 "Nitro" refers to -NO 2 .
“羧基”指-C(O)OH。"Carboxy" refers to -C(O)OH.
“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。"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" and other terms all express the same Meaning, which means that X can be any one or more of A, B, and C.
“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。"Optional" or "optionally" means that the event or environment described later can but does not have to occur, and the description includes occasions where the event or environment occurs or does not occur.
“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" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of each other, substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without too much effort. For example, an amino group or a hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (eg, olefinic) bond.
“立体异构”包含几何异构(顺反异构)、旋光异构、构象异构三类。"Stereoisomerism" includes geometric isomerism (cis-trans isomerism), optical isomerism, and conformational isomerism.
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 of the present invention can also be replaced by a deuterium atom.
“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 of the compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, and other components such as physiological/pharmaceutically acceptable carriers And excipients. The purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredients and thus the biological activity.
“可药用盐”是指本发明化合物的盐，这类盐用于哺乳动物体内时具有安全性和有效性，且具有应有的生物活性。"Pharmaceutically acceptable salt" refers to the salt of the compound of the present invention. Such salt is safe and effective when used in mammals, and has due biological activity.
“TGA”是指热重分析(TGA)实验。"TGA" refers to thermogravimetric analysis (TGA) experiments.
“DSC”是指差示扫描量热法(DSC)实验。"DSC" refers to differential scanning calorimetry (DSC) experiments.
“DVS”是指动态水分吸附(DVS)实验。"DVS" refers to dynamic moisture adsorption (DVS) experiments.
“XRPD”是指X-射线粉末衍射(XRPD)实验。"XRPD" refers to X-ray powder diffraction (XRPD) experiments.
“HPLC”是指高效液相色谱(HPLC)实验。"HPLC" refers to high performance liquid chromatography (HPLC) experiments.
“PK”是指药物代谢动力学(PK)实验。"PK" refers to pharmacokinetic (PK) experiments.
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.
附图说明Description of the drawings
图1为自由碱晶型I的XRPD图示。Figure 1 is an XRPD diagram of the free base crystal form I.
图2为自由碱晶型I的TGA图示。Figure 2 is a TGA diagram of the free base crystal form I.
图3为自由碱晶型I的DSC图示。Figure 3 is a DSC diagram of the free base crystal form I.
图4为自由碱晶型II的XRPD图示。Figure 4 is an XRPD diagram of the free base crystal form II.
图5为自由碱晶型II的TGA图示。Figure 5 is a TGA diagram of the free base crystal form II.
图6为自由碱晶型II的DSC图示。Figure 6 is a DSC chart of the free base crystal form II.
图7为马来酸盐晶型I的XRPD图示。Figure 7 is an XRPD diagram of maleate salt form I.
图8为马来酸盐晶型I的TGA图示。Figure 8 is a TGA diagram of maleate salt form I.
图9为马来酸盐晶型I的DSC图示。Figure 9 is a DSC diagram of maleate salt form I.
图10为马来酸盐晶型II的XRPD图示。Figure 10 is an XRPD diagram of maleate salt form II.
图11为马来酸盐晶型II的DSC图示。Figure 11 is a DSC chart of maleate salt form II.
图12为马来酸盐晶型III的XRPD图示。Figure 12 is an XRPD diagram of maleate salt form III.
图13为马来酸盐晶型III的DSC图示。Figure 13 is a DSC chart of maleate salt form III.
图14为马来酸盐晶型IV的XRPD图示。Figure 14 is an XRPD diagram of maleate salt form IV.
图15为马来酸盐晶型IV的DSC图示。Figure 15 is a DSC chart of maleate salt form IV.
图16为盐酸盐的XRPD图示。Figure 16 is an XRPD diagram of the hydrochloride salt.
图17为对苯甲磺酸盐晶型I的XRPD图示。Figure 17 is an XRPD diagram of p-toluenesulfonate form I.
图18为对苯甲磺酸盐晶型II的XRPD图示。Figure 18 is an XRPD diagram of p-toluenesulfonate Form II.
图19为对苯甲磺酸盐晶型III的XRPD图示。Figure 19 is an XRPD diagram of p-toluenesulfonate Form III.
图20为对苯甲磺酸盐晶型IV的XRPD图示。Figure 20 is an XRPD diagram of p-toluenesulfonate salt form IV.
图21为硝酸酸盐的XRPD图示。Figure 21 is an XRPD diagram of nitrate.
图22为大鼠PK实验图示。Figure 22 is a schematic diagram of a rat PK experiment.
具体实施方式detailed description
实验仪器laboratory apparatus
物理化学检测仪器的一些参数Some parameters of physical and chemical testing instruments
Figure PCTCN2020082042-appb-000040
Figure PCTCN2020082042-appb-000040
Figure PCTCN2020082042-appb-000041
Figure PCTCN2020082042-appb-000041
液相分析条件Liquid phase analysis conditions
仪器与设备Instruments and equipment
仪器名称equipment name 型号model
分析天平Analytical Balances Sartorius BSA224S-CWSartorius BSA224S-CW
纯水机Pure water machine Milli-Q Plus，MilliporeMilli-Q Plus, Millipore
高效液相色谱仪High performance liquid chromatography Agilent1260Agilent1260
泵Pump Agilent G1311BAgilent G1311B
进样器Injector G1329BG1329B
柱温箱Column thermostat G1316AG1316A
检测器Detector G1315DG1315D
色谱条件Chromatographic conditions
色谱柱：XBridge TM(C18，3.5μm，4.6*150mm) Column: XBridge TM (C18, 3.5μm, 4.6*150mm)
流速：1mL/minFlow rate: 1mL/min
柱温：50℃Column temperature: 50℃
检测波长：255nmDetection wavelength: 255nm
进样体积：5.0μLInjection volume: 5.0μL
运行时间：35minRunning time: 35min
稀释剂：NMP-水(v/v，1:4)Thinner: NMP-water (v/v, 1:4)
流动相：A：水(0.05％三氟乙酸)；B：乙腈(0.05％三氟乙酸)Mobile phase: A: water (0.05% trifluoroacetic acid); B: acetonitrile (0.05% trifluoroacetic acid)
T(min)T(min) B(％)B(%)
0.000.00 55
14.0014.00 3333
25.0025.00 3333
30.0030.00 9595
30.2030.20 55
35.0035.00 55
自由碱的制备Free base preparation
实施例1Example 1
(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢吡咯并[3,2-f]吲唑-6-甲酰胺的制备(R)-N-(1-Phenylethyl)-3-(pyridin-4-yl)-1,7-dihydropyrrolo[3,2-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000042
Figure PCTCN2020082042-appb-000042
第一步：3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)丙烯酰酸甲酯的制备The first step: preparation of methyl 3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)acryloyl acid
Figure PCTCN2020082042-appb-000043
Figure PCTCN2020082042-appb-000043
6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-甲醛(350mg，0.68mmol)和甲氧甲酰基亚甲基三苯基膦(344mg，1.03mmol)溶于甲苯(20mL)中，加热至105℃搅拌2小时。反应完，浓缩反应液后柱层析[洗脱剂:石油醚～石油醚/乙酸乙酯(60:40)]纯化得到淡青色固体产物3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)丙烯酰酸甲酯(200mg，产率51％)。6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazole-5-carbaldehyde (350mg, 0.68mmol) and methoxyformylmethylenetriphenylphosphine ( 344mg, 1.03mmol) was dissolved in toluene (20mL), heated to 105°C and stirred for 2 hours. After the reaction is complete, the reaction solution is concentrated and purified by column chromatography [eluent: petroleum ether ~ petroleum ether/ethyl acetate (60:40)] to obtain a pale blue solid product 3-(6-nitro-3-(pyridine-4) -Yl)-1-tritylmethyl-1H-indazol-5-yl)acrylic acid methyl ester (200 mg, yield 51%).
MS m/z(ESI):567.1[M+H] +. MS m/z(ESI): 567.1[M+H] + .
第二步：3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)丙烯酸制备Step 2: Preparation of 3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)acrylic acid
Figure PCTCN2020082042-appb-000044
Figure PCTCN2020082042-appb-000044
3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)丙烯酰酸甲酯(150mg，0.26mmol)和一水合氢氧化锂(22mg，0.53mmol)溶于四氢呋喃(8mL)和水(2mL)中，室温搅拌4小时。反应完加0.5N醋酸水溶液(30mL)淬灭，乙酸乙酯(30mL*2)萃取，有机相用氯化钠水溶液(20mL)洗涤，硫酸钠干燥，浓缩得到橘黄色固体产物3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)丙烯酸(150mg，产率100％)。3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)acrylic acid methyl ester (150mg, 0.26mmol) and hydrogen monohydrate Lithium oxide (22 mg, 0.53 mmol) was dissolved in tetrahydrofuran (8 mL) and water (2 mL), and stirred at room temperature for 4 hours. After the reaction, it was quenched with 0.5N acetic acid aqueous solution (30mL), extracted with ethyl acetate (30mL*2), the organic phase was washed with sodium chloride aqueous solution (20mL), dried over sodium sulfate and concentrated to obtain an orange solid product 3-(6- Nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)acrylic acid (150 mg, yield 100%).
MS m/z(ESI):553.1[M+H] +。 MS m/z (ESI): 553.1 [M+H] + .
第三步：(R)-3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)-N-(1-苯基乙基) 丙烯酰基酰胺的制备The third step: (R)-3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)-N-(1-benzene (Ethyl) acrylamide preparation
Figure PCTCN2020082042-appb-000045
Figure PCTCN2020082042-appb-000045
往3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)丙烯酸(150mg，0.27mmol)，(R)-1-苯基乙烷-1-胺(49mg，0.40mmol)和O-(7-氮杂苯并三唑-1-基)-N,N,N',N'-四甲基脲六氟磷酸盐(207mg，0.54mmol)的四氢呋喃(20mL)溶液里加三乙胺(816mg，0.81mmol)，加完室温搅拌16小时。往反应液里加乙酸乙酯(60mL)，按顺序用0.5N醋酸(30mL*2)，碳酸氢钠水溶液(20mL)，氯化钠水溶液(20mL)洗涤，硫酸钠干燥，浓缩得到黄色固体产物(R)-3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)-N-(1-苯基乙基)丙烯酰基酰胺(150mg，产率84％)。To 3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)acrylic acid (150mg, 0.27mmol), (R)-1- Phenylethane-1-amine (49mg, 0.40mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate Add triethylamine (816 mg, 0.81 mmol) to a solution of tetrahydrofuran (20 mL) (207 mg, 0.54 mmol), and stir at room temperature for 16 hours. Ethyl acetate (60mL) was added to the reaction solution, washed with 0.5N acetic acid (30mL*2), sodium bicarbonate aqueous solution (20mL), sodium chloride aqueous solution (20mL), dried over sodium sulfate, and concentrated to obtain a yellow solid product R)-3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)-N-(1-phenylethyl)propene Acylamide (150 mg, 84% yield).
MS m/z(ESI):656.2[M+H] +. MS m/z(ESI): 656.2[M+H] + .
第四步：(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢吡咯并[3,2-f]吲唑-6-甲酰胺的制备The fourth step: (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydropyrrolo[3,2- f] Preparation of indazole-6-carboxamide
Figure PCTCN2020082042-appb-000046
Figure PCTCN2020082042-appb-000046
(R)-3-(6-硝基-3-(吡啶-4-基)-1-三苯代甲基-1H-吲唑-5-基)-N-(1-苯基乙基)丙烯酰基酰胺(130mg，0.19mmol)在亚磷酸三乙酯(3mL)中140℃微波45分钟。浓缩反应液后柱层析纯化[洗脱剂：石油醚～石油醚/乙酸乙酯(40:60)]得到黄色油状物产物(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢吡咯并[3,2-f]吲唑-6-甲酰胺(50mg，产率40％)。(R)-3-(6-nitro-3-(pyridin-4-yl)-1-tritylmethyl-1H-indazol-5-yl)-N-(1-phenylethyl) Acrylamide (130 mg, 0.19 mmol) was microwaved in triethyl phosphite (3 mL) at 140°C for 45 minutes. The reaction solution was concentrated and purified by column chromatography [eluent: petroleum ether ~ petroleum ether/ethyl acetate (40:60)] to obtain the product (R)-N-(1-phenylethyl)-3- (Pyridin-4-yl)-1-tritylmethyl-1,7-dihydropyrrolo[3,2-f]indazole-6-carboxamide (50 mg, yield 40%).
MS m/z(ESI):624.2[M+H] +. MS m/z(ESI): 624.2[M+H] + .
第五步：(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢吡咯并[3,2-f]吲唑-6-甲酰胺的制备The fifth step: (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydropyrrolo[3,2-f]indazole-6-methan Preparation of amide
Figure PCTCN2020082042-appb-000047
Figure PCTCN2020082042-appb-000047
往(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢吡咯并[3,2-f]吲唑-6-甲酰胺(50mg，0.08mmol)和三乙基硅烷(19mg，0.16mmol)的二氯甲烷(2mL)溶液里加三氟乙酸(4mL)，室温搅拌2小时。旋干反应液，加甲醇 溶解，用氨水调至碱性，再薄层层析(展开剂：CH 2Cl 2/MeOH＝10/1)纯化得到黄色固体产物(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢吡咯并[3,2-f]吲唑-6-甲酰胺(6mg，产率20％)。 To (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydropyrrolo[3,2-f]indole Trifluoroacetic acid (4 mL) was added to a dichloromethane (2 mL) solution of azole-6-carboxamide (50 mg, 0.08 mmol) and triethylsilane (19 mg, 0.16 mmol), and stirred at room temperature for 2 hours. The reaction solution was spin-dried, dissolved in methanol, adjusted to alkaline with ammonia, and purified by thin layer chromatography (developing solvent: CH 2 Cl 2 /MeOH = 10/1) to obtain a yellow solid product (R)-N-(1- Phenylethyl)-3-(pyridin-4-yl)-1,7-dihydropyrrolo[3,2-f]indazole-6-carboxamide (6 mg, yield 20%).
1H NMR(400MHz,MeOD)δ8.66(d,J＝4Hz,2H),8.41(s,1H),8.12(d,J＝4Hz,2H),7.53(s,1H),7.44(d,J＝8Hz,2H),7.39–7.30(m,3H),7.26–7.22(m,1H),5.29(q,J＝8Hz,1H),1.61(d,J＝8Hz,3H). 1 H NMR (400MHz, MeOD) δ8.66 (d, J = 4Hz, 2H), 8.41 (s, 1H), 8.12 (d, J = 4Hz, 2H), 7.53 (s, 1H), 7.44 (d, J=8Hz, 2H), 7.39-7.30(m, 3H), 7.26-7.22(m, 1H), 5.29(q, J=8Hz, 1H), 1.61(d, J=8Hz, 3H).
MS m/z(ESI):382.1[M+H] +. MS m/z(ESI): 382.1[M+H] + .
实施例2Example 2
(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-Phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000048
Figure PCTCN2020082042-appb-000048
第一步：3-溴-5,6-二硝基-1-三苯甲基-1H-吲唑的制备Step 1: Preparation of 3-bromo-5,6-dinitro-1-trityl-1H-indazole
Figure PCTCN2020082042-appb-000049
Figure PCTCN2020082042-appb-000049
3-溴-5,6-二硝基-1H-吲唑(5.0g，17.42mmol)、碳酸铯(8.51g，26.12mmol)、DMF(50mL)搅拌，分批加入三苯基氯甲烷(5.83g，20.91mmol)，室温搅拌过夜。加水250mL，搅拌1h，过滤，水洗，得固体产物3-溴-5,6-二硝基-1-三苯甲基-1H-吲唑(11.47g，湿重)。不用干燥，直接用于下一步反应。3-Bromo-5,6-dinitro-1H-indazole (5.0g, 17.42mmol), cesium carbonate (8.51g, 26.12mmol), DMF (50mL) were stirred, and triphenylchloromethane (5.83 g, 20.91mmol), stirred at room temperature overnight. Add 250 mL of water, stir for 1 h, filter and wash with water to obtain the solid product 3-bromo-5,6-dinitro-1-trityl-1H-indazole (11.47g, wet weight). It is directly used in the next reaction without drying.
第二步：5,6-二硝基-3-(吡啶-4-基)-1-三苯甲基-1H-吲唑的制备Step 2: Preparation of 5,6-Dinitro-3-(pyridin-4-yl)-1-trityl-1H-indazole
Figure PCTCN2020082042-appb-000050
Figure PCTCN2020082042-appb-000050
3-溴-5,6-二硝基-1-三苯甲基-1H-吲唑(11.47g，21.66mmol)、4-吡啶硼酸(5.33g，43.36mmol)、Pd(dppf)Cl 2(1.59g，4.33mmol)、碳酸钾(6.0g，14mmol)、二氧六环/水(125mL/25mL)于氮气保护下80℃下搅拌5小时，冷却至室温，加水，乙酸乙酯提取，柱层析纯化得固体产物5,6-二硝基-3-(吡啶-4-基)-1-三苯甲基-1H-吲唑(7.50g，两步产率81％)。 3-Bromo-5,6-dinitro-1-trityl-1H-indazole (11.47g, 21.66mmol), 4-pyridineboronic acid (5.33g, 43.36mmol), Pd(dppf)Cl 2 ( 1.59g, 4.33mmol), potassium carbonate (6.0g, 14mmol), dioxane/water (125mL/25mL) were stirred at 80°C for 5 hours under nitrogen protection, cooled to room temperature, water was added, and extracted with ethyl acetate. Chromatographic purification gave the solid product 5,6-dinitro-3-(pyridin-4-yl)-1-trityl-1H-indazole (7.50 g, 81% yield in two steps).
MS m/z(ESI):528.1[M+H] +. MS m/z(ESI):528.1[M+H] + .
第三步：3-(吡啶-4-基)-1-三苯甲基-1H-吲唑-5,6-二胺的制备The third step: Preparation of 3-(pyridin-4-yl)-1-trityl-1H-indazole-5,6-diamine
Figure PCTCN2020082042-appb-000051
Figure PCTCN2020082042-appb-000051
5,6-二硝基-3-(吡啶-4-基)-1-三苯甲基-1H-吲唑(2.5g，4.74mmol)、10％Pd/C(200mg)、THF(150mL)室温氢化过夜，过滤，浓缩至干得棕色固体产物3-(吡啶-4-基)-1-三苯甲基-1H-吲唑-5,6-二胺，直接用于下一步反应。5,6-Dinitro-3-(pyridin-4-yl)-1-trityl-1H-indazole (2.5g, 4.74mmol), 10% Pd/C (200mg), THF (150mL) Hydrogenate at room temperature overnight, filter, and concentrate to dry to obtain the brown solid product 3-(pyridin-4-yl)-1-trityl-1H-indazole-5,6-diamine, which is directly used in the next reaction.
MS m/z(ESI):468.2[M+H] +. MS m/z(ESI): 468.2[M+H] + .
第四步：3-(吡啶-4-基)-6-(三氯甲基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑的制备Step 4: Preparation of 3-(pyridin-4-yl)-6-(trichloromethyl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole
Figure PCTCN2020082042-appb-000052
Figure PCTCN2020082042-appb-000052
3-(吡啶-4-基)-1-三苯甲基-1H-吲唑-5,6-二胺(上一步所得)、2,2,2-三氯乙酰亚胺甲酯(920mg，5.22mmol)、醋酸(25mL)室温搅拌过夜，加水100mL，搅拌1小时，过滤，水洗得棕色固体产物3-(吡啶-4-基)-6-(三氯甲基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑(4.32g，湿重)。直接用于下一步反应。3-(pyridin-4-yl)-1-trityl-1H-indazole-5,6-diamine (obtained in the previous step), methyl 2,2,2-trichloroacetimide (920mg, 5.22mmol), acetic acid (25mL), stir overnight at room temperature, add 100mL of water, stir for 1 hour, filter, and wash with water to obtain the brown solid product 3-(pyridin-4-yl)-6-(trichloromethyl)-1-trityl Base-1,7-dihydroimidazo[4,5-f]indazole (4.32g, wet weight). Used directly in the next reaction.
MS m/z(ESI):594.1，596.1[M+H] +. MS m/z(ESI): 594.1, 596.1[M+H] + .
第五步：3-(吡啶-4-基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯的制备Step 5: Preparation of 3-(pyridin-4-yl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester
Figure PCTCN2020082042-appb-000053
Figure PCTCN2020082042-appb-000053
3-(吡啶-4-基)-6-(三氯甲基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑(4.32g)、甲醇(50mL)回流过夜，浓缩至干。加异丙醚搅拌30min，过滤，异丙醚洗得棕色固体粗品3-(吡啶-4-基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯(2.44g)。3-(pyridin-4-yl)-6-(trichloromethyl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole (4.32g), methanol ( 50 mL) reflux overnight and concentrate to dryness. Add isopropyl ether and stir for 30min, filter, and wash with isopropyl ether to obtain crude brown solid 3-(pyridin-4-yl)-1-trityl-1,7-dihydroimidazo[4,5-f]indole Methyl oxazole-6-carboxylate (2.44 g).
MS m/z(ESI):536.2[M+H] +. MS m/z(ESI): 536.2[M+H] + .
第六步：(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺The sixth step: (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5- f]Indazole-6-carboxamide
Figure PCTCN2020082042-appb-000054
Figure PCTCN2020082042-appb-000054
将3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯(100mg，0.19mmol)置于微波反应管中，加入(R)-1-苯基乙烷-1-胺(1.0mL)，加热至150℃反应60min。待冷却至室温后，向反应液中加入20mL乙酸乙酯，乙酸乙酯层用饱和氯化铵溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥后柱层析，得粗产物(80mg)，直接用于下一步反应。Place 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester (100mg, 0.19mmol) In the microwave reaction tube, add (R)-1-phenylethane-1-amine (1.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution. The ethyl acetate layer was washed with saturated ammonium chloride solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate and column chromatography was used to obtain the crude product ( 80mg), directly used in the next reaction.
MS m/z(ESI):625.1[M+H] +. MS m/z(ESI): 625.1[M+H] + .
第七步：(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The seventh step: (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-methan Preparation of amide
Figure PCTCN2020082042-appb-000055
Figure PCTCN2020082042-appb-000055
将上步所得粗品(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(80mg)溶于4mL二氯甲烷，同时加入4mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥。所得有机溶剂旋干后通过制备色谱纯化，得黄色固体产物(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(15mg，两步产率21％)。The crude product (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide (80 mg) was dissolved in 4 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added at the same time. After 3 hours of reaction at room temperature, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by preparative chromatography to obtain the yellow solid product (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4 ,5-f]Indazole-6-carboxamide (15 mg, 21% yield for two steps).
1H NMR(400MHz,DMSO-d 6)δ:13.45-13.39(m,1H),13.21-13.08(m,1H),9.43–9.33(m,1H),8.72(s,2H),8.44–8.04(m,3H),7.99-7.87(m,1H),7.58(m,2H),7.36–7.32(m,2H),7.26-7.22(m,1H),5.25–5.21(m,1H),1.57(d,J＝7.0Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ: 13.45-13.39 (m, 1H), 13.21-13.08 (m, 1H), 9.43-9.33 (m, 1H), 8.72 (s, 2H), 8.44-8.04 (m,3H),7.99-7.87(m,1H),7.58(m,2H),7.36--7.32(m,2H),7.26-7.22(m,1H),5.25-5.21(m,1H),1.57 (d,J=7.0Hz,3H).
MS m/z(ESI):383.1[M+H] +. MS m/z(ESI): 383.1[M+H] + .
实施例3Example 3
N-(3,4-二甲氧苄基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Preparation of N-(3,4-Dimethoxybenzyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000056
Figure PCTCN2020082042-appb-000056
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(3,4-二甲氧基苯基)甲胺为原料参考实施例2步骤六和步骤七得到N-(3,4-二甲氧苄基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (3,4-di (Methoxyphenyl) methylamine is used as the raw material. Refer to Example 2 Step 6 and Step 7 to obtain N-(3,4-Dimethoxybenzyl)-3-(pyridin-4-yl)-1,7-dihydro Imidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.48(s,1H),9.37(d,J＝8.6Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.72(t,J＝10.2Hz,1H),6.93–6.84(m,2H),6.89–6.78(m,1H),4.72(dt,J＝10.0,1.0Hz,2H),3.82(d,J＝8.5Hz,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.48 (s, 1H), 9.37 (d, J = 8.6 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s, 1H), 7.96–7.89 (m, 2H), 7.72 (t, J = 10.2 Hz, 1H), 6.93–6.84 (m, 2H), 6.89–6.78 (m, 1H), 4.72 (dt, J = 10.0,1.0Hz,2H),3.82(d,J=8.5Hz,6H).
MS m/z(ESI):429.1[M+H] +. MS m/z(ESI): 429.1[M+H] + .
实施例4Example 4
N-(3-氯-4-氟苯甲基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Preparation of N-(3-chloro-4-fluorobenzyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000057
Figure PCTCN2020082042-appb-000057
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(3-氯-4-氟苯基)甲胺为原料参考实施例2步骤六和步骤七得到N-(3-氯-4-氟苯甲基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (3-chloro-4 -Fluorophenyl)methylamine is used as a raw material. Refer to Example 2 Steps 6 and 7 to obtain N-(3-chloro-4-fluorobenzyl)-3-(pyridin-4-yl)-1,7-dihydro Imidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),9.38(d,J＝8.6Hz,1H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.72(t,J＝10.2Hz,1H),7.38(dd,J＝7.4,4.9Hz,1H),7.14(ddq,J＝9.1,7.9,1.2Hz,2H),4.73(dt,J＝10.1,1.0Hz,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47 (s, 1H), 9.38 (d, J = 8.6 Hz, 1H), 8.13 (s, 1H), 7.99 (s, 1H), 7.96–7.89 ( m, 2H), 7.72 (t, J = 10.2 Hz, 1H), 7.38 (dd, J = 7.4, 4.9 Hz, 1H), 7.14 (ddq, J = 9.1, 7.9, 1.2 Hz, 2H), 4.73 (dt , J = 10.1, 1.0 Hz, 2H).
MS m/z(ESI):421.2[M+H] +. MS m/z(ESI): 421.2[M+H] + .
实施例5Example 5
(R)-3-(吡啶-4-基)-N-(1-(p-苯甲基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-3-(pyridin-4-yl)-N-(1-(p-benzyl)ethyl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000058
Figure PCTCN2020082042-appb-000058
第一步：(R)-3-(吡啶-4-基)-N-(1-(p-苯甲基)乙基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-3-(pyridin-4-yl)-N-(1-(p-phenylmethyl)ethyl)-1-tritylmethyl-1,7-dihydroimidazo Preparation of [4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000059
Figure PCTCN2020082042-appb-000059
将甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(100mg,0.19mmol)置于微波反应管中，加入(R)-1-(p-苯甲基)乙烷-1-胺(1.0mL)，加热至150℃反应60min。待冷却至室温后，向反应液中加入20mL乙酸乙酯，乙酸乙酯层用饱和氯化铵溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥后柱层析，得粗产物(80mg,产率67％)，直接用于下一步反应。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (100mg, 0.19mmol) Place in a microwave reaction tube, add (R)-1-(p-benzyl)ethane-1-amine (1.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution. The ethyl acetate layer was washed with saturated ammonium chloride solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate and column chromatography was used to obtain the crude product ( 80mg, yield 67%), directly used in the next reaction.
MS m/z(ESI):639.3[M+H] +. MS m/z(ESI): 639.3[M+H] + .
第二步：(R)-3-(吡啶-4-基)-N-(1-(p-苯甲基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-3-(pyridin-4-yl)-N-(1-(p-benzyl)ethyl)-1,7-dihydroimidazo[4,5-f]indole Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000060
Figure PCTCN2020082042-appb-000060
将(R)-3-(吡啶-4-基)-N-(1-(p-苯甲基)乙基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺粗品(80mg)溶于4mL二氯甲烷，同时加入4mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥。所得有机溶剂旋干后通过制备色谱纯化，得黄色固体产物(R)-3-(吡啶-4-基)-N-(1-(p-苯甲基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(10mg,产率20％)。The (R)-3-(pyridin-4-yl)-N-(1-(p-benzyl)ethyl)-1-tritylmethyl-1,7-dihydroimidazo[4, 5-f] Indazole-6-carboxamide crude product (80 mg) was dissolved in 4 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added at the same time. After reacting for 3 hours at room temperature, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by preparative chromatography to obtain the yellow solid product (R)-3-(pyridin-4-yl)-N-(1-(p-benzyl)ethyl)-1,7-di Hydroimidazo[4,5-f]indazole-6-carboxamide (10 mg, yield 20%).
1H NMR(400MHz,MeOD)δ8.57(d,J＝4Hz,2H),8.37(s,0.6H),8.15(s,0.4H),8.02(d,J＝4Hz,2H),7.79(s,0.4H),7.56(s,0.6H),7.25(d,J＝8Hz,2H),7.08(d,J＝8Hz,2H),5.12-5.16(m,1H),2.22(s,3H),1.52(d,J＝8Hz,3H). 1 H NMR (400MHz, MeOD) δ8.57 (d, J = 4Hz, 2H), 8.37 (s, 0.6H), 8.15 (s, 0.4H), 8.02 (d, J = 4Hz, 2H), 7.79 ( s, 0.4H), 7.56 (s, 0.6H), 7.25 (d, J = 8 Hz, 2H), 7.08 (d, J = 8 Hz, 2H), 5.12-5.16 (m, 1H), 2.22 (s, 3H ), 1.52(d,J=8Hz,3H).
MS m/z(ESI):397.1[M+H] +. MS m/z(ESI): 397.1[M+H] + .
实施例6Example 6
(R)-N-(1-(4-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(4-Fluorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000061
Figure PCTCN2020082042-appb-000061
第一步：(R)-N-(1-(4-氟苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-N-(1-(4-fluorophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo Preparation of [4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000062
Figure PCTCN2020082042-appb-000062
甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(1.5g，2.80mmol)、(R)-1-(4-氟苯基)乙烷-1-胺(2.5mL)于150℃微波搅拌1h，冷却，浓缩至干，柱层析纯化得粘稠物粗品，直接用于下一步反应。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (1.5g, 2.80mmol) , (R)-1-(4-Fluorophenyl)ethane-1-amine (2.5mL) was stirred at 150℃ in microwave for 1h, cooled, concentrated to dryness, purified by column chromatography to obtain the crude viscous product, which was used directly Next reaction.
MS m/z(ESI):643.2[M+H] +. MS m/z(ESI):643.2[M+H] + .
第二步：(R)-N-(1-(4-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-N-(1-(4-fluorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indole Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000063
Figure PCTCN2020082042-appb-000063
将上步产物溶于二氯甲烷(5mL)，加三氟乙酸(5mL)，室温搅拌2h，浓缩至干，柱层析纯化得黄色固体(R)-N-(1-(4-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(603mg，二步产率54％)。 1H NMR(400MHz,MeOD)δ8.89(s,1H),8.55(s,1H),8.18(s,1H),7.85(s,1H),7.28(dd,J＝15.0,8.3Hz,1H),6.80(ddd,J＝20.8,17.5,10.7Hz,3H),6.28(dd,J＝16.7,1.8Hz,1H),5.80(dd,J＝10.6,1.7Hz,1H),4.56(m,1H),3.98–3.81(m,3H). The product from the previous step was dissolved in dichloromethane (5mL), added with trifluoroacetic acid (5mL), stirred at room temperature for 2h, concentrated to dryness, and purified by column chromatography to obtain a yellow solid (R)-N-(1-(4-fluorobenzene) (Yl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (603 mg, 54% yield in two steps). 1 H NMR (400MHz, MeOD) δ 8.89 (s, 1H), 8.55 (s, 1H), 8.18 (s, 1H), 7.85 (s, 1H), 7.28 (dd, J = 15.0, 8.3 Hz, 1H ), 6.80 (ddd, J = 20.8, 17.5, 10.7 Hz, 3H), 6.28 (dd, J = 16.7, 1.8 Hz, 1H), 5.80 (dd, J = 10.6, 1.7 Hz, 1H), 4.56 (m, 1H), 3.98--3.81 (m, 3H).
MS m/z(ESI):401.1[M+H] +. MS m/z(ESI): 401.1[M+H] + .
实施例7Example 7
N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Preparation of N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000064
Figure PCTCN2020082042-appb-000064
第一步：N-(1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: N-(1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole Preparation of -6-formamide
Figure PCTCN2020082042-appb-000065
Figure PCTCN2020082042-appb-000065
甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(100mg，0.18mmol)、1-苯基乙烷-1-胺(1.5mL)于150℃微波搅拌1h，冷却，浓缩至干，柱层析纯化得N-(1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺粗品(27mg)。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (100mg, 0.18mmol), 1-Phenylethane-1-amine (1.5mL) was stirred at 150℃ in microwave for 1h, cooled, concentrated to dryness, and purified by column chromatography to obtain N-(1-phenylethyl)-3-(pyridine-4- Yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide crude product (27mg).
MS m/z(ESI):625.2[M+H] +. MS m/z(ESI): 625.2[M+H] + .
第二步：N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Step 2: Preparation of N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000066
Figure PCTCN2020082042-appb-000066
将N-(1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺粗品(27mg)溶于二氯甲烷(2mL)，加三氟乙酸(2mL)，室温搅拌2h，浓缩至干，柱层析纯化得黄色固体N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(5mg，二步产率7％)。The N-(1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6- The crude formamide (27mg) was dissolved in dichloromethane (2mL), added with trifluoroacetic acid (2mL), stirred at room temperature for 2h, concentrated to dryness, purified by column chromatography to obtain a yellow solid N-(1-phenylethyl)-3 -(Pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (5 mg, yield 7% in two steps).
1H NMR(400MHz,DMSO-d 6)δ13.48(s,1H),9.37(d,J＝8.6Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.36–7.27(m,2H),7.31–7.20(m,3H),6.75(d,J＝9.3Hz,1H),5.20(dqt,J＝8.7,6.8,0.9Hz,1H),1.57(d,J＝6.7Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.48 (s, 1H), 9.37 (d, J = 8.6 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s,1H),7.96–7.89(m,2H),7.36–7.27(m,2H),7.31–7.20(m,3H), 6.75(d,J=9.3Hz,1H), 5.20(dqt,J= 8.7, 6.8, 0.9Hz, 1H), 1.57 (d, J=6.7Hz, 3H).
MS m/z(ESI):383.1[M+H] +. MS m/z(ESI): 383.1[M+H] + .
实施例8Example 8
(S)-N-(2-甲氧基-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(2-Methoxy-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6 -Formamide preparation
Figure PCTCN2020082042-appb-000067
Figure PCTCN2020082042-appb-000067
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(S)-2-甲氧基-1-苯基乙-1-胺为原料参考实施例2步骤六和步骤七得到(S)-N-(2-甲氧基-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (S)-2- (S)-N-(2-methoxy-1-phenylethyl)-3-(pyridine) was obtained by using methoxy-1-phenylethyl-1-amine as the raw material. Refer to Example 2 Steps 6 and 7 -4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.48(s,1H),9.37(d,J＝8.6Hz,1H),8.72(d,J＝5.2Hz,2H),8.34(s,2H),8.04(d,J＝5.1Hz,2H),7.82–7.60(m,1H),7.50(d,J＝7.4Hz,2H),7.36(t,J＝7.5Hz,2H),7.28(t,J＝7.3Hz,1H),5.31(td,J＝8.4,4.8Hz,1H),3.87(t,J＝9.2Hz,2H),3.64(d,J＝5.0Hz,3H). 1 H NMR(400MHz,DMSO-d 6 )δ13.48(s,1H), 9.37(d,J=8.6Hz,1H), 8.72(d,J=5.2Hz,2H), 8.34(s,2H) ,8.04(d,J=5.1Hz,2H),7.82-7.60(m,1H),7.50(d,J=7.4Hz,2H),7.36(t,J=7.5Hz,2H),7.28(t, J = 7.3Hz, 1H), 5.31 (td, J = 8.4, 4.8 Hz, 1H), 3.87 (t, J = 9.2 Hz, 2H), 3.64 (d, J = 5.0 Hz, 3H).
MS m/z(ESI):413.1[M+H] +. MS m/z(ESI): 413.1[M+H] + .
实施例9Example 9
(R)-N-(1-(4-硝基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(4-nitrophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6 -Formamide preparation
Figure PCTCN2020082042-appb-000068
Figure PCTCN2020082042-appb-000068
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-(4-硝基苯基)乙-1-胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(4-硝基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- (4-Nitrophenyl) ethyl-1-amine is used as raw material. Refer to Example 2 Steps 6 and 7 to obtain (R)-N-(1-(4-nitrophenyl)ethyl)-3-(pyridine -4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.44-13.39(m,1H),13.20-13.08(m,1H),9.43–9.33(m,1H),8.68–8.62(m,2H),8.22–8.14(m,2H),8.00–7.94(m,2H),7.70(s,2H),7.62–7.54(m,2H),5.27–5.22(m,1H),1.56(d,J＝6.4Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.44-13.39 (m, 1H), 13.20-13.08 (m, 1H), 9.43-9.33 (m, 1H), 8.68-8.62 (m, 2H), 8.22 –8.14(m,2H),8.00–7.94(m,2H),7.70(s,2H),7.62–7.54(m,2H),5.27–5.22(m,1H),1.56(d,J=6.4Hz ,3H).
MS m/z(ESI):428.1[M+H] +. MS m/z(ESI): 428.1[M+H] + .
实施例10Example 10
(R)-N-(1-(4-溴苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(4-Bromophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000069
Figure PCTCN2020082042-appb-000069
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-(4-溴苯基)乙-1-胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(4-溴苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- (4-Bromophenyl)ethyl-1-amine is used as the raw material. Refer to Example 2 in step six and step seven to obtain (R)-N-(1-(4-bromophenyl)ethyl)-3-(pyridine-4 -Yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),9.48(d,J＝7.9Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.50–7.42(m,2H),7.26–7.19(m,2H),6.75(d,J＝9.3Hz,1H),5.22(dqt,J＝8.8,6.8,1.0Hz,1H),1.57(d,J＝6.7Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47 (s, 1H), 9.48 (d, J = 7.9 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s,1H),7.96–7.89(m,2H),7.50–7.42(m,2H),7.26–7.19(m,2H), 6.75(d,J=9.3Hz,1H),5.22(dqt,J= 8.8, 6.8, 1.0Hz, 1H), 1.57 (d, J=6.7Hz, 3H).
MS m/z(ESI):461.1，463.0[M+H] +. MS m/z(ESI): 461.1, 463.0[M+H] + .
实施例11Example 11
(R)-3-(吡啶-4-基)-N-(1-(3-(三氟甲基)苯基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-3-(pyridin-4-yl)-N-(1-(3-(trifluoromethyl)phenyl)ethyl)-1,7-dihydroimidazo[4,5-f] Preparation of indazole-6-carboxamide
Figure PCTCN2020082042-appb-000070
Figure PCTCN2020082042-appb-000070
第一步：(R)-3-(吡啶-4-基)-N-(1-(4-(三氟甲基)苯基)乙基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-3-(pyridin-4-yl)-N-(1-(4-(trifluoromethyl)phenyl)ethyl)-1-tritylmethyl-1,7 -Preparation of dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000071
Figure PCTCN2020082042-appb-000071
将甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(100mg,0.19mmol)置于微波反应管中，加入(R)-1-(4-(三氟甲基)苯基)乙烷-1-胺(1.0mL)，加热至150℃反应60min。待冷却至室温后，向反应液中加入20mL乙酸乙酯，乙酸乙酯层用饱和氯化铵溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥后柱层析，得粗产物(80mg，产率67％)，直接用于下一步反应。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (100mg, 0.19mmol) Place in a microwave reaction tube, add (R)-1-(4-(trifluoromethyl)phenyl)ethane-1-amine (1.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution. The ethyl acetate layer was washed with saturated ammonium chloride solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate and column chromatography was used to obtain the crude product ( 80mg, yield 67%), directly used in the next reaction.
MS m/z(ESI):693.3[M+H] +. MS m/z(ESI): 693.3[M+H] + .
第二步：(R)-3-(吡啶-4-基)-N-(1-(3-(三氟甲基)苯基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-3-(pyridin-4-yl)-N-(1-(3-(trifluoromethyl)phenyl)ethyl)-1,7-dihydroimidazo[4, 5-f]Indazole-6-carboxamide preparation
Figure PCTCN2020082042-appb-000072
Figure PCTCN2020082042-appb-000072
将(R)-3-(吡啶-4-基)-N-(1-(4-(三氟甲基)苯基)乙基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺粗品(80mg)溶于4mL二氯甲烷，同时加入4mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干 燥。所得有机溶剂旋干后通过制备色谱纯化，得黄色固体产物(R)-3-(吡啶-4-基)-N-(1-(3-(三氟甲基)苯基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(10mg，产率19％)。Add (R)-3-(pyridin-4-yl)-N-(1-(4-(trifluoromethyl)phenyl)ethyl)-1-trityl-1,7-dihydro The crude imidazo[4,5-f] indazole-6-carboxamide (80 mg) was dissolved in 4 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added at the same time. After reacting for 3 hours at room temperature, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried over anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by preparative chromatography to obtain the yellow solid product (R)-3-(pyridin-4-yl)-N-(1-(3-(trifluoromethyl)phenyl)ethyl)- 1,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide (10 mg, yield 19%).
1H NMR(400MHz,MeOD)δ8.59(d,J＝4Hz,2H),8.39(s,0.6H),8.15(s,0.4H),8.06(d,J＝4Hz,2H),7.79(s,0.4H),7.56(s,0.6H),7.49(d,J＝8Hz,2H),7.21(d,J＝8Hz,2H),5.12-5.16(m,1H),1.52(d,J＝8Hz,3H). 1 H NMR (400MHz, MeOD) δ8.59 (d, J = 4Hz, 2H), 8.39 (s, 0.6H), 8.15 (s, 0.4H), 8.06 (d, J = 4Hz, 2H), 7.79 ( s, 0.4H), 7.56 (s, 0.6H), 7.49 (d, J = 8 Hz, 2H), 7.21 (d, J = 8 Hz, 2H), 5.12-5.16 (m, 1H), 1.52 (d, J ＝8Hz, 3H).
MS m/z(ESI):451.1[M+H] +. MS m/z(ESI): 451.1[M+H] + .
实施例12Example 12
(R)-N-(1-苯基丙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-Phenylpropyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000073
Figure PCTCN2020082042-appb-000073
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-苯基丙-1-胺胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-苯基丙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- Phenylpropan-1-amine amine was used as the raw material. Refer to Example 2 in step six and step seven to obtain (R)-N-(1-phenylpropyl)-3-(pyridin-4-yl)-1,7-di Hydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.56-13.03(m,2H),9.25-9.14(m,1H),8.72(d,J＝4Hz,2H),8.50(s,0.6H),8.06-7.90(m,2.7H),7.57(s,0.6H),7.45(d,J＝8Hz,2H),7.35(t,J＝8Hz,2H),7.28-7.23(m,1H),5.14-5.09(m,1H),1.75-1.70(m,2H),0.95-0.88(m,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.56-13.03 (m, 2H), 9.25-9.14 (m, 1H), 8.72 (d, J = 4Hz, 2H), 8.50 (s, 0.6H), 8.06-7.90(m,2.7H),7.57(s,0.6H),7.45(d,J=8Hz,2H),7.35(t,J=8Hz,2H),7.28-7.23(m,1H),5.14 -5.09 (m, 1H), 1.75-1.70 (m, 2H), 0.95-0.88 (m, 3H).
MS m/z(ESI):397.1[M+H] +. MS m/z(ESI): 397.1[M+H] + .
实施例13Example 13
(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(3-Bromophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000074
Figure PCTCN2020082042-appb-000074
第一步：(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-N-(1-(3-bromophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo Preparation of [4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000075
Figure PCTCN2020082042-appb-000075
甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(100mg，0.18mmol)、(R)-1-(3-溴苯基)乙烷-1-胺(1mL)于150℃微波搅拌1h，冷却，浓缩至干，柱层析纯化粗品(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(120mg)。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (100mg, 0.18mmol), (R)-1-(3-Bromophenyl)ethane-1-amine (1mL) was stirred in microwave at 150℃ for 1h, cooled, concentrated to dryness, and the crude product (R)-N-(1-() was purified by column chromatography. 3-bromophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (120mg).
MS m/z(ESI):703.1 705.2[M+H] +. MS m/z(ESI): 703.1 705.2[M+H] + .
第二步：(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-N-(1-(3-bromophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indino Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000076
Figure PCTCN2020082042-appb-000076
将(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(120mg)溶于二氯甲烷(2mL)，加三氟乙酸(2mL)，室温搅拌2h，浓缩至干，柱层析纯化得黄色固体产物(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(26mg，二步产率30％)。The (R)-N-(1-(3-bromophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4, 5-f]Indazole-6-carboxamide (120mg) was dissolved in dichloromethane (2mL), added with trifluoroacetic acid (2mL), stirred at room temperature for 2h, concentrated to dryness, purified by column chromatography to obtain a yellow solid product (R) -N-(1-(3-Bromophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide( 26mg, two-step yield 30%).
1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),9.48(d,J＝7.9Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.56–7.47(m,2H),7.31(dq,J＝7.5,1.4Hz,1H),7.16(t,J＝7.5Hz,1H),6.97(d,J＝9.3Hz,1H),5.20(dqt,J＝8.8,6.7,0.9Hz,1H),1.59(d,J＝6.9Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47 (s, 1H), 9.48 (d, J = 7.9 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s,1H),7.96–7.89(m,2H),7.56–7.47(m,2H),7.31(dq,J=7.5,1.4Hz,1H),7.16(t,J=7.5Hz,1H),6.97 (d,J=9.3Hz,1H), 5.20(dqt,J=8.8,6.7,0.9Hz,1H), 1.59(d,J=6.9Hz,3H).
MS m/z(ESI):461.1，463.0[M+H] +. MS m/z(ESI): 461.1, 463.0[M+H] + .
实施例14Example 14
(R)-N-(1-(3-甲氧苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(3-Methoxyphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6 -Formamide preparation
Figure PCTCN2020082042-appb-000077
Figure PCTCN2020082042-appb-000077
第一步：(R)-N-(1-(3-甲氧苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并 [4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-N-(1-(3-methoxyphenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazole Preparation of and [4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000078
Figure PCTCN2020082042-appb-000078
将3-(吡啶-4-基)-6-(三氯甲基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑(100mg，0.17mmol)溶于12mL乙腈和水的混合溶剂中(乙腈：水＝3：1)，加入碳酸氢钠(141mg，1.70mmol)和(R)-(+)-1-(3-甲氧基苯基)乙胺(28mg，0.19mmol)，氮气保护下60℃反应2小时。将反应液浓缩后，加入20mL水，水层用乙酸乙酯萃取(20mL*3)，乙酸乙酯层用饱和氯化钠溶液洗涤、无水硫酸钠干燥后，柱层析(洗脱剂：石油醚：乙酸乙酯＝1：1～乙酸乙酯)，得棕色油状物粗产物(50mg)，直接用于下一步反应。Add 3-(pyridin-4-yl)-6-(trichloromethyl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole (100mg, 0.17mmol) Dissolve in 12 mL of a mixed solvent of acetonitrile and water (acetonitrile: water = 3:1), add sodium bicarbonate (141mg, 1.70mmol) and (R)-(+)-1-(3-methoxyphenyl) Ethylamine (28mg, 0.19mmol) was reacted at 60°C for 2 hours under nitrogen protection. After the reaction solution was concentrated, 20 mL of water was added, the aqueous layer was extracted with ethyl acetate (20 mL*3), the ethyl acetate layer was washed with saturated sodium chloride solution, dried with anhydrous sodium sulfate, and column chromatography (eluent: Petroleum ether: ethyl acetate = 1:1 ~ ethyl acetate) to obtain a brown oily crude product (50 mg), which was directly used in the next reaction.
MS m/z(ESI):655.1[M+H] +. MS m/z(ESI): 655.1[M+H] + .
第二步：(R)-N-(1-(3-甲氧苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-N-(1-(3-methoxyphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f] Preparation of indazole-6-carboxamide
Figure PCTCN2020082042-appb-000079
Figure PCTCN2020082042-appb-000079
将(R)-N-(1-(3-甲氧苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(50mg)溶于4mL二氯甲烷，同时加入4mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥。所得有机溶剂旋干后通过薄层层析(展开剂：二氯甲烷：甲醇＝10：1)纯化，得黄色固体产物(R)-N-(1-(3-甲氧苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(7.1mg，产率10％)。The (R)-N-(1-(3-methoxyphenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4 ,5-f]Indazole-6-carboxamide (50 mg) was dissolved in 4 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added at the same time. After reacting for 3 hours at room temperature, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by thin-layer chromatography (developing solvent: dichloromethane: methanol = 10:1) to obtain the yellow solid product (R)-N-(1-(3-methoxyphenyl)ethyl )-3-(Pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (7.1 mg, yield 10%).
1H NMR(400MHz,DMSO-d 6)δ13.45-13.39(m,1H),13.21-13.08(m,1H),9.43–9.33(m,1H),8.68–8.62(m,2H),8.00–7.94(m,2H),7.70(s,2H),7.35–7.27(m,2H),6.90–6.82(m,2H),5.25–5.21(m,1H),3.81(s,3H),1.47(d,J＝6.8Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.45-13.39 (m, 1H), 13.21-13.08 (m, 1H), 9.43-9.33 (m, 1H), 8.68-8.62 (m, 2H), 8.00 --7.94 (m, 2H), 7.70 (s, 2H), 7.35 - 7.27 (m, 2H), 6.90 - 6.82 (m, 2H), 5.25 - 5.21 (m, 1H), 3.81 (s, 3H), 1.47 (d,J=6.8Hz,3H).
MS m/z(ESI):413.1[M+H] +. MS m/z(ESI): 413.1[M+H] + .
实施例15Example 15
(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(2-Hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-methan Preparation of amide
Figure PCTCN2020082042-appb-000080
Figure PCTCN2020082042-appb-000080
第一步：(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000081
Figure PCTCN2020082042-appb-000081
3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯(100mg,0.18mmol)和(S)-2-氨基-2-苯基乙烷-1-醇(102mg,0.75mmol)溶于二氧六环(3mL)中，150℃微波反应3小时。浓缩后柱层析[洗脱剂:CH 2Cl 2～CH 2Cl 2/MeOH(97:3)]得产品(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(80mg,产率67％)。 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester (100mg, 0.18mmol) and ( S)-2-Amino-2-phenylethane-1-ol (102mg, 0.75mmol) was dissolved in dioxane (3mL) and reacted in microwave at 150°C for 3 hours. After concentration, column chromatography [eluent: CH 2 Cl 2 ～CH 2 Cl 2 /MeOH(97:3)] to obtain the product (S)-N-(2-hydroxy-1-phenylethyl)-3- (Pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (80 mg, 67% yield).
MS m/z(ESI):641.2[M+H] +. MS m/z(ESI):641.2[M+H] + .
第二步：(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole Preparation of -6-formamide
Figure PCTCN2020082042-appb-000082
Figure PCTCN2020082042-appb-000082
往(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(80mg,0.13mmol)的二氯甲烷(2mL)溶液里加三氟乙酸(2mL)，室温搅拌3小时。旋干反应液，加甲醇(5mL)溶解，用氨水调碱，再浓缩后薄层层析[CH 2Cl 2/MeOH(10：1)～CH 2Cl 2/MeOH(8：1)]得产品(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(5.4mg,产率11％)。 To (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5 -f] Indazole-6-carboxamide (80mg, 0.13mmol) in dichloromethane (2mL) was added with trifluoroacetic acid (2mL) and stirred at room temperature for 3 hours. Rotate the reaction solution to dryness, add methanol (5mL) to dissolve, adjust the base with ammonia, and then concentrate and then thin layer chromatography [CH 2 Cl 2 /MeOH (10:1) ~ CH 2 Cl 2 /MeOH (8:1)] to obtain Product (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide (5.4 mg, yield 11%).
1H NMR(400MHz,DMSO)δ13.55-13.01(m,2H),9.25-9.14(m,1H),8.72(d,J＝4Hz,2H),8.50(s,0.6H),8.07-7.91(m,2.7H),7.59(s,0.6H),7.45(d,J＝8Hz,2H),7.35(t,J＝8Hz,2H),7.28-7.23(m,1H),5.14-5.09(m,2H),3.84-3.77(m,2H). 1 H NMR (400MHz, DMSO) δ 13.55-13.01 (m, 2H), 9.25-9.14 (m, 1H), 8.72 (d, J = 4Hz, 2H), 8.50 (s, 0.6H), 8.07-7.91 (m,2.7H),7.59(s,0.6H),7.45(d,J=8Hz,2H),7.35(t,J=8Hz,2H),7.28-7.23(m,1H),5.14-5.09( m,2H),3.84-3.77(m,2H).
MS m/z(ESI):399.1[M+H] +. MS m/z(ESI): 399.1[M+H] + .
实施例16Example 16
(R)-3-(吡啶-4-基)-N-(1-(m-苯甲基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-3-(pyridin-4-yl)-N-(1-(m-benzyl)ethyl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000083
Figure PCTCN2020082042-appb-000083
第一步：(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000084
Figure PCTCN2020082042-appb-000084
3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯(100mg,0.19mmol)和(R)-1-(m-苯甲基)乙烷-1-胺(101mg,0.75mmol)在N-甲基吡咯烷酮(3mL)中150℃微波搅拌3小时。浓缩后柱层析(洗脱剂:CH 2Cl 2～CH 2Cl 2/MeOH(97:3))得产品(R)-3-(吡啶-4-基)-N-(1-(m-苯甲基)乙基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(80mg,产率67％)。 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester (100mg, 0.19mmol) and ( R)-1-(m-benzyl)ethane-1-amine (101 mg, 0.75 mmol) was stirred in N-methylpyrrolidone (3 mL) at 150°C in microwave for 3 hours. After concentration, column chromatography (eluent: CH 2 Cl 2 ～CH 2 Cl 2 /MeOH (97:3)) to obtain the product (R)-3-(pyridin-4-yl)-N-(1-(m -Benzyl)ethyl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (80 mg, yield 67%).
MS m/z(ESI):639.4[M+H] +. MS m/z(ESI): 639.4[M+H] + .
第二步：(R)-3-(吡啶-4-基)-N-(1-(m-苯甲基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-3-(pyridin-4-yl)-N-(1-(m-benzyl)ethyl)-1,7-dihydroimidazo[4,5-f]indole Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000085
Figure PCTCN2020082042-appb-000085
往(R)-3-(吡啶-4-基)-N-(1-(m-苯甲基)乙基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(80mg,0.13mmol)的二氯甲烷(2mL)溶液里加三氟乙酸(2mL)，室温搅拌3小时。旋干反应液，加甲醇(5mL)溶解，用氨水调碱，再浓缩后薄层层析(CH 2Cl 2/MeOH(8：1))得产品(R)-3-(吡啶-4-基)-N-(1-(m-苯甲基)乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(10mg,产率20％)。 To (R)-3-(pyridin-4-yl)-N-(1-(m-benzyl)ethyl)-1-tritylmethyl-1,7-dihydroimidazo[4, 5-f]Indazole-6-carboxamide (80mg, 0.13mmol) in dichloromethane (2mL) was added with trifluoroacetic acid (2mL) and stirred at room temperature for 3 hours. Rotate the reaction solution to dryness, add methanol (5mL) to dissolve, adjust the alkali with ammonia, and then concentrate and thin layer chromatography (CH 2 Cl 2 /MeOH (8:1)) to obtain the product (R)-3-(pyridine-4- Yl)-N-(1-(m-benzyl)ethyl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (10 mg, yield 20%).
1H NMR(400MHz,MeOD)δ8.57(d,J＝4Hz,2H),8.37(s,0.6H),8.15(s,0.4H),8.02(d,J＝4Hz,2H),7.79(s,0.4H),7.56(s,0.6H),7.42-7.30(m,3H),7.12-08(m,1H),5.12-5.16(m,1H),2.22(s,3H),1.52(d,J＝8Hz,3H). 1 H NMR (400MHz, MeOD) δ8.57 (d, J = 4Hz, 2H), 8.37 (s, 0.6H), 8.15 (s, 0.4H), 8.02 (d, J = 4Hz, 2H), 7.79 ( s, 0.4H), 7.56 (s, 0.6H), 7.42-7.30 (m, 3H), 7.12-08 (m, 1H), 5.12-5.16 (m, 1H), 2.22 (s, 3H), 1.52 ( d, J=8Hz, 3H).
MS m/z(ESI):397.2[M+H] +. MS m/z(ESI): 397.2[M+H] + .
实施例17Example 17
(R)-N-(1-(3-氯苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制 备(R)-N-(1-(3-chlorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000086
Figure PCTCN2020082042-appb-000086
第一步：(R)-N-(1-(3-氯苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-N-(1-(3-chlorophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo Preparation of [4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000087
Figure PCTCN2020082042-appb-000087
将3-(吡啶-4-基)-6-(三氯甲基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑(2.0g，3.36mmol)溶于40mL四氢呋喃和水的混合溶剂中(四氢呋喃：水＝3：1)，加入碳酸氢钠(2.82g，33.6mmol)和(R)-1-(3-氯苯基)乙胺(476mg，3.70mmol)，氮气保护下室温反应4小时。将反应液浓缩后，加入40mL水，水层用乙酸乙酯萃取(40mL*3)，乙酸乙酯层用饱和氯化钠溶液洗涤、无水硫酸钠干燥后，柱层析(洗脱剂：石油醚：乙酸乙酯＝1：1～乙酸乙酯)，得棕色固体粗产物(540mg)，直接用于下一步反应。Add 3-(pyridin-4-yl)-6-(trichloromethyl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole (2.0g, 3.36mmol ) Was dissolved in a mixed solvent of 40mL of tetrahydrofuran and water (tetrahydrofuran: water=3:1), sodium bicarbonate (2.82g, 33.6mmol) and (R)-1-(3-chlorophenyl)ethylamine (476mg) were added , 3.70 mmol), react at room temperature for 4 hours under nitrogen protection. After the reaction solution was concentrated, 40 mL of water was added, the aqueous layer was extracted with ethyl acetate (40 mL*3), the ethyl acetate layer was washed with saturated sodium chloride solution, dried with anhydrous sodium sulfate, and column chromatography (eluent: Petroleum ether: ethyl acetate = 1:1 ~ ethyl acetate), a brown solid crude product (540 mg) was obtained, which was directly used in the next reaction.
MS m/z(ESI):659.1[M+H] +. MS m/z(ESI): 659.1[M+H] + .
第二步：(R)-N-(1-(3-氯苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-N-(1-(3-chlorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indino Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000088
Figure PCTCN2020082042-appb-000088
将(R)-N-(1-(3-氯苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(540mg)溶于5mL二氯甲烷，同时加入5mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥。所得有机溶剂旋干后通过柱层析(洗脱剂：二氯甲烷：甲醇＝10：1)纯化，得淡黄色固体产物(R)-N-(1-(3-氯苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(300mg，2步产率21％)。(R)-N-(1-(3-chlorophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4, 5-f]Indazole-6-carboxamide (540 mg) was dissolved in 5 mL of dichloromethane, and 5 mL of trifluoroacetic acid was added at the same time. After 3 hours of reaction at room temperature, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by column chromatography (eluent: dichloromethane: methanol = 10:1) to obtain the light yellow solid product (R)-N-(1-(3-chlorophenyl)ethyl )-3-(Pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (300 mg, 21% yield in 2 steps).
1H NMR(400MHz,DMSO)δ13.48(s,0.4H),13.41(s,0.6H),13.25(s,0.4H),13.11(s,0.6H),9.60(d,J＝8.5Hz,0.4H),9.53(d,J＝8.5Hz,0.6H),8.78–8.67 (m,2H),8.46(s,0.6H),8.13(s,0.4H),8.06(d,J＝5.9Hz,1.2H),7.99(d,J＝5.9Hz,0.8H),7.90(s,0.4H),7.59(s,1.6H),7.46–7.31(m,3H),5.24(p,J＝7.0Hz,1H),1.57(d,J＝7.0Hz,3H). 1 H NMR(400MHz,DMSO)δ13.48(s,0.4H), 13.41(s, 0.6H), 13.25(s, 0.4H), 13.11(s, 0.6H), 9.60(d,J=8.5Hz) ,0.4H),9.53(d,J=8.5Hz,0.6H),8.78–8.67 (m,2H),8.46(s,0.6H),8.13(s,0.4H),8.06(d,J=5.9 Hz,1.2H),7.99(d,J=5.9Hz,0.8H),7.90(s,0.4H),7.59(s,1.6H),7.46-7.31(m,3H), 5.24(p,J= 7.0Hz, 1H), 1.57(d, J=7.0Hz, 3H).
MS m/z(ESI):417.1[M+H] +. MS m/z(ESI): 417.1[M+H] + .
实施例18Example 18
(S)-N-(1-(3-氯苯基)-2-羟基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(1-(3-chlorophenyl)-2-hydroxyethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indino Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000089
Figure PCTCN2020082042-appb-000089
第一步：(S)-N-(1-(3-氯苯基)-2-羟基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(1-(3-chlorophenyl)-2-hydroxyethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7- Preparation of dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000090
Figure PCTCN2020082042-appb-000090
将甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(100mg,0.19mmol)置于微波反应管中，加入(S)-2-氨基-2-(3-氯苯基)乙醇(2.0mL)，加热至150℃反应60min。待冷却至室温后，向反应液中加入20mL乙酸乙酯，乙酸乙酯层用饱和氯化铵溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥后柱层析(洗脱剂：二氯甲烷～二氯甲烷：甲醇＝20：1)纯化，得粗产物(90mg)，直接用于下一步反应。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (100mg, 0.19mmol) Place in a microwave reaction tube, add (S)-2-amino-2-(3-chlorophenyl)ethanol (2.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution, the ethyl acetate layer was washed with saturated ammonium chloride solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate and then column chromatography (eluent: Dichloromethane ~ dichloromethane: methanol = 20:1) to obtain the crude product (90 mg), which was directly used in the next reaction.
MS m/z(ESI):675.1[M+H] +. MS m/z(ESI): 675.1[M+H] + .
第二步：(S)-N-(1-(3-氯苯基)-2-羟基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (S)-N-(1-(3-chlorophenyl)-2-hydroxyethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5 -f] Preparation of indazole-6-carboxamide
Figure PCTCN2020082042-appb-000091
Figure PCTCN2020082042-appb-000091
将(S)-N-(1-(3-氯苯基)-2-羟基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(90mg)溶于4mL二氯甲烷，同时加入4mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥。 所得有机溶剂旋干后通过薄层层析(展开剂：二氯甲烷：甲醇＝10：1)纯化，得黄色固体产物(S)-N-(1-(3-氯苯基)-2-羟基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(22mg，2步产率26％)。(S)-N-(1-(3-chlorophenyl)-2-hydroxyethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazole And [4,5-f] indazole-6-carboxamide (90 mg) was dissolved in 4 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added at the same time. After reacting at room temperature for 3 hours, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by thin layer chromatography (developing solvent: dichloromethane: methanol = 10:1) to obtain the yellow solid product (S)-N-(1-(3-chlorophenyl)-2- Hydroxyethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (22 mg, 26% yield in 2 steps).
1H NMR(400MHz,DMSO-d 6)δ13.49(s,0.4H),13.41(s,0.6H),13.27(s,0.4H),13.14(s,0.6H),9.36(d,J＝8.5Hz,0.4H),9.27(d,J＝8.5Hz,0.6H),8.74–8.71(m,2H),8.50(s,0.6H),8.14(s,0.4H),8.07(d,J＝6.1Hz,1.2H),7.99(d,J＝6.1Hz,0.8H),7.91(s,0.4H),7.60(s,0.6H),7.56(s,1H),7.47–7.28(m,3H),5.19–5.07(m,2H),3.89–3.71(m,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.49 (s, 0.4H), 13.41 (s, 0.6H), 13.27 (s, 0.4H), 13.14 (s, 0.6H), 9.36 (d, J =8.5Hz,0.4H),9.27(d,J=8.5Hz,0.6H),8.74-8.71(m,2H),8.50(s,0.6H),8.14(s,0.4H),8.07(d, J=6.1Hz,1.2H),7.99(d,J=6.1Hz,0.8H),7.91(s,0.4H),7.60(s,0.6H),7.56(s,1H),7.47–7.28(m ,3H), 5.19-5.07(m,2H), 3.89-3.71(m,2H).
MS m/z(ESI):433.1[M+H] +. MS m/z(ESI): 433.1[M+H] + .
实施例19Example 19
(R)-N-(1-(3,5-二甲氧苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(3,5-Dimethoxyphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]in Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000092
Figure PCTCN2020082042-appb-000092
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-(3,5-二甲氧基苯基)乙-1-胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(3,5-二甲氧苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- (3,5-Dimethoxyphenyl)ethyl-1-amine is used as raw material. Refer to Example 2 Steps 6 and 7 to obtain (R)-N-(1-(3,5-Dimethoxyphenyl)ethyl Yl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,CDCl 3)δ8.75(s,2H),8.27(s,1H),7.90(d,J＝8.3Hz,3H),7.65(s,1H),6.65(s,2H),6.36(s,1H),4.97(s,1H),3.79(s,6H),1.48(s,3H). 1 H NMR(400MHz, CDCl 3 )δ8.75(s,2H), 8.27(s,1H), 7.90(d,J=8.3Hz,3H), 7.65(s,1H), 6.65(s,2H) , 6.36(s, 1H), 4.97(s, 1H), 3.79(s, 6H), 1.48(s, 3H).
MS m/z(ESI):443.1[M+H] +. MS m/z(ESI): 443.1[M+H] + .
实施例20Example 20
(S)-N-(2-羟基-1-(m-苯甲基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(2-Hydroxy-1-(m-benzyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]in Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000093
Figure PCTCN2020082042-appb-000093
第一步：(S)-N-(2-羟基-1-(m-苯甲基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(2-hydroxy-1-(m-benzyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7- Preparation of dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000094
Figure PCTCN2020082042-appb-000094
将甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(100mg,0.19mmol)置于微波反应管中，加入(S)-2-氨基-2-(间甲苯基)乙醇(2.0mL)，加热至150℃反应60min。待冷却至室温后，向反应液中加入20mL乙酸乙酯，乙酸乙酯层用饱和氯化铵溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥后柱层析(洗脱剂：二氯甲烷～二氯甲烷：甲醇＝20：1)纯化，得粗产物(75mg)，直接用于下一步反应。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (100mg, 0.19mmol) Place in a microwave reaction tube, add (S)-2-amino-2-(m-tolyl)ethanol (2.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution, the ethyl acetate layer was washed with saturated ammonium chloride solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate and then column chromatography (eluent: Dichloromethane ~ dichloromethane: methanol = 20:1) to obtain a crude product (75 mg), which was directly used in the next reaction.
MS m/z(ESI):655.2[M+H] +. MS m/z(ESI): 655.2[M+H] + .
第二步：(S)-N-(2-羟基-1-(m-苯甲基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (S)-N-(2-hydroxy-1-(m-benzyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5 -f] Preparation of indazole-6-carboxamide
Figure PCTCN2020082042-appb-000095
Figure PCTCN2020082042-appb-000095
将(S)-N-(2-羟基-1-(m-苯甲基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(75mg)溶于4mL二氯甲烷，同时加入4mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥。所得有机溶剂旋干后通过薄层层析(展开剂：二氯甲烷：甲醇＝10：1)纯化，得黄色固体产物(S)-N-(1-(3-氯苯基)-2-羟基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(12mg，2步产率15％)。Add (S)-N-(2-hydroxy-1-(m-benzyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazole And [4,5-f] indazole-6-carboxamide (75 mg) was dissolved in 4 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added at the same time. After reacting for 3 hours at room temperature, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by thin layer chromatography (developing solvent: dichloromethane: methanol = 10:1) to obtain the yellow solid product (S)-N-(1-(3-chlorophenyl)-2- Hydroxyethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (12 mg, 15% yield for 2 steps).
1H NMR(400MHz,DMSO)δ13.49(s,0.4H),13.42(s,0.6H),13.26(s,0.4H),13.13(s,0.6H),9.26–9.08(m,1H),8.73(d,J＝5.5Hz,2H),8.57–7.54(m,4H),7.27–7.21(m,3H),7.08(d,J＝5.7Hz,1H),5.11–5.06(m,2H),3.88–3.79(m,1H),3.78–3.69(m,1H),2.31(s,3H). 1 H NMR (400MHz, DMSO) δ 13.49 (s, 0.4H), 13.42 (s, 0.6H), 13.26 (s, 0.4H), 13.13 (s, 0.6H), 9.26-9.08 (m, 1H) ,8.73(d,J=5.5Hz,2H),8.57–7.54(m,4H),7.27–7.21(m,3H),7.08(d,J=5.7Hz,1H),5.11–5.06(m,2H) ), 3.88–3.79(m,1H), 3.78–3.69(m,1H), 2.31(s,3H).
MS m/z(ESI):413.1[M+H] +. MS m/z(ESI): 413.1[M+H] + .
实施例21Example 21
(R)-N-(1-(3-氰基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(3-cyanophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6 -Formamide preparation
Figure PCTCN2020082042-appb-000096
Figure PCTCN2020082042-appb-000096
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-3-(1-氨基乙基)苯甲腈为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(3-氰基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-3- (1-aminoethyl) benzonitrile is used as raw material. Refer to Example 2 Steps 6 and 7 to obtain (R)-N-(1-(3-cyanophenyl)ethyl)-3-(pyridine-4- Yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),9.48(d,J＝7.9Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.78(q,J＝1.4Hz,1H),7.58(ddq,J＝6.8,3.9,1.5Hz,2H),7.43(t,J＝7.5Hz,1H),6.97(d,J＝9.3Hz,1H),5.20(dddd,J＝9.3,7.9,6.8,5.8Hz,1H),1.59(d,J＝6.8Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47 (s, 1H), 9.48 (d, J = 7.9 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s,1H),7.96–7.89(m,2H),7.78(q,J=1.4Hz,1H),7.58(ddq,J=6.8,3.9,1.5Hz,2H),7.43(t,J=7.5Hz ,1H), 6.97(d,J=9.3Hz,1H), 5.20(dddd,J=9.3,7.9,6.8,5.8Hz,1H), 1.59(d,J=6.8Hz,3H).
MS m/z(ESI):408.2[M+H] +. MS m/z(ESI): 408.2[M+H] + .
实施例22Example 22
N-(2-羟基-1-(3-甲氧苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备N-(2-hydroxy-1-(3-methoxyphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6 -Formamide preparation
Figure PCTCN2020082042-appb-000097
Figure PCTCN2020082042-appb-000097
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和2-氨基-2-(3-甲氧基苯基)乙-1-醇为原料参考实施例2步骤六和步骤七得到N-(2-羟基-1-(3-甲氧苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and 2-amino-2- (3-Methoxyphenyl) ethane-1-alcohol was used as raw material. Refer to Example 2 Steps 6 and 7 to obtain N-(2-hydroxy-1-(3-methoxyphenyl)ethyl)-3-( (Pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
MS m/z(ESI):429.1[M+H] +. MS m/z(ESI): 429.1[M+H] + .
实施例23Example 23
(S)-N-(2-氨基-2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(2-Amino-2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole Preparation of -6-formamide
Figure PCTCN2020082042-appb-000098
Figure PCTCN2020082042-appb-000098
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(S)-2-氨基-2-苯基乙酰胺为原料参考实施例2步骤六和步骤七得到(S)-N-(2-氨基-2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (S)-2- Amino-2-phenylacetamide was used as the raw material. Refer to Example 2 Step 6 and Step 7 to obtain (S)-N-(2-amino-2-carbonyl-1-phenylethyl)-3-(pyridine-4- Yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.48(s,1H),12.72(s,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.67(d,J＝10.8Hz,1H),7.54–7.45(m,2H),7.42–7.32(m,3H),6.97(s,2H),5.67(dt,J＝11.0,1.0Hz,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.48 (s, 1H), 12.72 (s, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 (s, 1H), 7.96–7.89(m,2H), 7.67(d,J=10.8Hz,1H), 7.54–7.45(m,2H), 7.42–7.32(m,3H), 6.97(s,2H), 5.67(dt, J = 11.0, 1.0 Hz, 1H).
MS m/z(ESI):412.1[M+H] +. MS m/z(ESI): 412.1[M+H] + .
实施例24Example 24
(S)-N-(氰基(苯基)甲基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(cyano(phenyl)methyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide preparation
Figure PCTCN2020082042-appb-000099
Figure PCTCN2020082042-appb-000099
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(S)-2-氨基-2-苯基乙腈为原料参考实施例2步骤六和步骤七得到(S)-N-(氰基(苯基)甲基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (S)-2- Amino-2-phenylacetonitrile was used as the raw material to obtain (S)-N-(cyano(phenyl)methyl)-3-(pyridin-4-yl)-1,7- in step 6 and step 7 of Example 2. Dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.47-13.11(m,2H),8.85-8.71(m,3H),8.45(s,0.7H),8.15(s,0.3H),8.08-7.98(m,2H),7.85(s,0.3H),7.51(s,0.7H),7.42-7.33(m,5H),5.88(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47-13.11 (m, 2H), 8.85-8.71 (m, 3H), 8.45 (s, 0.7H), 8.15 (s, 0.3H), 8.08-7.98 (m, 2H), 7.85 (s, 0.3H), 7.51 (s, 0.7H), 7.42-7.33 (m, 5H), 5.88 (m, 1H).
MS m/z(ESI):394.1[M+H] +. MS m/z(ESI): 394.1[M+H] + .
实施例25Example 25
(R)-N-(1-(4-氨基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(4-Aminophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000100
Figure PCTCN2020082042-appb-000100
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-4-(1-氨基乙基)苯胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(4-氨基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-4- (1-aminoethyl)aniline is used as the raw material. Refer to Example 2 in step six and step seven to obtain (R)-N-(1-(4-aminophenyl)ethyl)-3-(pyridin-4-yl)- 1,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.43-13.37(m,1H),13.18-13.03(m,1H),9.43–9.33(m,1H),8.68–8.62(m,2H),8.00–7.94(m,2H),7.70(s,2H),7.19–7.11(m,2H),6.53–6.48(m,2H),5.25–5.21(m,1H),4.33(s,2H),1.55(d,J＝7.0Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.43-13.37 (m, 1H), 13.18-13.03 (m, 1H), 9.43-9.33 (m, 1H), 8.68-8.62 (m, 2H), 8.00 --7.94 (m, 2H), 7.70 (s, 2H), 7.19 - 7.11 (m, 2H), 6.53 - 6.48 (m, 2H), 5.25 - 5.21 (m, 1H), 4.33 (s, 2H), 1.55 (d,J=7.0Hz,3H).
MS m/z(ESI):398.1[M+H] +. MS m/z(ESI): 398.1[M+H] + .
实施例26Example 26
(R)-N-(1-(3,5-二氯苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(3,5-Dichlorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole Preparation of -6-formamide
Figure PCTCN2020082042-appb-000101
Figure PCTCN2020082042-appb-000101
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-(3,5-二氯苯基)乙-1-胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(3,5-二氯苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备方法参照实施例2。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- (3,5-Dichlorophenyl)ethyl-1-amine is used as a raw material. Refer to Example 2 Steps 6 and 7 to obtain (R)-N-(1-(3,5-Dichlorophenyl)ethyl)- Refer to Example 2 for the preparation method of 3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,CDCl 3)δ8.75(s,2H),8.29(s,1H),7.89(d,J＝0.8Hz,3H),7.56(s,1H),7.34(s,2H),7.28(s,1H),4.97(s,1H),1.48(s,3H). 1 H NMR (400MHz, CDCl 3 ) δ8.75 (s, 2H), 8.29 (s, 1H), 7.89 (d, J = 0.8Hz, 3H), 7.56 (s, 1H), 7.34 (s, 2H) , 7.28 (s, 1H), 4.97 (s, 1H), 1.48 (s, 3H).
MS m/z(ESI):451.1，453.1[M+H] +. MS m/z(ESI): 451.1, 453.1[M+H] + .
实施例27Example 27
(R)-N-(1-(3-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(3-Fluorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000102
Figure PCTCN2020082042-appb-000102
第一步：(R)-N-(1-(3-氟苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-N-(1-(3-fluorophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo Preparation of [4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000103
Figure PCTCN2020082042-appb-000103
将甲基3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(200mg,0.37mmol)置于微波反应管中，加入(R)-1-(3-氟苯基)乙烷-1-胺(1.0mL)，加热至150℃反应60min。待冷却至室温后，向反应液中加入20mL乙酸乙酯，乙酸乙酯层用饱和氯化铵溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥后柱层析，得粗产物(R)-N-(1-(3-氟苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(160mg,产率67％)，直接用于下一步反应。Methyl 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (200mg, 0.37mmol) Place in a microwave reaction tube, add (R)-1-(3-fluorophenyl)ethane-1-amine (1.0 mL), and heat to 150° C. to react for 60 min. After cooling to room temperature, 20 mL of ethyl acetate was added to the reaction solution. The ethyl acetate layer was washed with saturated ammonium chloride solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate and column chromatography was used to obtain the crude product ( R)-N-(1-(3-Fluorophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5- f] Indazole-6-carboxamide (160 mg, 67% yield), used directly in the next reaction.
MS m/z(ESI):643.3[M+H] +. MS m/z(ESI):643.3[M+H] + .
第二步：(R)-N-(1-(3-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (R)-N-(1-(3-fluorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indole Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000104
Figure PCTCN2020082042-appb-000104
将(R)-N-(1-(3-氟苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺粗品(160mg)溶于4mL二氯甲烷，同时加入4mL三氟乙酸，室温反应3小时后，将反应液旋干。将所得粗品溶于乙酸乙酯和四氢呋喃的混合溶液，用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，之后用无水硫酸钠干燥。所得有机溶剂旋干后通过制备色谱纯化，得黄色固体产物(R)-N-(1-(3-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(30mg,产率27％)。(R)-N-(1-(3-Fluorophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4, 5-f] Indazole-6-carboxamide crude product (160 mg) was dissolved in 4 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added at the same time. After reacting for 3 hours at room temperature, the reaction solution was spin-dried. The obtained crude product was dissolved in a mixed solution of ethyl acetate and tetrahydrofuran, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, and then dried with anhydrous sodium sulfate. The obtained organic solvent was spin-dried and purified by preparative chromatography to obtain the yellow solid product (R)-N-(1-(3-fluorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-di Hydroimidazo[4,5-f]indazole-6-carboxamide (30mg, 27% yield).
1H NMR(400MHz,DMSO-d 6)δ13.46(s,1H),9.50(d,J＝8Hz,1H),8.72(d,J＝8Hz,2H),8.36(s,2H),8.02(d,J＝4Hz,2H),7.62-7.73(m,1H),7.42-7.30(m,2H),7.14-7.06(m,2H),5.27-5.23(m,1H),1.56(d,J＝8Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.46 (s, 1H), 9.50 (d, J = 8 Hz, 1H), 8.72 (d, J = 8 Hz, 2H), 8.36 (s, 2H), 8.02 (d,J=4Hz,2H),7.62-7.73(m,1H),7.42-7.30(m,2H),7.14-7.06(m,2H),5.27-5.23(m,1H),1.56(d, J=8Hz, 3H).
MS m/z(ESI):401.2[M+H] +. MS m/z(ESI): 401.2[M+H] + .
实施例28Example 28
(R)-N-(1-(2-氯苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(2-Chlorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000105
Figure PCTCN2020082042-appb-000105
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-(2-氯苯基)乙-1-胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(2-氯苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- (2-Chlorophenyl) ethyl-1-amine is used as the raw material. Refer to Example 2 Step 6 and Step 7 to obtain (R)-N-(1-(2-chlorophenyl)ethyl)-3-(pyridine-4 -Yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),9.48(d,J＝7.9Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.44–7.36(m,1H),7.36–7.28(m,1H),7.24–7.13(m,2H),6.83(d,J＝9.5Hz,1H),5.28(dqd,J＝9.5,6.8,1.0Hz,1H),1.68(d,J＝6.9Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47 (s, 1H), 9.48 (d, J = 7.9 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s,1H),7.96–7.89(m,2H),7.44–7.36(m,1H),7.36–7.28(m,1H),7.24–7.13(m,2H),6.83(d,J=9.5Hz, 1H), 5.28(dqd,J=9.5,6.8,1.0Hz,1H),1.68(d,J=6.9Hz,3H).
MS m/z(ESI):417.1，419.1[M+H] +. MS m/z(ESI): 417.1, 419.1[M+H] + .
实施例29Example 29
(S)-N-(2-(甲基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰 胺的制备(S)-N-(2-(methylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole Preparation of -6-formamide
Figure PCTCN2020082042-appb-000106
Figure PCTCN2020082042-appb-000106
第一步：(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000107
Figure PCTCN2020082042-appb-000107
3-(吡啶-4-基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯(50mg，0.93mmol)、(S)-2-氨基-2-苯基乙烷-1-醇(640mg，4.66mmol)和NMP(10mL)加入150mL微波中加热搅拌2h，冷却，浓缩至干，柱层析纯化得棕色固体产物(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(340mg，产率56％)。3-(pyridin-4-yl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester (50mg, 0.93mmol), (S )-2-Amino-2-phenylethane-1-ol (640mg, 4.66mmol) and NMP (10mL) were added to 150mL microwave and heated and stirred for 2h, cooled, concentrated to dryness, purified by column chromatography to obtain a brown solid product ( S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f ] Indazole-6-carboxamide (340 mg, yield 56%).
MS m/z(ESI):641.2[M-H] +. MS m/z(ESI):641.2[MH] + .
第二步：(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000108
Figure PCTCN2020082042-appb-000108
(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(220mg，0.34mmol)溶于20mL DMSO，加入IBX(769mg，2.74mmol)，室温搅拌过夜。浓缩至干，柱层析纯化得粗品棕色固体产物(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(300mg)。(S)-N-(2-Hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5- f] Indazole-6-carboxamide (220 mg, 0.34 mmol) was dissolved in 20 mL DMSO, IBX (769 mg, 2.74 mmol) was added, and the mixture was stirred at room temperature overnight. Concentrate to dryness, and purify by column chromatography to obtain crude brown solid product (S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl- 1,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide (300mg).
MS m/z(ESI):639.2[M-H] +. MS m/z(ESI): 639.2[MH] + .
第三步：(S)-N-(2-(甲基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The third step: (S)-N-(2-(methylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-di Preparation of Hydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000109
Figure PCTCN2020082042-appb-000109
(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(100mg)、30％甲胺/甲醇溶液(3mL)和冰醋酸(2mL)溶于二氯甲烷/甲醇(10mL/10mL)中，室温搅拌30min，加入氰基硼氢化钠(49mg，0.78mmol)，室温搅拌过夜。浓缩至干，加水，二氯甲烷提取，干燥，过滤，浓缩至干。柱层析纯化得粘稠物粗品(S)-N-(2-(甲基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(100mg)。(S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5- f] Indazole-6-carboxamide (100mg), 30% methylamine/methanol solution (3mL) and glacial acetic acid (2mL) were dissolved in dichloromethane/methanol (10mL/10mL), stirred at room temperature for 30 min, and added cyano Sodium borohydride (49 mg, 0.78 mmol) was stirred at room temperature overnight. Concentrate to dryness, add water, extract with dichloromethane, dry, filter, and concentrate to dryness. Purified by column chromatography to obtain the crude viscous product (S)-N-(2-(methylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl- 1,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide (100 mg).
MS m/z(ESI):654.2[M-H] +. MS m/z(ESI): 654.2[MH] + .
第四步：(S)-N-(2-(甲基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The fourth step: (S)-N-(2-(methylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5- f] Preparation of indazole-6-carboxamide
Figure PCTCN2020082042-appb-000110
Figure PCTCN2020082042-appb-000110
将上述第三步得到的粘稠物(S)-N-(2-(甲基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(粗品，100mg)溶于二氯甲烷(1mL)，加入三氟乙酸(2mL)，室温搅拌2h。浓缩至干，柱层析纯化得固体产物(S)-N-(2-(甲基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(3.6mg，三步收率7.6％)。Add the viscous (S)-N-(2-(methylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1-trityl group obtained in the third step above -1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (crude product, 100mg) was dissolved in dichloromethane (1mL), trifluoroacetic acid (2mL) was added, and the mixture was stirred at room temperature for 2h. Concentrate to dryness, and purify by column chromatography to obtain the solid product (S)-N-(2-(methylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydro Imidazo[4,5-f]indazole-6-carboxamide (3.6 mg, yield 7.6% in three steps).
MS m/z(ESI):412.1[M+H] +. MS m/z(ESI): 412.1[M+H] + .
实施例30Example 30
(S)-N-(2-(环丙基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(2-(Cyclopropylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indino Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000111
Figure PCTCN2020082042-appb-000111
第一步：(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000112
Figure PCTCN2020082042-appb-000112
3-(吡啶-4-基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯(2g，3.73mmol)、(S)-2-氨基-2-苯基乙烷-1-醇(2.56g，18.66mmol)140℃微波搅拌1h，加水，乙酸乙酯提取，干燥浓缩至干，柱分离纯化得(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(1.46g，产率61％)。3-(pyridin-4-yl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester (2g, 3.73mmol), (S )-2-Amino-2-phenylethane-1-ol (2.56g, 18.66mmol) microwave stirring at 140℃ for 1h, add water, extract with ethyl acetate, dry and concentrate to dryness, column separation and purification to obtain (S)-N -(2-Hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole- 6-formamide (1.46 g, 61% yield).
MS m/z(ESI):641.2[M+H] +. MS m/z(ESI):641.2[M+H] + .
第二步：(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000113
Figure PCTCN2020082042-appb-000113
将(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(1.46g，2.28mmol)溶于DMSO(10mL)，加IBX(5.1g，18.21mmol)，氮气保护下室温搅拌3h。加二氯甲烷，碳酸钠溶液洗，干燥浓缩至干，柱分离纯化得油状物粗品(2.52g)。The (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5 -f] Indazole-6-carboxamide (1.46g, 2.28mmol) was dissolved in DMSO (10mL), added IBX (5.1g, 18.21mmol), and stirred at room temperature for 3h under nitrogen protection. Add dichloromethane, wash with sodium carbonate solution, dry and concentrate to dryness, column separation and purification to obtain crude oil (2.52g).
MS m/z(ESI):639.2[M+H] +. MS m/z(ESI): 639.2[M+H] + .
第三步：(S)-N-(2-(环丙基氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The third step: (S)-N-(2-(cyclopropylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7- Preparation of dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000114
Figure PCTCN2020082042-appb-000114
(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(20mg，0.031mmol)、环丙胺(18mg，0.31mmol)于二氯甲烷/甲醇(10mL/10mL)中搅拌，加醋酸1mL，搅拌30min。加氰基硼氢化钠(10mg，0.16mmol)，搅拌过夜，加水，二氯甲烷提取，干燥浓缩至干，柱分离纯化得油状物，直接用于下步反应。(S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5- f] Indazole-6-carboxamide (20 mg, 0.031 mmol) and cyclopropylamine (18 mg, 0.31 mmol) were stirred in dichloromethane/methanol (10 mL/10 mL), and 1 mL of acetic acid was added and stirred for 30 min. Add sodium cyanoborohydride (10 mg, 0.16 mmol), stir overnight, add water, extract with dichloromethane, dry and concentrate to dryness, column separation and purification to obtain an oily substance, which is directly used in the next reaction.
MS m/z(ESI):704.2[M+H] +. MS m/z(ESI): 704.2[M+H] + .
第四步：(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The fourth step: (S)-N-(2-((2,2-difluoroethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-di Preparation of Hydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000115
Figure PCTCN2020082042-appb-000115
向上步得到的油状物溶于二氯甲烷(2mL)，加入三氟乙酸(2mL)，室温搅拌2h。浓缩至干，柱分离纯化得类白色固体(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(3.5mg，两步产率25％)。The oil obtained in the previous step was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, and the mixture was stirred at room temperature for 2 h. Concentrate to dryness, column separation and purification to obtain off-white solid (S)-N-(2-((2,2-difluoroethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl) )-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (3.5 mg, two-step yield 25%).
1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),9.48(d,J＝7.9Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.40–7.23(m,5H),7.10(d,J＝10.3Hz,1H),5.12(dtt,J＝10.2,6.9,1.0Hz,1H),3.22(dt,J＝7.7,6.1Hz,1H),3.11(ddd,J＝12.8,7.0,6.0Hz,1H),3.00(ddd,J＝12.5,7.0,6.1Hz,1H),2.54–2.41(m,1H),1.05–0.77(m,4H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47 (s, 1H), 9.48 (d, J = 7.9 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s, 1H), 7.96–7.89 (m, 2H), 7.40–7.23 (m, 5H), 7.10 (d, J = 10.3 Hz, 1H), 5.12 (dtt, J = 10.2, 6.9, 1.0 Hz, 1H) ,3.22(dt,J＝7.7,6.1Hz,1H), 3.11(ddd,J＝12.8,7.0,6.0Hz,1H), 3.00(ddd,J＝12.5,7.0,6.1Hz,1H), 2.54–2.41 (m,1H),1.05-0.77(m,4H).
MS m/z(ESI):438.2[M+H] +. MS m/z(ESI): 438.2[M+H] + .
实施例31Example 31
N-(2-氟-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Preparation of N-(2-fluoro-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000116
Figure PCTCN2020082042-appb-000116
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和2-氟-1-苯基乙-1-胺为原料参考实施例2步骤六和步骤七得到N-(2-氟-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and 2-fluoro-1- Phenylethyl-1-amine is used as the raw material. Refer to Example 2 Step 6 and Step 7 to obtain N-(2-fluoro-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydro Imidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),9.48(d,J＝7.9Hz,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.65(d,J＝11.0Hz,1H),7.41–7.32(m,4H),7.32–7.23(m,1H),5.55–5.38(m,1H),4.99(d,J＝7.0Hz,1H),4.87(d,J＝7.1Hz,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.47 (s, 1H), 9.48 (d, J = 7.9 Hz, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 ( s,1H),7.96-7.89(m,2H),7.65(d,J=11.0Hz,1H),7.41-7.32(m,4H),7.32-7.23(m,1H),5.55-5.38(m, 1H), 4.99(d,J=7.0Hz,1H), 4.87(d,J=7.1Hz,1H).
MS m/z(ESI):401.1[M+H] +. MS m/z(ESI): 401.1[M+H] + .
实施例32Example 32
3-(吡啶-4-基)-N-(2,2,2-三氟-1-苯基乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰 胺的制备3-(pyridin-4-yl)-N-(2,2,2-trifluoro-1-phenylethyl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000117
Figure PCTCN2020082042-appb-000117
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和2,2,2-三氟-1-苯基乙-1-胺为原料参考实施例2步骤六和步骤七得到3-(吡啶-4-基)-N-(2,2,2-三氟-1-苯基乙基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and 2,2,2- Trifluoro-1-phenylethyl-1-amine was used as a raw material. Refer to Example 2 Step 6 and Step 7 to obtain 3-(pyridin-4-yl)-N-(2,2,2-trifluoro-1-phenyl) Ethyl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ:13.56(s,1H),13.32(s,1H),9.39(d,J＝7.5Hz,1H),8.68–8.62(m,2H),8.00–7.94(m,2H),7.70(s,2H),7.41–7.30(m,4H),7.30–7.26(m,1H),7.05–6.91(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ: 13.56 (s, 1H), 13.32 (s, 1H), 9.39 (d, J = 7.5 Hz, 1H), 8.68–8.62 (m, 2H), 8.00– 7.94(m,2H), 7.70(s,2H), 7.41--7.30(m,4H), 7.30--7.26(m,1H), 7.05--6.91(m,1H).
MS m/z(ESI):437.1[M+H] +. MS m/z(ESI): 437.1[M+H] + .
实施例33Example 33
(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-b]吡唑并[4,3-e]吡啶-6-甲酰胺的制备(R)-N-(1-Phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-b]pyrazolo[4,3-e] Preparation of pyridine-6-carboxamide
Figure PCTCN2020082042-appb-000118
Figure PCTCN2020082042-appb-000118
第一步：(2,6-二氟吡啶-3-基)(吡啶-4-基)甲醇的制备Step 1: Preparation of (2,6-difluoropyridin-3-yl)(pyridin-4-yl)methanol
Figure PCTCN2020082042-appb-000119
Figure PCTCN2020082042-appb-000119
将2,6-二氟吡啶(3.45g，30mmol)溶于100mL无水四氢呋喃，冷却至-78℃，氮气保护下滴加2.0M LDA的THF/n-heptane溶液(15.8mL，31.5mmol)，之后注射加入4-吡啶甲醛(3.86g，36mmol)。反应由-78℃逐渐升温至室温，检测反应完毕后向反应液中加入1.8mL醋酸，室温下搅拌30分钟。向反应液中直接加入硅胶，旋干后柱层析，得淡黄色固体(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-b]吡唑并[4,3-e]吡啶-6-甲酰胺(5.0g，产率75％)。Dissolve 2,6-difluoropyridine (3.45g, 30mmol) in 100mL of anhydrous tetrahydrofuran, cool to -78°C, add 2.0M LDA in THF/n-heptane solution (15.8mL, 31.5mmol) dropwise under nitrogen protection, Then 4-pyridinecarbaldehyde (3.86 g, 36 mmol) was added by injection. The reaction was gradually warmed from -78°C to room temperature. After the reaction was detected, 1.8 mL of acetic acid was added to the reaction solution and stirred at room temperature for 30 minutes. Silica gel was directly added to the reaction solution, spin-dried, and column chromatography was used to obtain a pale yellow solid (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydro Imidazo[4,5-b]pyrazolo[4,3-e]pyridine-6-carboxamide (5.0 g, yield 75%).
MS m/z(ESI):223.1[M+H] +. MS m/z(ESI): 223.1[M+H] + .
第二步：(2,6-二氟吡啶-3-基)(吡啶-4-基)甲酮的制备Step 2: Preparation of (2,6-difluoropyridin-3-yl)(pyridin-4-yl)methanone
Figure PCTCN2020082042-appb-000120
Figure PCTCN2020082042-appb-000120
将(2,6-二氟吡啶-3-基)(吡啶-4-基)甲醇(5.0g，22.5mmol)溶于250mL DMF，分批次加入PDC(25.4g，67.5mmol)，于室温下反应4小时。将反应液浓缩后，加入200mL水，用乙酸乙酯萃取。乙酸乙酯层用饱和NaCl溶液洗涤，无水硫酸钠干燥后，柱层析，得白色固体(2,6-二氟吡啶-3-基)(吡啶-4-基)甲酮(3.6g，产率73％)。Dissolve (2,6-difluoropyridin-3-yl)(pyridin-4-yl)methanol (5.0g, 22.5mmol) in 250mL DMF, add PDC (25.4g, 67.5mmol) in batches, at room temperature React for 4 hours. After the reaction solution was concentrated, 200 mL of water was added, and extraction was performed with ethyl acetate. The ethyl acetate layer was washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and column chromatography was used to obtain white solid (2,6-difluoropyridin-3-yl)(pyridin-4-yl)methanone (3.6g, The yield is 73%).
MS m/z(ESI):221.1[M+H] +. MS m/z(ESI): 221.1[M+H] + .
第三步：6-氟-3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶的制备The third step: Preparation of 6-fluoro-3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine
Figure PCTCN2020082042-appb-000121
Figure PCTCN2020082042-appb-000121
将(2,6-二氟吡啶-3-基)(吡啶-4-基)甲酮(3.6g，16.4mmol)溶于30mL 1,4-二氧六环，加入85％的水合肼溶液(1.2g，19.7mmol)，室温反应3小时。向反应液中加入硅胶，直接旋干后柱层析，得白色固体6-氟-3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶(2.6g，产率74％)。Dissolve (2,6-difluoropyridin-3-yl)(pyridin-4-yl)methanone (3.6g, 16.4mmol) in 30mL 1,4-dioxane, and add 85% hydrazine hydrate solution ( 1.2g, 19.7mmol), react at room temperature for 3 hours. Silica gel was added to the reaction solution, and the column chromatography was directly spin-dried to obtain a white solid 6-fluoro-3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine (2.6g, product Rate 74%).
MS m/z(ESI):215.1[M+H] +. MS m/z(ESI): 215.1[M+H] + .
第四步：3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-6-胺的制备Step 4: Preparation of 3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-6-amine
Figure PCTCN2020082042-appb-000122
Figure PCTCN2020082042-appb-000122
将6-氟-3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶(0.5g，2.33mmol)置于微波反应管中，加入4mL DMSO以及8mL浓氨水。微波加热至110℃反应2小时。待反应液冷却后倒入100mL水中搅拌，将析出的固体抽滤，滤饼用水洗涤后真空干燥得粗产物3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-6-胺(450mg)。Place 6-fluoro-3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine (0.5g, 2.33mmol) in a microwave reaction tube, add 4mL DMSO and 8mL concentrated ammonia. Microwave heating to 110°C for 2 hours. After the reaction solution is cooled, it is poured into 100 mL of water and stirred. The precipitated solid is filtered off with suction. The filter cake is washed with water and dried in vacuo to obtain the crude product 3-(pyridin-4-yl)-1H-pyrazolo[3,4-b] Pyridine-6-amine (450 mg).
MS m/z(ESI):212.1[M+H] +. MS m/z(ESI): 212.1[M+H] + .
第五步：5-硝基-3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-6-胺的制备Step 5: Preparation of 5-nitro-3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-6-amine
Figure PCTCN2020082042-appb-000123
Figure PCTCN2020082042-appb-000123
将3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-6-胺(450mg，2.13mmol)溶于7.5mL浓硫酸，置于冰浴下，滴加浓硝酸(310mg，3.20mmol)。之后将反应加热至55℃反应6小时。待反应液冷却后，将其倒入50mL冰水中搅拌，用4N NaOH溶液调节pH至中性。将析出的固体抽滤，滤饼用水洗涤后真空干燥，得黄色固体5-硝基-3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-6-胺(200mg，产率36％)。Dissolve 3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-6-amine (450mg, 2.13mmol) in 7.5mL concentrated sulfuric acid, place it in an ice bath, and add the concentrated Nitric acid (310 mg, 3.20 mmol). The reaction was then heated to 55°C for 6 hours. After the reaction solution is cooled, it is poured into 50 mL of ice water and stirred, and the pH is adjusted to neutral with 4N NaOH solution. The precipitated solid was suction filtered, the filter cake was washed with water and dried in vacuo to obtain a yellow solid 5-nitro-3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-6-amine (200mg, yield 36%).
MS m/z(ESI):257.1[M+H] +. MS m/z(ESI): 257.1[M+H] + .
第六步：3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-5,6-二胺的制备Step 6: Preparation of 3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-5,6-diamine
Figure PCTCN2020082042-appb-000124
Figure PCTCN2020082042-appb-000124
将5-硝基-3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-6-胺(200mg，0.78mmol)溶于DMF与THF的混合溶剂中(25mL：25mL)，加入50mg 20％Pd(OH) 2/C(约含50％水)，用氢气球置换数次之后，于室温反应过夜。反应液过滤除去Pd(OH) 2/C，旋干后得粗产物3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-5,6-二胺(200mg)，直接用于下一步反应。 5-nitro-3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-6-amine (200mg, 0.78mmol) was dissolved in a mixed solvent of DMF and THF (25mL ：25mL), add 50mg 20% Pd(OH) 2 /C (contains about 50% water), replace with hydrogen balloon several times, and react overnight at room temperature. The reaction solution was filtered to remove Pd(OH) 2 /C, and the crude product 3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-5,6-diamine (200mg ), directly used in the next reaction.
MS m/z(ESI):227.1[M+H] +。 MS m/z (ESI): 227.1 [M+H] + .
第七步：甲基3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-b]吡唑并[4,3-e]吡啶-6-羧酸酯的制备Step 7: Preparation of methyl 3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-b]pyrazolo[4,3-e]pyridine-6-carboxylate
Figure PCTCN2020082042-appb-000125
Figure PCTCN2020082042-appb-000125
将3-(吡啶-4-基)-1H-吡唑并[3,4-b]吡啶-5,6-二胺(150mg，0.66mmol)溶于10mL甲醇，置于微波反应管中，加入2mL三乙胺以及1.5mL二氯甲氧基乙酸甲酯，微波加热至100℃反应1.5小时。反应液冷却至室温后加入硅胶旋干，柱层析得粗产物甲基3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-b]吡唑并[4,3-e]吡啶-6-羧酸酯(120mg)。Dissolve 3-(pyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-5,6-diamine (150mg, 0.66mmol) in 10mL methanol, place it in a microwave reaction tube, and add 2mL of triethylamine and 1.5mL of methyl dichloromethoxyacetate were heated to 100°C for 1.5 hours in a microwave. After the reaction solution was cooled to room temperature, silica gel was added and spin-dried, and the crude product methyl 3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-b]pyrazolo[4, 3-e] Pyridine-6-carboxylate (120 mg).
MS m/z(ESI):295.1[M+H] +. MS m/z(ESI): 295.1[M+H] + .
第八步：(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-b]吡唑并[4,3-e]吡啶-6-甲酰胺的制备The eighth step: (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-b]pyrazolo[4, 3-e] Preparation of pyridine-6-carboxamide
Figure PCTCN2020082042-appb-000126
Figure PCTCN2020082042-appb-000126
将甲基3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-b]吡唑并[4,3-e]吡啶-6-羧酸酯(130mg，0.44mmol)置于微波反应管中，加入2mL(R)-(+)-1-苯基乙胺，微波加热至140℃反应1小时。待反应液冷却后加入20mL 2-甲基四氢呋喃，有机层先后用饱和NH 4Cl溶液以及饱和NaCl溶液洗涤，无水硫酸钠干燥后，通过制备HPLC得淡黄色固体(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-b]吡唑并[4,3-e]吡啶-6-甲酰胺(6.8mg，产率4％)。 The methyl 3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-b]pyrazolo[4,3-e]pyridine-6-carboxylate (130mg, 0.44mmol ) Place in a microwave reaction tube, add 2mL (R)-(+)-1-phenylethylamine, microwave heating to 140°C for 1 hour. After the reaction solution was cooled, 20mL 2-methyltetrahydrofuran was added. The organic layer was washed with saturated NH 4 Cl solution and saturated NaCl solution successively. After drying with anhydrous sodium sulfate, a pale yellow solid (R)-N-(1) was obtained by preparative HPLC. -Phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-b]pyrazolo[4,3-e]pyridine-6-carboxamide (6.8 mg, yield 4%).
1H NMR(400MHz,DMSO-d 6)δ13.35-113.13(m,2H),9.45-9.34(m,1H),8.72(d,J＝8Hz,2H),8.02(d,J＝8Hz,2H),7.86(s,0.4H),7.57(s,0.6H),7.46-7.23(m,5H),5.24-5.21(m,1H),1.56(d,J＝8Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.35-113.13 (m, 2H), 9.45-9.34 (m, 1H), 8.72 (d, J = 8 Hz, 2H), 8.02 (d, J = 8 Hz, 2H), 7.86 (s, 0.4H), 7.57 (s, 0.6H), 7.46-7.23 (m, 5H), 5.24-5.21 (m, 1H), 1.56 (d, J = 8Hz, 3H).
MS m/z(ESI):384.1[M+H] +. MS m/z(ESI): 384.1[M+H] + .
实施例34Example 34
(R)-N-(1-(2-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(2-Fluorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide preparation
Figure PCTCN2020082042-appb-000127
Figure PCTCN2020082042-appb-000127
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-(2-氟苯基)乙-1-胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(2-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- (2-Fluorophenyl) ethyl-1-amine is used as the raw material. Refer to Example 2 Step 6 and Step 7 to obtain (R)-N-(1-(2-Fluorophenyl)ethyl)-3-(pyridine-4 -Yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.49(s,1H),9.47(d,J＝7.9Hz,1H),8.72(d,J＝5.2Hz,2H),8.35(s,4H),8.03(d,J＝5.1Hz,2H),7.72(s,1H),7.61(t,J＝7.2Hz,1H),7.32(dd,J＝13.2,5.8Hz,1H),7.26–7.14(m,2H),5.57–5.43(m,1H),1.56(d,J＝7.0Hz,3H). 1 H NMR(400MHz,DMSO-d 6 )δ13.49(s,1H), 9.47(d,J=7.9Hz,1H), 8.72(d,J=5.2Hz,2H), 8.35(s,4H) ,8.03(d,J=5.1Hz,2H),7.72(s,1H),7.61(t,J=7.2Hz,1H),7.32(dd,J=13.2,5.8Hz,1H),7.26-7.14( m, 2H), 5.57-5.43 (m, 1H), 1.56 (d, J = 7.0 Hz, 3H).
MS m/z(ESI):401.1[M+H] +. MS m/z(ESI): 401.1[M+H] + .
实施例35Example 35
(R)-N-(1-(3-氯-4-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(R)-N-(1-(3-chloro-4-fluorophenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indino Preparation of azole-6-carboxamide
Figure PCTCN2020082042-appb-000128
Figure PCTCN2020082042-appb-000128
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和(R)-1-(3-氯-4-氟苯基)乙-1-胺为原料参考实施例2步骤六和步骤七得到(R)-N-(1-(3-氯-4-氟苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and (R)-1- (3-Chloro-4-fluorophenyl)ethyl-1-amine is used as raw material. Refer to Example 2 Steps 6 and 7 to obtain (R)-N-(1-(3-chloro-4-fluorophenyl)ethyl )-3-(Pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.42(s,1H),9.46(d,J＝8.4Hz,1H),8.65(d,J＝4.9Hz,2H),8.27(s,5H),7.96(d,J＝5.3Hz,2H),7.66(dd,J＝7.2,1.9Hz,2H),7.48–7.38(m,1H),7.29(ddd,J＝25.1,12.7,7.0Hz,2H),5.23–5.11(m,1H),1.49(d,J＝7.0Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.42 (s, 1H), 9.46 (d, J = 8.4 Hz, 1H), 8.65 (d, J = 4.9 Hz, 2H), 8.27 (s, 5H) ,7.96(d,J=5.3Hz,2H),7.66(dd,J=7.2,1.9Hz,2H),7.48–7.38(m,1H),7.29(ddd,J=25.1,12.7,7.0Hz,2H ), 5.23–5.11 (m, 1H), 1.49 (d, J=7.0Hz, 3H).
MS m/z(ESI):435.1，437.1[M+H] +. MS m/z(ESI): 435.1, 437.1[M+H] + .
实施例36Example 36
N-(3-羟基-1-苯基丙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Preparation of N-(3-hydroxy-1-phenylpropyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000129
Figure PCTCN2020082042-appb-000129
以3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯和3-氨基-3-苯基丙-1-醇为原料参考实施例2步骤六和步骤七得到N-(3-羟基-1-苯基丙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺。With 3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester and 3-amino-3- Phenylpropan-1-ol was used as the raw material. Refer to Example 2 Step 6 and Step 7 to obtain N-(3-hydroxy-1-phenylpropyl)-3-(pyridin-4-yl)-1,7-dihydro Imidazo[4,5-f]indazole-6-carboxamide.
1H NMR(400MHz,DMSO-d 6)δ13.56-13.03(m,2H),9.25-9.14(m,1H),8.70(d,J＝4Hz,2H),8.49(s,0.6H),8.06-7.90(m,2.7H),7.57(s,0.6H),7.46(d,J＝8Hz,2H),7.35(t,J＝8Hz,2H),7.28-7.23(m,1H),5.14-5.09(m,1H),4.50(s,1H),3.85-3.80(m,2H),2.14-2.03(m,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.56-13.03 (m, 2H), 9.25-9.14 (m, 1H), 8.70 (d, J = 4Hz, 2H), 8.49 (s, 0.6H), 8.06-7.90(m,2.7H),7.57(s,0.6H),7.46(d,J=8Hz,2H),7.35(t,J=8Hz,2H),7.28-7.23(m,1H),5.14 -5.09(m,1H),4.50(s,1H),3.85-3.80(m,2H),2.14-2.03(m,2H).
MS m/z(ESI):413.1[M+H] +. MS m/z(ESI): 413.1[M+H] + .
实施例37Example 37
(R)-N-(1-(3-环丙基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(R)-N-(1-(3-Cyclopropylphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole- 6-formamide
Figure PCTCN2020082042-appb-000130
Figure PCTCN2020082042-appb-000130
第一步：(R)-(1-(3-溴苯基)乙基)氨基甲酸叔丁酯的制备The first step: preparation of (R)-(1-(3-bromophenyl)ethyl) t-butyl carbamate
Figure PCTCN2020082042-appb-000131
Figure PCTCN2020082042-appb-000131
往(R)-1-(3-溴苯基)乙胺(500mg，2.5mmol)的二氯甲烷(20mL)溶液里加三乙胺(500mg，5mmol)，0℃滴加二叔丁基二碳酸酯(648mg，3mmol)的二氯甲烷(5mL)溶液，加完搅拌3小时。往反应液加二氯甲烷(50mL)，用饱和柠檬酸水溶液(30mL*2)洗涤，然后用饱和碳酸氢钠水溶液(30mL)洗涤，浓缩有机相得到无色透明油状产物(R)-(1-(3-溴苯基)乙基)氨基甲酸叔丁酯(750mg,产率100％)。To (R)-1-(3-bromophenyl)ethylamine (500mg, 2.5mmol) in dichloromethane (20mL), add triethylamine (500mg, 5mmol), add di-tert-butyl dicarbonate dropwise at 0°C A solution of the ester (648mg, 3mmol) in dichloromethane (5mL) was added and stirred for 3 hours. Dichloromethane (50mL) was added to the reaction solution, washed with saturated citric acid aqueous solution (30mL*2), and then with saturated sodium bicarbonate aqueous solution (30mL). The organic phase was concentrated to obtain a colorless transparent oily product (R)-(1 -(3-Bromophenyl)ethyl) t-butyl carbamate (750 mg, yield 100%).
第二步：(R)-(1-(3-环丙基苯基)乙基)氨基甲酸叔丁酯的制备Step 2: Preparation of tert-butyl (R)-(1-(3-cyclopropylphenyl)ethyl)carbamate
Figure PCTCN2020082042-appb-000132
Figure PCTCN2020082042-appb-000132
(R)-(1-(3-溴苯基)乙基)氨基甲酸叔丁酯(720mg，2.4mmol)，环丙基硼酸(413mg，4.8mmol)，[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(98mg，0.12mmol)和碳酸钾(994mg，7.2mmol)在二氧六环(10mL)和水(1mL)中微波120℃反应1.5小时。反应完，浓缩反应液后柱层析纯化[洗脱剂:石油醚～石油醚/乙酸乙酯(95/5)]得到无色油状产物(R)-(1-(3-环丙基苯基)乙基)氨基甲酸叔丁酯(430mg，产率69％)。(R)-(1-(3-Bromophenyl)ethyl) tert-butyl carbamate (720mg, 2.4mmol), cyclopropylboronic acid (413mg, 4.8mmol), [1,1'-bis(diphenyl) Phosphine) ferrocene] dichloride palladium dichloromethane complex (98mg, 0.12mmol) and potassium carbonate (994mg, 7.2mmol) were reacted in dioxane (10mL) and water (1mL) in microwave at 120℃ 1.5 hours. After the reaction, the reaction solution is concentrated and purified by column chromatography [eluent: petroleum ether ~ petroleum ether/ethyl acetate (95/5)] to obtain a colorless oily product (R)-(1-(3-cyclopropylbenzene) (Yl)ethyl) tert-butyl carbamate (430 mg, yield 69%).
第三步：(R)-1-(3-环丙基苯基)乙烷-1-胺盐酸盐的制备The third step: Preparation of (R)-1-(3-cyclopropylphenyl)ethane-1-amine hydrochloride
Figure PCTCN2020082042-appb-000133
Figure PCTCN2020082042-appb-000133
(R)-(1-(3-环丙基苯基)乙基)氨基甲酸叔丁酯(430mg，1.64mmol)在盐酸乙酸乙酯(10mL，4mol/L)中搅拌16小时。浓缩反应液得到白色固体产物(R)-1-(3-环丙基苯基)乙烷-1-胺盐酸盐(300mg，产率92％)。(R)-(1-(3-Cyclopropylphenyl)ethyl) tert-butyl carbamate (430 mg, 1.64 mmol) was stirred in ethyl acetate hydrochloride (10 mL, 4 mol/L) for 16 hours. The reaction solution was concentrated to obtain a white solid product (R)-1-(3-cyclopropylphenyl)ethane-1-amine hydrochloride (300 mg, yield 92%).
MS m/z(ESI):162.2[M+H] +. MS m/z(ESI): 162.2[M+H] + .
第四步：(R)-N-(1-(3-环丙基苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The fourth step: (R)-N-(1-(3-cyclopropylphenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydro Preparation of imidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000134
Figure PCTCN2020082042-appb-000134
3-(吡啶-4-基)-6-(三氯甲基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑(100mg，0.17mmol)，(R)-1-(3-环丙基苯基)乙烷-1-胺盐酸盐(37mg，0.18mmol)和碳酸钠(212mg，2.52mmol)在乙腈(10mL)和水(5mL)中60℃搅拌1 小时。往反应液里加四氢呋喃(10mL)，再60℃搅拌2小时。反应完，加水(30mL)，用乙酸乙酯(30mL*2)萃取，浓缩有机相后柱层析纯化[洗脱剂:石油醚～石油醚/乙酸乙酯(30:70)]得到粗品(75mg)，再薄层层析纯化(展开剂：石油醚/乙酸乙酯＝1/1)得到黄色固体产物(R)-N-(1-(3-环丙基苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(40mg，产率36％)。3-(pyridin-4-yl)-6-(trichloromethyl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole (100mg, 0.17mmol) , (R)-1-(3-cyclopropylphenyl)ethane-1-amine hydrochloride (37mg, 0.18mmol) and sodium carbonate (212mg, 2.52mmol) in acetonitrile (10mL) and water (5mL) Stir at 60°C for 1 hour. Tetrahydrofuran (10 mL) was added to the reaction solution, followed by stirring at 60°C for 2 hours. After the reaction, add water (30mL), extract with ethyl acetate (30mL*2), concentrate the organic phase, and purify by column chromatography [eluent: petroleum ether ~ petroleum ether/ethyl acetate (30:70)] to obtain the crude product ( 75 mg), and then purified by thin layer chromatography (developing solvent: petroleum ether/ethyl acetate = 1/1) to obtain a yellow solid product (R)-N-(1-(3-cyclopropylphenyl)ethyl)- 3-(Pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (40 mg, yield 36%).
MS m/z(ESI):665.3[M+H] +. MS m/z(ESI): 665.3[M+H] + .
第五步：(R)-N-(1-(3-环丙基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The fifth step: (R)-N-(1-(3-cyclopropylphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f ]Indazole-6-Carboxamide Preparation
Figure PCTCN2020082042-appb-000135
Figure PCTCN2020082042-appb-000135
往(R)-N-(1-(3-环丙基苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(80mg，0.12mmol)的二氯甲烷(2mL)溶液里加三乙基硅烷(28mg，0.24mmol)，三氟乙酸(4mL),室温搅拌1小时。旋干反应液，加甲醇溶解，用氨水调至碱性，浓缩后薄层层析纯化(展开剂：CH 2Cl 2/MeOH＝10/1)得到黄色固体产物(R)-N-(1-(3-环丙基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(26mg，产率51％)。 To (R)-N-(1-(3-cyclopropylphenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ 4,5-f]Indazole-6-carboxamide (80mg, 0.12mmol) in dichloromethane (2mL), add triethylsilane (28mg, 0.24mmol), trifluoroacetic acid (4mL), and stir at room temperature for 1 hour . Rotate the reaction solution to dryness, add methanol to dissolve it, make it alkaline with ammonia water, concentrate and purify by thin layer chromatography (developing solvent: CH 2 Cl 2 /MeOH = 10/1) to obtain a yellow solid product (R)-N-(1 -(3-Cyclopropylphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (26mg, yield Rate 51%).
1H NMR(400MHz,DMSO-d 6)δ13.46-13.39(m,1H),13.12(s,1H),9.40-9.31(m,1H),8.72(s,2H),8.45(s,0.6H),8.12-7.87(m,3H),7.58(s,0.6H),7.23-7.18(m,3H),6.93(d,J＝8Hz,1H),5.20-5.16(m,1H),1.90-1.89(m,1H),1.54(d,J＝8Hz,3H),0.94-0.93(m,2H),0.67-0.66(m,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.46-13.39 (m, 1H), 13.12 (s, 1H), 9.40-9.31 (m, 1H), 8.72 (s, 2H), 8.45 (s, 0.6 H), 8.12-7.87 (m, 3H), 7.58 (s, 0.6H), 7.23-7.18 (m, 3H), 6.93 (d, J = 8Hz, 1H), 5.20-5.16 (m, 1H), 1.90 -1.89 (m, 1H), 1.54 (d, J = 8Hz, 3H), 0.94-0.93 (m, 2H), 0.67-0.66 (m, 2H).
MS m/z(ESI):423.1[M+H]+.MS m/z(ESI): 423.1[M+H]+.
实施例38Example 38
(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(2-((2,2-Difluoroethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000136
Figure PCTCN2020082042-appb-000136
第一步：(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(2-((2,2-difluoroethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl)-1-triphenyl Preparation of methyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000137
Figure PCTCN2020082042-appb-000137
(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(200mg，0.31mmol)、2,2-二氟乙胺(127mg，1.56mmol)于二氯甲烷/甲醇(10mL/5mL)中搅拌，加醋酸3mL，搅拌30min。加氰基硼氢化钠(98mg，1.56mmol)，搅拌过夜，加水，二氯甲烷提取，干燥浓缩至干，柱分离纯化得粗品(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(38mg)。(S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5- f] Indazole-6-carboxamide (200mg, 0.31mmol), 2,2-difluoroethylamine (127mg, 1.56mmol) were stirred in dichloromethane/methanol (10mL/5mL), 3mL of acetic acid was added, and stirred for 30min . Add sodium cyanoborohydride (98mg, 1.56mmol), stir overnight, add water, extract with dichloromethane, dry and concentrate to dryness, column separation and purification to obtain crude product (S)-N-(2-((2,2-difluoro Ethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole- 6-formamide (38 mg).
MS m/z(ESI):704.2[M+H] +. MS m/z(ESI): 704.2[M+H] + .
第二步：(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (S)-N-(2-((2,2-difluoroethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-di Preparation of Hydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000138
Figure PCTCN2020082042-appb-000138
(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(38mg)溶于二氯甲烷(2mL)，加三氟乙酸(2mL)，室温搅拌2h。浓缩至干，柱分离纯化得类白色固体(S)-N-(2-((2,2-二氟乙基)氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(9mg，两步产率6％)。(S)-N-(2-((2,2-Difluoroethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1 ,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide (38mg) was dissolved in dichloromethane (2mL), trifluoroacetic acid (2mL) was added, and the mixture was stirred at room temperature for 2h. Concentrate to dryness, column separation and purification to obtain off-white solid (S)-N-(2-((2,2-difluoroethyl)amino)-1-phenylethyl)-3-(pyridin-4-yl) )-1,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide (9 mg, 6% yield in two steps).
1H NMR(400MHz,DMSO-d 6)δ13.40(s,1H),13.10(s,1H),9.18(s,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.40–7.23(m,5H),7.03(d,J＝10.1Hz,1H),5.71(t,J＝7.0Hz,1H),5.12(dtt,J＝10.4,7.1,1.0Hz,1H),3.28(dt,J＝12.5,7.3Hz,1H),3.17(dt,J＝12.2,7.2Hz,1H),3.12–2.81(m,2H),1.51(p,J＝7.2Hz,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.40 (s, 1H), 13.10 (s, 1H), 9.18 (s, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99(s,1H),7.96–7.89(m,2H),7.40–7.23(m,5H),7.03(d,J=10.1Hz,1H), 5.71(t,J=7.0Hz,1H),5.12 (dtt,J＝10.4,7.1,1.0Hz,1H), 3.28(dt,J＝12.5,7.3Hz,1H), 3.17(dt,J＝12.2,7.2Hz,1H), 3.12–2.81(m,2H ), 1.51(p,J=7.2Hz,1H).
MS m/z(ESI):462.2[M+H] +. MS m/z(ESI): 462.2[M+H] + .
实施例39Example 39
(S)-N-(2-(二甲氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备(S)-N-(2-(Dimethylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole Preparation of -6-formamide
Figure PCTCN2020082042-appb-000139
Figure PCTCN2020082042-appb-000139
第一步：(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000140
Figure PCTCN2020082042-appb-000140
3-(吡啶-4-基)-1-三苯甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸甲酯(2.0g，3.73mmol)、(S)-2-氨基-2-苯基乙烷-1-醇(2.56g，18.66mmol)140℃微波搅拌1h，加水，乙酸乙酯提取，干燥浓缩至干，柱分离纯化得(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(1.46g，产率61％)。3-(pyridin-4-yl)-1-trityl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylic acid methyl ester (2.0g, 3.73mmol), ( S)-2-Amino-2-phenylethane-1-ol (2.56g, 18.66mmol) microwave stirring at 140°C for 1h, add water, extract with ethyl acetate, dry and concentrate to dryness, column separation and purification to obtain (S)- N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole -6-Carboxamide (1.46 g, 61% yield).
MS m/z(ESI):641.2[M+H] +. MS m/z(ESI):641.2[M+H] + .
第二步：(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The second step: (S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ Preparation of 4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000141
Figure PCTCN2020082042-appb-000141
将(S)-N-(2-羟基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(1.46g，2.28mmol)溶于DMSO(10mL)，加IBX(5.1g，18.21mmol)，氮气保护下室温搅拌3h。加二氯甲烷，碳酸钠溶液洗涤，干燥浓缩至干，柱分离纯化得2.52g油状物粗品，直接用于下一步反应。The (S)-N-(2-hydroxy-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5 -f] Indazole-6-carboxamide (1.46g, 2.28mmol) was dissolved in DMSO (10mL), added IBX (5.1g, 18.21mmol), and stirred at room temperature for 3h under nitrogen protection. Add dichloromethane, wash with sodium carbonate solution, dry and concentrate to dryness, column separation and purification to obtain 2.52g crude oil, which is directly used in the next reaction.
MS m/z(ESI):639.2[M+H] +. MS m/z(ESI): 639.2[M+H] + .
第三步：(S)-N-(2-(二甲氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The third step: (S)-N-(2-(dimethylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-di Preparation of Hydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000142
Figure PCTCN2020082042-appb-000142
(S)-N-(2-羰基-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(200mg，0.31mmol)，2M的二甲胺甲醇溶液(1.57mL，3.13mmol)于二氯甲烷/甲醇(10mL/5mL)中搅拌，加醋酸0.5mL，搅拌30min。加氰基硼氢化钠(98mg，1.56mmol)，搅拌过夜，加水，二氯甲烷提取，干燥浓缩至干，柱分离纯化得油状物(66mg)，直接用于下步反应。(S)-N-(2-carbonyl-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5- f] Indazole-6-carboxamide (200mg, 0.31mmol), 2M dimethylamine methanol solution (1.57mL, 3.13mmol) was stirred in dichloromethane/methanol (10mL/5mL), 0.5mL of acetic acid was added, and stirred 30min. Add sodium cyanoborohydride (98mg, 1.56mmol), stir overnight, add water, extract with dichloromethane, dry and concentrate to dryness, column separation and purification to obtain an oil (66mg), which is directly used in the next step.
MS m/z(ESI):668.3[M+H] +. MS m/z(ESI): 668.3[M+H] + .
第四步：(S)-N-(2-(二甲氨基)-1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The fourth step: (S)-N-(2-(dimethylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5- f] Preparation of indazole-6-carboxamide
将(S)-N-(2-(二甲氨基)-1-苯基乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(66mg)溶于二氯甲烷(5mL)，加三氟乙酸(2mL)，室温搅拌1h。浓缩至干，柱分离纯化得类白色固体(10mg，两步收率8％)。(S)-N-(2-(Dimethylamino)-1-phenylethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo [4,5-f]Indazole-6-carboxamide (66 mg) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added, and the mixture was stirred at room temperature for 1 h. Concentrate to dryness, column separation and purification to obtain an off-white solid (10 mg, two-step yield 8%).
1H NMR(400MHz,DMSO-d 6)δ13.40(s,1H),13.10(s,1H),9.18(s,1H),8.72(d,J＝5.7Hz,2H),8.50(s,0.5H),8.29–7.81(m,3H),7.58(s,0.5H),7.48(d,J＝7.4Hz,2H),7.34(t,J＝7.5Hz,2H),7.26(t,J＝7.3Hz,1H),5.17(s,1H),2.96(s,1H),2.43(dd,J＝12.6,5.2Hz,1H),2.24(s,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.40 (s, 1H), 13.10 (s, 1H), 9.18 (s, 1H), 8.72 (d, J = 5.7 Hz, 2H), 8.50 (s, 0.5H),8.29–7.81(m,3H),7.58(s,0.5H),7.48(d,J=7.4Hz,2H),7.34(t,J=7.5Hz,2H),7.26(t,J =7.3Hz,1H), 5.17(s,1H), 2.96(s,1H), 2.43(dd,J=12.6,5.2Hz,1H), 2.24(s,6H).
MS m/z(ESI):426.2[M+H] +. MS m/z(ESI): 426.2[M+H] + .
实施例40Example 40
(R)-N-(1-(3-异丙基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(R)-N-(1-(3-isopropylphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole- 6-formamide
Figure PCTCN2020082042-appb-000143
Figure PCTCN2020082042-appb-000143
第一步：(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The first step: (R)-N-(1-(3-bromophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo Preparation of [4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000144
Figure PCTCN2020082042-appb-000144
3-(吡啶-4-基)-6-(三氯甲基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑(2g，3.35mmol)，(R)-1-(3-溴苯基)乙胺(737mg，3.69mmol)和碳酸氢钠(4.22g， 50.25mmol)在乙腈(50mL)和水(25mL)中60℃搅拌1小时。往反应液里加四氢呋喃(50mL)，再60℃搅拌2小时。反应完，加水(60mL)，用乙酸乙酯(50mL*2)萃取，浓缩有机相后柱层析[洗脱剂:石油醚～石油醚/乙酸乙酯(30:70)]得到棕色固体产物(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(700mg,产率30％)。3-(pyridin-4-yl)-6-(trichloromethyl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole (2g, 3.35mmol) , (R)-1-(3-bromophenyl)ethylamine (737 mg, 3.69 mmol) and sodium bicarbonate (4.22 g, 50.25 mmol) were stirred in acetonitrile (50 mL) and water (25 mL) at 60°C for 1 hour. Tetrahydrofuran (50 mL) was added to the reaction solution, followed by stirring at 60°C for 2 hours. After the reaction, add water (60mL), extract with ethyl acetate (50mL*2), concentrate the organic phase and column chromatography [eluent: petroleum ether ~ petroleum ether/ethyl acetate (30:70)] to obtain a brown solid product (R)-N-(1-(3-Bromophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5 -f] Indazole-6-carboxamide (700 mg, 30% yield).
MS m/z(ESI):703.1[M+H] +. MS m/z(ESI): 703.1[M+H] + .
第二步：(R)-N-(1-(3-(丙-1-烯-2-基)苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Step 2: (R)-N-(1-(3-(Pro-1-en-2-yl)phenyl)ethyl)-3-(pyridin-4-yl)-1-trityl Of phenyl-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000145
Figure PCTCN2020082042-appb-000145
(R)-N-(1-(3-溴苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(200mg，0.28mmol)，4,4,5,5-四甲基-2-(丙-1-烯-2-基)-1,3,2-二噁硼戊环(191mg，1.14mmol)，[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(23mg，0.028mmol)和碳酸钾(118mg，0.85mmol)在二氧六环(6mL)和水(0.6mL)中微波130℃反应1.5小时。浓缩反应液后柱层析纯化[洗脱剂:石油醚～石油醚/乙酸乙酯(30:70)]得到棕色油状产物(R)-N-(1-(3-(丙-1-烯-2-基)苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(100mg,53％)。(R)-N-(1-(3-Bromophenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5 -f] indazole-6-carboxamide (200mg, 0.28mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-di Oxaborolane (191mg, 1.14mmol), [1,1'-bis(diphenylphosphine)ferrocene] dichloropalladium dichloromethane complex (23mg, 0.028mmol) and potassium carbonate (118mg, 0.85mmol) was reacted in dioxane (6mL) and water (0.6mL) in microwave at 130°C for 1.5 hours. The reaction solution was concentrated and purified by column chromatography [eluent: petroleum ether ~ petroleum ether/ethyl acetate (30:70)] to obtain a brown oily product (R)-N-(1-(3-(prop-1-ene) -2-yl)phenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide (100 mg, 53%).
MS m/z(ESI):665.2[M+H] +. MS m/z(ESI): 665.2[M+H] + .
第三步：(R)-N-(1-(3-异丙基苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺制备The third step: (R)-N-(1-(3-isopropylphenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydro Preparation of imidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000146
Figure PCTCN2020082042-appb-000146
(R)-N-(1-(3-(丙-1-烯-2-基)苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(100mg，0.15mmol)和Pd(OH) 2/C(100mg)在四氢呋喃(6mL)中，氢气球下室温搅拌16小时。过滤反应液，浓缩滤液得到黄黑色固体产物(R)-N-(1-(3-异丙基苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(100mg，产率100％)。 (R)-N-(1-(3-(Pro-1-en-2-yl)phenyl)ethyl)-3-(pyridin-4-yl)-1-trityl-1, 7-dihydroimidazo[4,5-f]indazole-6-carboxamide (100mg, 0.15mmol) and Pd(OH) 2 /C (100mg) in tetrahydrofuran (6mL), stirring at room temperature under a hydrogen balloon 16 hour. The reaction solution was filtered, and the filtrate was concentrated to obtain a yellow-black solid product (R)-N-(1-(3-isopropylphenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl -1,7-Dihydroimidazo[4,5-f]indazole-6-carboxamide (100 mg, yield 100%).
MS m/z(ESI):667.3[M+H]+.MS m/z(ESI):667.3[M+H]+.
第四步：(R)-N-(1-(3-异丙基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备The fourth step: (R)-N-(1-(3-isopropylphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f ]Indazole-6-Carboxamide Preparation
Figure PCTCN2020082042-appb-000147
Figure PCTCN2020082042-appb-000147
往(R)-N-(1-(3-异丙基苯基)乙基)-3-(吡啶-4-基)-1-三苯代甲基-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(100mg，0.15mmol)的二氯甲烷(2mL)溶液里加三乙基硅烷(35mg，0.30mmol)，三氟乙酸(4mL)，室温搅拌1小时。旋干反应液后，加甲醇溶解，用氨水调至碱性，浓缩后薄层层析(展开剂：CH 2Cl 2/MeOH＝10/1)得到黄色固体产物(R)-N-(1-(3-异丙基苯基)乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(28mg，产率44％)。 To (R)-N-(1-(3-isopropylphenyl)ethyl)-3-(pyridin-4-yl)-1-tritylmethyl-1,7-dihydroimidazo[ 4,5-f]Indazole-6-carboxamide (100mg, 0.15mmol) in dichloromethane (2mL), add triethylsilane (35mg, 0.30mmol), trifluoroacetic acid (4mL), and stir at room temperature for 1 hour . After spin-drying the reaction solution, add methanol to dissolve it, adjust it to alkaline with ammonia water, concentrate and thin-layer chromatography (developing solvent: CH 2 Cl 2 /MeOH = 10/1) to obtain a yellow solid product (R)-N-(1 -(3-isopropylphenyl)ethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide (28mg, yield Rate 44%).
1H NMR(400MHz,DMSO-d 6)δ13.41(d,J＝28Hz,1H),13.21-13.07(m,1H),9.40-9.29(m,1H),8.73-8.70(m,2H),8.44(s,0.6H),8.12(s,0.4H),8.05-7.97(m,2H),7.87(s,0.4H),7.58(s,0.6H),7.35(s,1H),7.30-7.24(m,2H),7.14-7.12(m,1H),5.23-5.20(m,1H),2.91-2.84(m,1H),1.56(d,J＝8Hz,3H),1.20(d,J＝4Hz,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.41 (d, J = 28Hz, 1H), 13.21-13.07 (m, 1H), 9.40-9.29 (m, 1H), 8.73-8.70 (m, 2H) ,8.44(s,0.6H),8.12(s,0.4H),8.05-7.97(m,2H),7.87(s,0.4H),7.58(s,0.6H),7.35(s,1H),7.30 -7.24(m,2H),7.14-7.12(m,1H),5.23-5.20(m,1H),2.91-2.84(m,1H),1.56(d,J=8Hz,3H), 1.20(d, J=4Hz, 6H).
MS m/z(ESI):425.2[M+H] +. MS m/z(ESI): 425.2[M+H] + .
实施例41Example 41
N-苯甲基-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺的制备Preparation of N-benzyl-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide
Figure PCTCN2020082042-appb-000148
Figure PCTCN2020082042-appb-000148
将甲基3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-羧酸酯(7.0mg，0.0239mmol)、苄胺(1mL)于100℃微波搅拌1h，浓缩至干，柱分离纯化得N-苯甲基-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺(0.8mg，产率9％)。Mix methyl 3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxylate (7.0mg, 0.0239mmol), benzylamine (1mL) in Microwave stirring at 100°C for 1h, concentrated to dryness, column separation and purification to obtain N-benzyl-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6- Formamide (0.8 mg, 9% yield).
1H NMR(400MHz,DMSO-d 6)δ13.40-13.35(m,1H),13.20-13.08(m,1H),8.71–8.64(m,2H),8.13(s,1H),7.99(s,1H),7.96–7.89(m,2H),7.51(s,1H),7.37–7.23(m,5H),4.64(dd,J＝10.0,1.0Hz,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ 13.40-13.35 (m, 1H), 13.20-13.08 (m, 1H), 8.71-8.64 (m, 2H), 8.13 (s, 1H), 7.99 (s ,1H), 7.96–7.89(m,2H), 7.51(s,1H), 7.37–7.23(m,5H), 4.64(dd,J=10.0,1.0Hz,2H).
MS m/z(ESI):369.1[M+H] +. MS m/z(ESI): 369.1[M+H] + .
化合物生物学测试评价Compound biology test evaluation
以下结合测试例进一步描述解释本发明，但这些实施例不限制本发明的保护范围。The following further describes and explains the present invention in conjunction with test examples, but these examples do not limit the protection scope of the present invention.
一、测试酶学实验1. Test the enzyme experiment
1.1本发明化合物对ERK-1激酶活性抑制作用的测定1.1 Determination of the inhibitory effect of the compound of the present invention on ERK-1 kinase activity
该测试例的目的是测量化合物对ERK-1激酶活性的抑制能力。使用LANCE Ultra(Perkin Elmer)方法进行体外ERK-1激酶分析。通过在384孔板(Perkin Elmer OPTIPLATE TM)中添加2.5μL的测试化合物/DMSO(最终4％，V/V，使用1:3稀释方案稀释至10个浓度(400nM至0.02nM))，加入用激酶缓冲液(50mM Hepes pH 7.4，10mM MgC12，1mM EGTA，0.01％Triton X-100，2mM DTT)制备的ERK1酶(Invitrogen，#PV3311，最终浓度：0.4nM)和底物Ulight-MBP peptide(Perkin Elmer，#TRF0109-M，最终浓度：0.5μM)混合液5μL，再加入用上述相同缓冲液制备的2.5μL的ATP(Invitrogen，#PV3227，最终浓度38.15μM)溶液混匀后开始反应(反应体系10μL)。在室温下孵育该反应混合物60分钟。通过加10μL/孔用超纯水稀释的终止缓冲液(1X LANCE Detection buffer(PerkinElmer#CR97-100)，10mM EDTA(Invitrogen#15575038)，1nM Eu-anti-p-MBP(PerkinElmer#TRF0201-M)抗体终止反应。在室温下孵育该板60分钟，在Synergy H1 Hybird Reader，H1MFD(Biotek)酶标仪上用激发波长320nm进行激发读数。通过用受体发射信号(在665nM信号值)除以Eu供体发射信号(在615nM信号值)计算TR-FRET比率。相对于板上Max signal(DMSO对照物)和Min signal(不添加酶)对照物孔计算使用化合物处理的孔的百分比抑制TR-FRET比率数据{％抑制率＝100-[(测试化合物-Min平均值)]/(Max平均值-Min平均值)×100}。化合物的浓度经过反应体系稀释4倍后的10个浓度为100nM至0.005nM。使用GraphPad prism拟合百分比抑制率和十点浓度数据至4参数非线性逻辑公式计算出Abs_IC50值。 The purpose of this test case is to measure the inhibitory ability of the compound on ERK-1 kinase activity. In vitro ERK-1 kinase analysis was performed using the LANCE Ultra (Perkin Elmer) method. By adding 2.5μL of test compound / DMSO in a 384-well plates (Perkin Elmer OPTIPLATE TM) (final 4%, V / V, using a 1: 10 dilution to a concentration (400 nM to 0.02nM) 3 dilution scheme), was added with ERK1 enzyme (Invitrogen, #PV3311, final concentration: 0.4nM) prepared with kinase buffer (50mM Hepes pH 7.4, 10mM MgC12, 1mM EGTA, 0.01% Triton X-100, 2mM DTT) and substrate Ulight-MBP peptide (Perkin) Elmer, #TRF0109-M, final concentration: 0.5μM) mix 5μL, then add 2.5μL of ATP (Invitrogen, #PV3227, final concentration 38.15μM) solution prepared with the same buffer as above, mix well and start the reaction (reaction system) 10μL). The reaction mixture was incubated for 60 minutes at room temperature. Stop buffer (1X LANCE Detection buffer (PerkinElmer#CR97-100), 10mM EDTA (Invitrogen#15575038), 1nM Eu-anti-p-MBP (PerkinElmer#TRF0201-M) diluted with ultrapure water by adding 10μL/well The antibody terminates the reaction. Incubate the plate at room temperature for 60 minutes, and perform excitation readings on the Synergy H1 Hybird Reader, H1MFD (Biotek) microplate reader with an excitation wavelength of 320nm. By dividing the receptor emission signal (signal value at 665nM) by Eu Calculate the TR-FRET ratio from the donor emission signal (signal value at 615nM). Calculate the percentage of the wells treated with the compound relative to the Max signal (DMSO control) and Min signal (no enzyme added) control wells on the plate to inhibit TR-FRET Ratio data {% inhibition rate=100-[(test compound-Min average value)]/(Max average value-Min average value)×100}. The concentration of the compound is 4 times diluted by the reaction system at 10 concentrations from 100 nM to 0.005nM. Use GraphPad prism to fit percent inhibition rate and ten-point concentration data to a 4-parameter non-linear logic formula to calculate the Abs_IC50 value.
1.2本发明化合物对ERK2激酶活性抑制作用的测定1.2 Determination of the inhibitory effect of the compound of the present invention on ERK2 kinase activity
该测试例的目的是测量化合物对ERK-2激酶活性的抑制能力。使用LANCE Ultra(Perkin Elmer)方法进行体外ERK-2激酶分析。通过在384孔板(Perkin Elmer OPTIPLATE TM)中添加2.5μL的测试化合物/DMSO(最终4％,V/V，使用1:3稀释方案稀释至10个浓度(400nM至0.02nM))，加入用激酶缓冲液(50mM Hepes pH 7.4，10mM MgC1 2，1mM EGTA，0.01％Triton X-100，2mM DTT)制备的ERK-2酶(Invitrogen，#PV3313，最终浓度：0.08nM)和底物Ulight-MBP peptide(Perkin Elmer,#TRF0109-M，最终浓度：0.5μM)混合液5μL，再加入用上述相同缓冲液制备的2.5μL的ATP(Invitrogen，#PV3227，最终浓度38.15μM)溶液混匀后开始反应(反应体系10μL)。在室温下孵育该反应混合物60分钟。通过加10μL/孔用超纯水稀释的终止缓冲液(1X LANCE Detection buffer(PerkinElmer#CR97-100)，10mM EDTA(Invitrogen#15575038)，1nM Eu-anti-p-MBP (PerkinElmer#TRF0201-M)抗体终止反应。在室温下孵育该板60分钟。在Synergy H1 Hybird Reader，H1MFD(Biotek)酶标仪上用激发波长320nm进行激发读数。 通过用受体发射信号(在665nM信号值)除以Eu供体发射信号(在615nM信号值)计算TR-FRET比率。相对于板上Max signal(DMSO对照物)和Min signal(不添加酶)对照物孔计算使用化合物处理的孔的百分比抑制TR-FRET比率数据{％抑制率＝100-[(测试化合物-Min平均值)]/(Max平均值-Min平均值)×100}。化合物的浓度经过反应体系稀释4倍后的10个浓度为100nM至0.005nM。使用GraphPad prism拟合百分比抑制率和十点浓度数据至4参数非线性逻辑公式计算出Abs_IC 50值。 The purpose of this test case is to measure the compound's ability to inhibit ERK-2 kinase activity. In vitro ERK-2 kinase analysis was performed using LANCE Ultra (Perkin Elmer) method. By adding 2.5 μL of test compound/DMSO (final 4%, V/V, diluted to 10 concentrations (400nM to 0.02nM) using a 1:3 dilution scheme) in a 384-well plate (Perkin Elmer OPTIPLATE TM ), adding ERK-2 enzyme (Invitrogen, #PV3313, final concentration: 0.08nM) prepared in kinase buffer (50mM Hepes pH 7.4, 10mM MgCl 2 , 1mM EGTA, 0.01% Triton X-100, 2mM DTT) and substrate Ulight-MBP Peptide (Perkin Elmer, #TRF0109-M, final concentration: 0.5μM) mixed solution 5μL, then add 2.5μL of ATP (Invitrogen, #PV3227, final concentration 38.15μM) solution prepared with the same buffer as above, mix well and start the reaction (Reaction system 10μL). The reaction mixture was incubated for 60 minutes at room temperature. Stop buffer (1X LANCE Detection buffer (PerkinElmer#CR97-100), 10mM EDTA (Invitrogen#15575038), 1nM Eu-anti-p-MBP (PerkinElmer#TRF0201-M) diluted with ultrapure water by adding 10μL/well The antibody terminates the reaction. Incubate the plate at room temperature for 60 minutes. Read on the Synergy H1 Hybird Reader, H1MFD (Biotek) plate reader with excitation wavelength 320nm. By dividing the receptor emission signal (signal value at 665nM) by Eu Calculate the TR-FRET ratio from the donor emission signal (signal value at 615nM). Calculate the percentage of the wells treated with the compound relative to the Max signal (DMSO control) and Min signal (no enzyme added) control wells on the plate to inhibit TR-FRET Ratio data {% inhibition rate=100-[(test compound-Min average value)]/(Max average value-Min average value)×100}. The concentration of the compound is 4 times diluted by the reaction system at 10 concentrations from 100 nM to 0.005nM. fitted using GraphPad prism percent inhibition and ten-point concentration data to a 4 parameter nonlinear logic formula 50 values were calculated Abs_IC.
通过以上方案得出：本发明所示的化合物在ERK抑制试验中显示出约0.01nM至100nM(IC 50)的生物活性。 According to the above scheme, the compound shown in the present invention shows a biological activity of about 0.01 nM to 100 nM (IC 50 ) in the ERK inhibition test.
在一些实施方案中，本发明的化合物对于ERK-1和/或ERK-2的IC 50小于约100nM、优选化合物小于约10nM、进一步优选小于约5nM、更优选小于约1nM、本发明所列示的化合物中最优选小于0.1nM。在一些实施方案中，本发明的化合物对于ERK的IC 50小于约100nM、优选小于约10nM、进一步优选小于约5nM、更优选小于约1nM、列示化合物中最优选小于0.1nM。在一些其他实施方案中，本发明列示化合物中优选化合物显示出双结合特异性，并且能够以小于约100nM、小于约10nM、小于约5nM、小于约1nM、小于0.1nM的IC 50值抑制ERK激酶(例如，ERK-1激酶、ERK-2激酶等)以及蛋白激酶(例如，Ras、Raf、Her-2、MEK1等)。 In some embodiments, compounds of the invention and for the IC ERK-1 / ERK-2 is 50 or less than about 10OnM, preferably less than about 10 nM compound, more preferably less than about of 5 nM, more preferably less than about 1 nM, the present invention is presented The most preferred compound is less than 0.1 nM. In some embodiments, compounds of the invention for the ERK IC 50 less than about 10OnM, preferably less than about 10 nM, more preferably less than about of 5 nM, more preferably less than about 1 nM, most preferably less than compounds listed 0.1nM. In some other embodiments, the present invention is preferably a compound listed compounds show dual binding specificity, and can be less than about 10OnM, less than about 10 nM, less than about of 5 nM, less than about 1 nM, less than the IC 50 values of 0.1nM of ERK Kinases (eg, ERK-1 kinase, ERK-2 kinase, etc.) and protein kinases (eg, Ras, Raf, Her-2, MEK1, etc.).
具体地，实施例的测试数据见表13：Specifically, the test data of the embodiment is shown in Table 13:
表13：化合物对ERK-1、ERK-2激酶活性抑制相对IC 50值 Table 13: Relative IC 50 value of compound inhibition of ERK-1 and ERK-2 kinase activity
Figure PCTCN2020082042-appb-000149
Figure PCTCN2020082042-appb-000149
Figure PCTCN2020082042-appb-000150
Figure PCTCN2020082042-appb-000150
1.3本发明化合物对肿瘤细胞增殖抑制活性的测定1.3 Determination of the compound of the present invention on tumor cell proliferation inhibitory activity
该测试例的目的是为了测定本发明中的化合物对肿瘤细胞增殖活性的抑制作用。化合物对肿瘤细胞增殖抑制活性通过CellTiter-Glo的方法进行测定，并得出化合物抑制细胞增殖活性的半数抑制浓度IC 50。在96孔细胞培养板中接种50～100μL的肿瘤细胞悬液，密度为1～5*10 4细胞/ml，将培养板于培养箱培养16～24小时(37℃，5％CO 2)。向培养板细胞中加入梯度稀释的不同浓度的待测化合物溶液，将培养板在培养箱孵育3-7天(37℃，5％CO 2)。每孔加入50～100μL CellTiter-Glo试剂，并振荡10分钟，室温静置10分钟。酶标仪测定化学发光信号值。通过化学发光信号值计算抑制率。根据不同浓度的抑制率通过曲线拟合得出化合物的IC 50。 The purpose of this test example is to determine the inhibitory effect of the compound of the present invention on tumor cell proliferation activity. The compound's anti-tumor cell proliferation inhibitory activity was measured by the CellTiter-Glo method, and the half inhibitory concentration IC 50 of the compound's inhibitory cell proliferation activity was obtained. Inoculate 50-100 μL of tumor cell suspension in a 96-well cell culture plate at a density of 1 to 5*10 4 cells/ml, and culture the plate in an incubator for 16 to 24 hours (37° C., 5% CO 2 ). The test compound solutions of different concentrations in gradient dilutions are added to the cells of the culture plate, and the culture plate is incubated in an incubator for 3-7 days (37° C., 5% CO 2 ). Add 50-100μL CellTiter-Glo reagent to each well, shake for 10 minutes, and stand at room temperature for 10 minutes. The microplate reader measures the chemiluminescence signal value. The inhibition rate was calculated from the value of the chemiluminescence signal. According to the inhibition rate of different concentrations, the IC 50 of the compound was obtained by curve fitting.
本发明中化合物对胰腺癌肿瘤细胞Mia Paca 2增殖活性的试验进行测定，测得的IC 50值见表14。 The compounds of the present invention were tested for the proliferation activity of pancreatic cancer tumor cells Mia Paca 2, and the measured IC 50 values are shown in Table 14.
表14：化合物对胰腺癌肿瘤细胞Mia Paca 2增殖活性抑制相对IC 50值 Table 14: Relative IC 50 value of compound inhibiting proliferation activity of pancreatic cancer tumor cell Mia Paca 2
Figure PCTCN2020082042-appb-000151
Figure PCTCN2020082042-appb-000151
结论：本发明化合物对肿瘤细胞增殖活性具有明显的抑制作用。Conclusion: The compound of the present invention has obvious inhibitory effect on tumor cell proliferation activity.
二、小鼠药代动力学测定2. Determination of pharmacokinetics in mice
2.1实验目的：2.1 Experimental purpose:
以Balb/c Mouse(雄性，购自上海杰思捷实验动物有限公司，动物生产许可证号(SCXK(沪)2013-0006N0.311620400001794))为受试动物，研究化合物Balb/c Mouse (male, purchased from Shanghai Jiesjie Laboratory Animal Co., Ltd., animal production license number (SCXK (Shanghai) 2013-0006N0.311620400001794)) was used as the test animal to study the compound
实施例1、实施例2、实施例6在Mouse体内(血浆)的药代动力学行为。The pharmacokinetic behavior of Example 1, Example 2, and Example 6 in Mouse (plasma).
2.2实验方案：2.2 Experimental program:
称取170g PEG400和20g Solutol HS-15加入250mL玻璃瓶，再加入10mL of NMP。超声10分钟，混合均匀成澄清溶液。Weigh 170g PEG400 and 20g Solutol HS-15 into a 250mL glass bottle, and then add 10mL of NMP. Ultrasound for 10 minutes and mix well to form a clear solution.
称取11.2mg实施例1，10.8mg实施例2，9.8mg实施例6，分别加入4mL玻璃瓶，加入1.940mL，2.160mL和1.525mL该溶液，超声10分钟，得到无色澄清溶液，浓度为5mg/mL。禁食一夜后分别PO，剂量为50mg/kg，给药体积10mL/kg。Weigh 11.2mg of Example 1, 10.8mg of Example 2, and 9.8mg of Example 6, respectively into 4mL glass bottles, add 1.940mL, 2.160mL and 1.525mL of this solution, sonicate for 10 minutes to obtain a colorless clear solution with a concentration of 5mg/mL. After fasting overnight, PO respectively, the dose was 50 mg/kg, and the administration volume was 10 mL/kg.
2.3实验结果：2.3 Experimental results:
自由碱化合物的PK实验结果如下表15所示：The PK experiment results of free base compounds are shown in Table 15 below:
表15：自由碱化合物的小鼠药代动力学参数Table 15: Mouse pharmacokinetic parameters of free base compounds
Figure PCTCN2020082042-appb-000152
Figure PCTCN2020082042-appb-000152
由表15可知，50mg/kg剂量下，实施例1，实施例2和实施例6在小鼠血浆中的暴露量良好。It can be seen from Table 15 that at a dose of 50 mg/kg, the exposure of Example 1, Example 2 and Example 6 in mouse plasma is good.
三、MiaPaca 2移植瘤模型上对肿瘤的抑制实验3. Tumor suppression experiment on MiaPaca 2 transplanted tumor model
3.1实验目的：3.1 The purpose of the experiment:
以BALB/c裸小鼠为受试动物，采用人胰腺癌细胞MiaPaca 2异种移植瘤(CDX)模型进行体内药效实验，评价受试化合物抗肿瘤作用。BALB/c nude mice were used as test animals, and the human pancreatic cancer cell MiaPaca 2 xenograft (CDX) model was used for in vivo pharmacodynamic experiments to evaluate the anti-tumor effects of the test compounds.
3.2实验仪器与试剂：3.2 Experimental equipment and reagents:
3.2.1仪器：3.2.1 Instrument:
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(Thermo-311，Thermo) CO 2 incubator (Thermo-311, Thermo)
离心机(Centrifuge 5720R，Eppendorf)Centrifuge (Centrifuge 5720R, Eppendorf)
全自动细胞计数仪(Countess II，Life Technologies)Fully automatic cell counter (Countess II, Life Technologies)
移液器(10-20μL，Eppendorf)Pipette (10-20μL, Eppendorf)
显微镜(Ts 2，尼康)Microscope (Ts 2, Nikon)
游标卡尺(CD-6”AX，日本三丰)Vernier caliper (CD-6"AX, Mitutoyo, Japan)
细胞培养瓶(T25/T75/T225，Corning)Cell culture flask (T25/T75/T225, Corning)
恒温水槽(HWS12，上海一恒科学)Constant temperature water tank (HWS12, Shanghai Yiheng Science)
3.2.2试剂：3.2.2 Reagents:
DMEM(11995-065，Gibco)DMEM (11995-065, Gibco)
胎牛血清(FBS)(10091-148，Gibco)Fetal Bovine Serum (FBS) (10091-148, Gibco)
0.25％胰蛋白酶(25200-056，Gibco)0.25% Trypsin (25200-056, Gibco)
青链霉素双抗(P/S)(SV30010，GE)Penicillin double antibody (P/S) (SV30010, GE)
磷酸盐缓冲液(PBS)(10010-023，Gibco)Phosphate buffered saline (PBS) (10010-023, Gibco)
Matrigel(356234，Corning)Matrigel (356234, Corning)
Gln(25030-081，Gibco)Gln(25030-081, Gibco)
3.3实验操作：3.3 Experimental operation:
从细胞库中取出MiaPaca 2细胞，复苏后加入DMEM培养基(含10％FBS、1％Glu、1％P/S)置于CO 2培养箱中培养(培养箱温度为37℃，CO 2浓度为5％)。待细胞铺满培养瓶底部80-90％后传代，传代后细胞继续置于CO 2培养箱中培养。重复该过程直到细胞数满足体内药效接种需求量，开始收集对数生长期的细胞，用全自动细胞计数仪计数，根据计数结果用PBS和Matrigel(体积比为1：1)重悬细胞，制成细胞悬液(密度8悬细胞 7/ml)，置于冰盒中待用。 Take out MiaPaca 2 cells from the cell bank, add DMEM medium (containing 10% FBS, 1% Glu, 1% P/S) after resuscitation and place them in a CO 2 incubator (the temperature of the incubator is 37℃, and the CO 2 concentration Is 5%). After the cells are 80-90% of the bottom of the culture flask, the cells are passaged. After passage, the cells are continued to be cultured in a CO 2 incubator. Repeat the process until the number of cells meets the requirement of inoculation in vivo, start to collect the cells in the logarithmic growth phase, count them with an automatic cell counter, and resuspend the cells with PBS and Matrigel (volume ratio 1:1) according to the counting results. Prepare cell suspension (density 8 suspended cells 7 /ml) and place in an ice box for later use.
使用动物为BALB/c裸小鼠，雌性，6-8周龄，体重约为18-22克。将小鼠保持在一个无特殊病原体的环境中，且在单个通风笼中，每笼5只小鼠。所有的笼子、垫料和水在使用前进行消毒，所有动物可以自由获取标准认证的商业实验室饮食。实验开始前用一次性大小鼠通用耳标标记裸鼠，接种前用75％医用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8*10 6个细胞)MiaPaca 2肿瘤细胞。当平均肿瘤体积达到100-200mm 3时开始分组给药。受试化合物每日经口灌胃给药，给药剂量、给药频次及实验结束时各组药效情况见表16和表17。每周两次用游标卡尺测量肿瘤体积(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。抗肿瘤药效是通过化合物处理过的动物的平均肿瘤增加体积除以未处理过动物的平均肿瘤增加体积来确定。抑瘤率计算公式为：TGI(％)＝1-[(Vt-V0)给药组/(Vt-V0)溶剂对照组]*100％。实验结束后将所有动物安乐死。 The animals used are BALB/c nude mice, female, 6-8 weeks old, weighing about 18-22 grams. The mice were kept in an environment free of special pathogens and in a single ventilated cage with 5 mice per cage. All cages, litter and water are disinfected before use, and all animals can freely access standard certified commercial laboratory food. Before the experiment, the nude mice were labeled with disposable universal ear tags for rats and mice, and the skin of the inoculation site was disinfected with 75% medical alcohol before inoculation. Each mouse was inoculated with 0.1ml (containing 8*10 6 cells) MiaPaca 2 subcutaneously on the right back of each mouse. Tumor cells. When the average tumor volume reaches 100-200mm 3 , group administration is started. The test compound was administered by oral gavage every day, and the dosage, frequency of administration and the efficacy of each group at the end of the experiment are shown in Table 16 and Table 17. The tumor volume (mm 3 ) was measured with a vernier caliper twice a week, and the calculation formula was: V=0.5*D*d*d, where D and d are the long diameter and short diameter of the tumor, respectively. The antitumor efficacy is determined by dividing the average tumor increase volume of animals treated with the compound by the average tumor increase volume of untreated animals. The calculation formula of the tumor inhibition rate is: TGI(%)=1-[(Vt-V0) administration group/(Vt-V0) solvent control group]*100%. After the experiment, all animals were euthanized.
表16：化合物的移植瘤小鼠药效参数Table 16: Pharmacodynamic parameters of the compounds in transplanted tumor mice
Figure PCTCN2020082042-appb-000153
Figure PCTCN2020082042-appb-000153
Figure PCTCN2020082042-appb-000154
Figure PCTCN2020082042-appb-000154
结果表明，口服连续给药21天后，实施例2在100mg/kg QD和50mg/kg BID两种给药条件下都能显著抑制MiaPaca 2裸小鼠移植瘤生长，药效良好；实施例6在100mg/kg QD给药条件下也能明显抑制肿瘤生长。The results showed that after continuous oral administration for 21 days, Example 2 can significantly inhibit the growth of transplanted tumors in nude mice of MiaPaca 2 under the conditions of 100mg/kg QD and 50mg/kg BID, and the drug effect is good; Example 6 100mg/kg QD can also significantly inhibit tumor growth.
表17：化合物的移植瘤小鼠药效参数Table 17: Pharmacodynamic parameters of the compounds in transplanted tumor mice
Figure PCTCN2020082042-appb-000155
Figure PCTCN2020082042-appb-000155
结果表明，口服连续给药16天后，实施例1在50mg/kg QD给药条件下能显著抑制MiaPaca 2裸小鼠移植瘤生长，药效良好。The results showed that after 16 days of continuous oral administration, Example 1 can significantly inhibit the growth of transplanted tumors in MiaPaca 2 nude mice under the conditions of 50 mg/kg QD administration, and the drug effect is good.
四、hERG钾离子通道抑制活性测试Four, hERG potassium channel inhibitory activity test
4.1细胞准备4.1 Cell preparation
4.1.1 CHO-hERG细胞培养于175cm2培养瓶中，待细胞密度生长到60～80％，移走培养液，用7mL PBS洗一遍，然后加入3mL Detachin消化。4.1.1 CHO-hERG cells are cultured in a 175cm2 culture flask. After the cell density grows to 60-80%, remove the culture solution, wash it with 7mL PBS, and then add 3mL Detachin for digestion.
4.1.2待消化完全后加入7mL培养液中和，然后离心，吸走上清液，再加入5mL培养液重悬，以确保细胞密度为2～5重悬，以确保细。4.1.2 After the digestion is complete, add 7 mL of culture medium to neutralize, then centrifuge, aspirate the supernatant, and then add 5 mL of culture medium to resuspend to ensure that the cell density is 2 to 5 and resuspend to ensure fineness.
4.2溶液配制4.2 Solution preparation
表18：细胞内液和外液的组成成分Table 18: Composition of intracellular fluid and external fluid
Figure PCTCN2020082042-appb-000156
Figure PCTCN2020082042-appb-000156
4.3电生理记录过程4.3 Electrophysiological recording process
单细胞高阻抗封接和全细胞模式形成过程全部由Qpatch仪器自动完成，在获得全细胞记录模式后，细胞钳制在-80毫伏，在给予一个5秒的+40毫伏去极化刺激前，先给予一个50毫秒的-50毫伏前置电压，然后复极化到-50毫伏维持5秒，再回到-80毫伏。每15秒施加此电压刺激，记录2分钟后给予细胞外液记 录5分钟，然后开始给药过程，化合物浓度从最低测试浓度开始，每个测试浓度给予2.5分钟。每个浓度至少测试3个细胞。The single-cell high-impedance sealing and the whole-cell mode formation process are all automatically completed by the Qpatch instrument. After the whole-cell recording mode is obtained, the cell is clamped at -80 mV, before giving a 5 second +40 mV depolarization stimulus , First give a pre-voltage of -50 millivolts for 50 milliseconds, then repolarize to -50 millivolts for 5 seconds, and then return to -80 millivolts. This voltage stimulus was applied every 15 seconds, recorded for 2 minutes, and then administered extracellular fluid for 5 minutes, and then the administration process was started. The compound concentration started from the lowest test concentration, and each test concentration was given for 2.5 minutes. At least 3 cells are tested at each concentration.
4.4化合物准备4.4 Compound preparation
4.4.1将20mM的化合物母液用细胞外液进行稀释，取5uL 20mM的化合物母液加入2495uL细胞外液，500倍稀释至40uM，然后在含0.2％DMSO的细胞外液中依次进行3倍连续稀释得到需要测试的最终浓度。4.4.1 Dilute the 20mM compound mother liquor with extracellular fluid, take 5uL 20mM compound mother liquor and add 2495uL extracellular fluid, dilute 500 times to 40uM, and then successively perform 3-fold serial dilutions in 0.2% DMSO extracellular fluid Get the final concentration that needs to be tested.
4.4.2最高测试浓度为40uM，依次分别为40，13.33，4.44，1.48，0.49，0.16uM共6个浓度。4.4.2 The highest test concentration is 40uM, which are respectively 40, 13.33, 4.44, 1.48, 0.49, 0.16uM and 6 concentrations.
4.4.3最终测试浓度中的DMSO含量不超过0.2％，此浓度的DMSO对hERG钾通道没有影响。4.4.3 The DMSO content in the final test concentration does not exceed 0.2%. This concentration of DMSO has no effect on the hERG potassium channel.
4.5数据分析4.5 Data analysis
实验数据由XLFit软件进行分析。The experimental data is analyzed by XLFit software.
4.6质量控制4.6 Quality control
环境：湿度20～50％，温度22～25％。要测Environment: Humidity is 20-50%, temperature is 22-25%. To test
试剂：所用实验试剂购买于Sigma公司，纯度>98％Reagents: The experimental reagents used were purchased from Sigma, with a purity of >98%
报告中的实验数据必须满足以下标准：The experimental data in the report must meet the following criteria:
全细胞封接阻抗>100M必须满Whole cell sealing impedance>100M must be full
尾电流幅度>400pATail current amplitude>400pA
药理学参数：Pharmacological parameters:
多浓度Cisapride对hERG通道的抑制效应设为阳性对照The inhibitory effect of multiple concentrations of Cisapride on hERG channels was set as a positive control
4.7实验结果4.7 Experimental results
实施例在多浓度对hERG电流的抑制结果：Examples of the inhibition results of hERG current at multiple concentrations:
表19：实施例在多浓度对hERG电流的抑制结果Table 19: Inhibition results of hERG currents at multiple concentrations in Examples
实施例编号Example number hERG(uM)hERG(uM)
实施例2Example 2 >40>40
药物对于心脏hERG钾离子通道的抑制是药物导致QT延长综合症的主要原因。从实验结果可以看出，实施例2，对于心脏hERG钾离子通道没有抑制作用，可以避免高剂量时的心脏毒副作用。The inhibitory effect of drugs on the cardiac hERG potassium channel is the main cause of QT prolonged syndrome caused by drugs. It can be seen from the experimental results that Example 2 has no inhibitory effect on the cardiac hERG potassium ion channel, and can avoid cardiac toxic side effects at high doses.
总之，本发明提供了一系列具有新颖结构的高活性、高选择性ERK1/2激酶抑制剂。在大鼠和小鼠模型上都显示出很好的药代动力学性质，在Miapaca荷瘤小鼠模型上也体现出了良好的药效，有很大的潜力被开发成针对肿瘤类疾病的药物。In summary, the present invention provides a series of highly active and selective ERK1/2 kinase inhibitors with novel structures. It has shown good pharmacokinetic properties in both rat and mouse models, and has also shown good pharmacodynamics in the Miapaca tumor-bearing mouse model. It has great potential to be developed for tumor diseases. drug.
化合物自由碱及其盐的晶型研究Study on the crystal form of compound free base and its salt
1.实施例2自由碱不同晶型的制备1. Example 2 Preparation of different crystal forms of free base
1.1实施例2自由碱晶型I的制备1.1 Example 2 Preparation of free base crystal form I
将4.7g粗品(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺和47mL二氯甲烷加入100mL反应瓶中，冰水浴降温。向反应液中滴加2.4mL三乙基硅烷，13.8mL三氟乙酸，反应液升至室温，搅拌反应5小时。取样，检测反应完全，浓缩反应液除去溶剂。柱层析纯化，得到0.43g黄色固体，向固体中加入8.6mL甲醇，加热至回流1小时，降温搅拌3小时，过滤，滤饼使用乙酸乙酯，甲基叔丁基醚洗涤，干燥滤饼，得到0.23g黄色固体(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺，纯度为99.3％。经检测分析，其为自由碱晶型I，具有如图1所示的XRPD图、如图2所示的TGA图及如图3所示的DSC图。4.7g crude product (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-methan The amide and 47 mL of dichloromethane were added to a 100 mL reaction flask, and the temperature was cooled in an ice water bath. 2.4 mL of triethylsilane and 13.8 mL of trifluoroacetic acid were added dropwise to the reaction solution, the reaction solution was raised to room temperature, and the reaction was stirred for 5 hours. Sampling was taken to detect the completion of the reaction, and the reaction solution was concentrated to remove the solvent. Purified by column chromatography to obtain 0.43g of yellow solid, add 8.6mL methanol to the solid, heat to reflux for 1 hour, cool and stir for 3 hours, filter, wash the filter cake with ethyl acetate, methyl tert-butyl ether, and dry the filter cake , To obtain 0.23 g of yellow solid (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6 -Formamide with a purity of 99.3%. After detection and analysis, it is a free base crystal form I with an XRPD pattern as shown in FIG. 1, a TGA pattern as shown in FIG. 2, and a DSC pattern as shown in FIG. 3.
1.2实施例2自由碱晶型II的制备1.2 Example 2 Preparation of free base crystal form II
将18g粗品(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺和180mL二氯甲烷加入250mL反应瓶中，冰水浴降温，滴加9.2mL三乙基硅烷，55.2mL三氟乙酸，撤去冰水浴，反应液升至室温，搅拌17小时。取样，检测反应完全，浓缩反应液，除去溶剂，浓缩剩余物中加入72mL甲醇，然后冰水浴降温反应液，滴加20mL 4M的氯化氢-乙酸乙酯溶液，滴加完毕，再向反应液中滴加100mL乙酸乙酯，室温搅拌2小时，过滤，滤饼用乙酸乙酯洗涤，干燥得到12.1g土黄色固体。固体中加入180mL 2-甲基四氢呋喃和180mL 2N的氢氧化钠水溶液，搅拌分层，水层再用180mL 2-甲基四氢呋喃萃取，合并有机相，干燥，浓缩除去溶剂，得到8.6g黄色固体。再向固体中加入50mL甲醇和20mL四氢呋喃溶清，随后加入10mL 30％的氯化氢-甲醇溶液，然后再滴加50mL MTBE，室温搅拌1小时，过滤，滤饼(湿品)加入80mL 2-甲基四氢呋喃和80mL 2N的氢氧化钠水溶液，搅拌分层，水层再用80mL 2-甲基四氢呋喃萃取，合并有机相，干燥，浓缩除去溶剂，浓缩剩余物中加入40mL MTBE，室温搅拌1小时，过滤，干燥，得到5.0g浅黄色固体(R)-N-(1-苯基乙基)-3-(吡啶-4-基)-1,7-二氢咪唑并[4,5-f]吲唑-6-甲酰胺，纯度为99.4％。经检测分析，其自由碱晶型II，具有如图4所示的XRPD图、如图5所示的TGA图及如图6所示的DSC图。Put 18g crude product (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indazole-6-carboxamide Add 180 mL of dichloromethane to a 250 mL reaction flask, cool in an ice water bath, add 9.2 mL of triethylsilane and 55.2 mL of trifluoroacetic acid dropwise, remove the ice water bath, and warm the reaction solution to room temperature and stir for 17 hours. Take a sample and check that the reaction is complete. Concentrate the reaction solution and remove the solvent. Add 72mL methanol to the concentrated residue, then cool the reaction solution in an ice-water bath, add 20mL 4M hydrogen chloride-ethyl acetate solution dropwise, and then drop it into the reaction solution. Add 100 mL of ethyl acetate, stir at room temperature for 2 hours, filter, wash the filter cake with ethyl acetate, and dry to obtain 12.1 g of a yellowish solid. Add 180 mL of 2-methyltetrahydrofuran and 180 mL of 2N sodium hydroxide aqueous solution to the solid, stir and separate the layers, and then extract the aqueous layer with 180 mL of 2-methyltetrahydrofuran, combine the organic phases, dry, and concentrate to remove the solvent to obtain 8.6 g of yellow solid. Then add 50mL methanol and 20mL tetrahydrofuran to the solid to dissolve it, then add 10mL 30% hydrogen chloride-methanol solution, then add 50mL MTBE dropwise, stir at room temperature for 1 hour, filter, filter cake (wet product) add 80mL 2-methyl Tetrahydrofuran and 80mL 2N sodium hydroxide aqueous solution were stirred and separated. The aqueous layer was extracted with 80mL 2-methyltetrahydrofuran. The organic phases were combined, dried, concentrated to remove the solvent. The concentrated residue was added with 40mL MTBE, stirred at room temperature for 1 hour, and filtered And dried to obtain 5.0 g of light yellow solid (R)-N-(1-phenylethyl)-3-(pyridin-4-yl)-1,7-dihydroimidazo[4,5-f]indole Azole-6-carboxamide with a purity of 99.4%. After detection and analysis, its free base crystal form II has an XRPD pattern as shown in FIG. 4, a TGA pattern as shown in FIG. 5, and a DSC pattern as shown in FIG. 6.
2.化合物盐型晶型筛选2. Screening of compound salt crystal form
2.1实验目的：2.1 Experimental purpose:
选择不同的反离子酸，通过合适的结晶方法，制备具有晶型的不同盐。Choose different counter-ion acids and prepare different salts with crystal forms through appropriate crystallization methods.
2.2实验仪器和设备：2.2 Experimental instruments and equipment:
名称name 型号model 来源source
分析天平Analytical Balances BSA224S-CWBSA224S-CW SartoriusSartorius
超声波清洗仪Ultrasonic cleaner SK5200LHCSK5200LHC 上海科导超声仪器Shanghai Kedao Ultrasound Instrument
移液枪Pipette Eppendorf(50mL,1000μL)Eppendorf (50mL, 1000μL) EppendorfEppendorf
2.3实验步骤：2.3 Experimental steps:
2.3.1实施例2的盐型晶型筛选2.3.1 The salt crystal form screening of Example 2
①马来酸盐晶型I的制备① Preparation of maleate crystal form I
称取500mg实施例2自由碱，加入5mL N,N-二甲基甲酰胺超声未溶清，体系在50℃下加热至完全溶清，另称取183mg马来酸加1mL甲醇室温振荡完全溶清。在50℃下将马来酸的甲醇溶液加入碱的溶液中，很快析出沉淀，搅拌0.5h后向体系中加入10mL甲基叔丁基醚，继续搅拌1.5h后关闭加热，冷却至室温，最后过滤，滤饼用20mL甲基叔丁基醚淋洗，固体放入40℃真空干燥箱烘干至恒重，得到马来酸盐晶型I。经检测分析，其具有如图7所示的XRPD图、如图8所示的TGA图及如图9所示的DSC图。Weigh 500 mg of the free base of Example 2 and add 5 mL of N,N-dimethylformamide ultrasonically undissolved. The system is heated at 50°C until it is completely dissolved. Separately weigh 183 mg of maleic acid and 1 mL of methanol to completely dissolve at room temperature. clear. The methanol solution of maleic acid was added to the alkali solution at 50°C, and the precipitate precipitated out quickly. After stirring for 0.5h, 10mL methyl tert-butyl ether was added to the system. After stirring for 1.5h, the heating was turned off and the temperature was cooled to room temperature. Finally, it is filtered, the filter cake is rinsed with 20 mL of methyl tert-butyl ether, and the solid is placed in a vacuum drying oven at 40° C. and dried to a constant weight to obtain maleate crystal form I. After detection and analysis, it has an XRPD chart as shown in FIG. 7, a TGA chart as shown in FIG. 8 and a DSC chart as shown in FIG. 9.
②马来酸盐晶型II的制备② Preparation of maleate crystal form II
称取498.37mg实施例2自由碱，加入10mL甲醇，50℃搅拌不溶，为灰白色混悬液。另称取185.05mg马来酸加1mL甲醇超声溶清。将马来酸的甲醇溶液加入50℃的自由碱混悬液中，混悬液立即由灰白色变成黄色。在50℃下搅拌反应2h后关闭加热，将至室温，最后过滤，滤饼用10mL甲基叔丁基醚淋洗。固体放入40℃真空干燥箱烘干至恒重，得到马来酸盐晶型II。经检测分析，其具有如图10所示的XRPD图及如图11所示的DSC图。Weigh 498.37 mg of the free base of Example 2, add 10 mL of methanol, stir at 50°C and insoluble, and become an off-white suspension. Separately weigh 185.05 mg maleic acid and add 1 mL methanol to ultrasonically dissolve it. The methanol solution of maleic acid was added to the free base suspension at 50°C, and the suspension immediately changed from off-white to yellow. After stirring the reaction at 50°C for 2 hours, the heating was turned off, the temperature was brought to room temperature, and finally filtered, and the filter cake was rinsed with 10 mL of methyl tert-butyl ether. The solid was dried in a vacuum drying oven at 40°C to a constant weight to obtain maleate crystal form II. After detection and analysis, it has an XRPD chart as shown in FIG. 10 and a DSC chart as shown in FIG. 11.
③马来酸盐晶型III的制备③ Preparation of maleate crystal form III
称取30mg马来酸盐，加入200uL四氢呋喃，在50℃下打浆4d，最后将固体离心，去除上清液后固体放入40℃真空干燥箱中烘干至恒重，得到马来酸盐晶型III。经检测分析，其具有如图12所示的XRPD图及如图13所示的DSC图。Weigh 30mg of maleate, add 200uL of tetrahydrofuran, beat at 50℃ for 4d, finally centrifuge the solid, remove the supernatant, and place the solid in a vacuum drying oven at 40℃ to dry to constant weight to obtain maleate crystals Type III. After detection and analysis, it has an XRPD chart as shown in FIG. 12 and a DSC chart as shown in FIG. 13.
④马来酸盐晶型IV的制备④ Preparation of maleate crystal form IV
称取50.23mg马来酸盐，加入0.75mL N-甲基吡咯烷酮在50℃下完全溶清，向体系中加入1mL异丙醚，很快析出黄色固体，在50℃下反应2h后继续加入2mL异丙醚，并关闭加热将至室温，最后将固体离心，去除上清液后固体放入40℃真空干燥箱中烘干至恒重，得到马来酸盐晶型IV。经检测分析，其具有如图14所示的XRPD图及如图15所示的DSC图。Weigh 50.23mg of maleate, add 0.75mL of N-methylpyrrolidone to completely dissolve it at 50℃, add 1mL of isopropyl ether to the system, and quickly precipitate a yellow solid. After reacting at 50℃ for 2h, continue to add 2mL Isopropyl ether was turned off and the heating was brought to room temperature. Finally, the solid was centrifuged, the supernatant was removed, and the solid was dried in a vacuum drying oven at 40° C. to a constant weight to obtain maleate crystal form IV. After detection and analysis, it has an XRPD chart as shown in FIG. 14 and a DSC chart as shown in FIG. 15.
⑤盐酸盐的制备⑤ Preparation of hydrochloride
称取30mg化合物实施例2自由碱，加入1mL四氢呋喃加热至40℃搅拌不溶，向混悬液中加入94uL 1M盐酸的乙醇溶液，加入后无明显现象，将体系加热至50℃，又加入0.5mL甲醇，在50℃下反应2h后关闭加热，室温搅拌16h后离心，用2mL丙酮振荡离心两次，去除上清液后固体放入40℃真空干燥箱中烘干至恒重，得到盐酸盐晶体。经检测分析，其具有如图16所示的XRPD图。Weigh 30 mg of the free base of compound Example 2, add 1 mL of tetrahydrofuran and heat to 40°C and stir to be insoluble, add 94uL 1M hydrochloric acid in ethanol to the suspension, and there is no obvious phenomenon after adding, heat the system to 50°C, and add another 0.5 mL Methanol, react at 50℃ for 2h, turn off heating, stir at room temperature for 16h, centrifuge, and centrifuge twice with 2mL acetone. After removing the supernatant, the solid is placed in a vacuum drying oven at 40℃ to dry to constant weight to obtain hydrochloride. Crystal. After detection and analysis, it has an XRPD pattern as shown in FIG. 16.
⑥对苯甲磺酸盐晶型I的制备⑥ Preparation of p-toluenesulfonate crystal form I
称取30mg化合物实施例2自由碱，加入1mL甲醇，40℃下搅拌不溶，向混悬液中加入94uL 1.18M对苯甲磺酸的乙醇溶液，超声振荡完全溶清，40℃下搅拌1h仍溶清。将加热关闭，搅拌1h后析出大量黄色固体，室温搅拌48h后离心，用1.5mL甲基叔丁基醚振荡离心两次，去除上清液后固体放入40℃真空干燥箱中烘干至恒重。得到对苯甲磺酸盐晶型I。经检测分析，其具有如图17所示的XRPD图。Weigh 30 mg of the free base of Compound Example 2 and add 1 mL of methanol. Stir insoluble at 40°C. Add 94uL 1.18M ethanolic solution of p-toluenesulfonic acid to the suspension. It will dissolve completely after ultrasonic shaking. After stirring at 40°C for 1h, Dissolve. Turn off the heating, and after stirring for 1 hour, a large amount of yellow solid precipitated. After stirring at room temperature for 48 hours, centrifuge, and centrifuge twice with 1.5 mL methyl tert-butyl ether. After removing the supernatant, the solid was placed in a vacuum drying oven at 40°C and dried until constant weight. The crystal form I of p-toluenesulfonate salt is obtained. After detection and analysis, it has an XRPD pattern as shown in FIG. 17.
⑦对苯甲磺酸盐晶型II的制备⑦ Preparation of p-toluenesulfonate crystal form II
称取30mg对苯甲磺酸盐，加入200uL乙醇，在50℃下打浆4d，最后将固体离心，去除上清液后固体放入40℃真空干燥箱中烘干至恒重，得到对苯甲磺酸盐晶型II。经检测分析，其有如图18所示的XRPD图。Weigh 30mg of p-toluenesulfonate, add 200uL ethanol, and beat at 50°C for 4 days. Finally, centrifuge the solid, remove the supernatant, and place the solid in a vacuum drying oven at 40°C to dry to constant weight to obtain p-benzyl Sulfonate Form II. After detection and analysis, it has an XRPD pattern as shown in Figure 18.
⑧对苯甲磺酸盐晶型III的制备⑧ Preparation of p-toluenesulfonate crystal form III
称取30mg对苯甲磺酸盐，加入200uL88％丙酮，在50℃下打浆4d，最后将固体离心，去除上清液后固体放入40℃真空干燥箱中烘干至恒重，得到对苯甲磺酸盐晶型III。经检测分析，其有如图19所示的XRPD图。Weigh 30 mg of p-toluenesulfonate, add 200uL 88% acetone, and beat at 50°C for 4 days. Finally, centrifuge the solid, remove the supernatant, and place the solid in a 40°C vacuum drying oven to dry to constant weight to obtain p-benzene Form III of mesylate salt. After detection and analysis, it has an XRPD pattern as shown in FIG. 19.
⑨对苯甲磺酸盐晶型IV的制备⑨ Preparation of p-toluenesulfonate crystal form IV
将200mg对苯甲磺酸盐放入80℃下干燥4h即可得到对苯甲磺酸盐晶型IV，经检测分析，其有如图20所示的XRPD图。200 mg of p-toluenesulfonate was placed at 80° C. and dried for 4 hours to obtain p-toluenesulfonate crystal form IV, which has an XRPD pattern as shown in FIG. 20 after detection and analysis.
⑩硝酸盐的制备⑩Preparation of nitrate
称取50mg化合物实施例2自由碱加入1mL甲醇，室温搅拌不溶，向混悬液中加入157uL 1M的硝酸水溶液与1mL的乙醇混合液，在室温下搅拌12h后过滤，滤饼用2mL丙酮淋洗，最后固体放入40℃真空干燥箱中烘干至恒重，得到硝酸盐晶体。经检测分析，其有如图21所示的XRPD图。Weigh 50 mg of compound Example 2 Add 1 mL of methanol to the free base, stir at room temperature and insoluble, add 157uL 1M nitric acid aqueous solution and 1 mL of ethanol to the suspension, stir at room temperature for 12 hours, then filter, and rinse the filter cake with 2 mL of acetone Finally, the solid is placed in a vacuum drying oven at 40°C and dried to a constant weight to obtain nitrate crystals. After detection and analysis, it has an XRPD pattern as shown in FIG. 21.
3.稳定性实验3. Stability experiment
3.1实验目的：3.1 The purpose of the experiment:
考察候选化合物的自由碱或不同盐型在加速条件或影响因素条件下，化合物的物理化学稳定性，为盐型筛选与化合物盐贮存提供依据。Investigate the physicochemical stability of the free base or different salt types of candidate compounds under accelerated conditions or influencing factors, and provide a basis for salt type screening and compound salt storage.
3.2实验方案：3.2 Experimental program:
分别称取化合物实施例2自由碱晶型II和盐约2mg，密闭置60℃烘箱、敞口置室温RH95％(饱和KNO 3水溶液)和光照箱(5000lx±500lx)中，考察5天、10天，用HPLC，外标法测定盐的含量，并采用色谱峰面积归一化法计算盐有关物质的变化。 Weigh about 2 mg of the free base crystal form II and salt of compound example 2 respectively, and put them in a 60℃ oven, an open room with room temperature RH95% (saturated KNO 3 aqueous solution) and a light box (5000lx±500lx), and inspect for 5 days, 10 Day, HPLC and external standard method were used to determine the salt content, and the chromatographic peak area normalization method was used to calculate the change of salt related substances.
3.3实验结果：3.3 Experimental results:
3.3.1化合物实施例2自由碱晶型II和盐的物理化学稳定性结果如下表20所示，以下“-”表示未检测：3.3.1 The results of the physicochemical stability of the free base crystal form II and salt of Compound Example 2 are shown in Table 20 below. The following "-" means not detected:
表20：化合物实施例2自由碱和盐的稳定性实验结果Table 20: Compound Example 2 Free base and salt stability test results
Figure PCTCN2020082042-appb-000157
Figure PCTCN2020082042-appb-000157
以上数据显示：马来酸盐在高温及高湿条件下均较化合物实施例2自由碱晶型II稳定，且无明显杂质生成。The above data shows that the maleate is more stable than the free base crystal form II of Compound Example 2 under high temperature and high humidity conditions, and no obvious impurities are generated.
4.引湿性实验4. Hygroscopicity test
4.1实验目的：4.1 Experimental purpose:
考察化合物自由碱及盐在不同相对湿度条件下的引湿性，为化合物盐筛选与贮存提供依据。Investigate the hygroscopicity of free bases and salts of compounds under different relative humidity conditions to provide a basis for the screening and storage of compound salts.
4.2实验方案：4.2 Experimental program:
将化合物盐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。Put the compound salt in saturated water vapor with different relative humidity to make the compound and water vapor reach a dynamic equilibrium, and calculate the percentage of moisture absorption and weight gain of the compound after equilibrium.
4.3实验结果：4.3 Experimental results:
4.3.1化合物实施例2自由碱晶型II及其盐的引湿性4.3.1 Compound Example 2 The hygroscopicity of free base crystal form II and its salt
1)化合物实施例2自由碱晶型II在RH80％条件下吸湿增重约2.23％，有引湿性。1) Compound Example 2 Free base crystal form II at RH80% moisture absorption and weight gain of about 2.23%, with hygroscopicity.
2)马来酸盐晶型I在RH80％条件下吸湿增重1.36％，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，马来酸盐晶型I的XRPD谱图并未改变，即没有晶型转变。2) Maleate crystal form I absorbs moisture and increases weight by 1.36% under the condition of RH80%, which is slightly hygroscopic; after a cycle of moisture absorption and desorption under the condition of 0-95% relative humidity, maleate crystal form I The XRPD spectrum has not changed, that is, there is no crystal transformation.
5.溶解度实验5. Solubility experiment
5.1实验目的：5.1 Experiment purpose:
比较化合物自由碱及盐在水、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，为盐可成药性评估提供依据。Comparing the solubility of the compound free base and salt in water, artificial gastric juice (SGF), fasting artificial intestinal fluid (FaSSIF) and non-fasting artificial intestinal fluid (FeSSIF) and other media, provide a basis for the evaluation of salt's druggability.
5.2实验方案：5.2 Experimental program:
将约2mg化合物混悬到不同介质中24小时，用HPLC，外标法测定化合物室温下的热力学溶解度。About 2 mg of the compound was suspended in different media for 24 hours, and the thermodynamic solubility of the compound at room temperature was determined by HPLC and external standard method.
5.3实验结果：5.3 Experimental results:
5.3.1化合物实施例2自由碱晶型II、马来酸盐晶型I和对苯甲磺酸盐晶型IV的溶解度结果如下表21所示：5.3.1 Compound Example 2 The solubility results of free base crystal form II, maleate crystal form I and p-toluenesulfonate crystal form IV are shown in Table 21 below:
表21化合物实施例2自由碱和盐的晶型溶解度实验结果Table 21 Compound Example 2 Free base and salt crystal form solubility test results
Figure PCTCN2020082042-appb-000158
Figure PCTCN2020082042-appb-000158
从化合物实施例2自由碱晶型II及马来酸盐在四个媒介中溶解度结果可以看出，成盐后化合物仍在水中不溶，但成马来酸盐后提高了在FeSSIF介质中的溶解度；而成对苯甲磺酸盐后虽然提高了在介质FaSSIF中的溶解度，但是却降低了在SGF中的溶解度。From the results of compound example 2 free base crystal form II and the solubility of maleate in the four media, it can be seen that the compound is still insoluble in water after the salt is formed, but the solubility in FeSSIF medium is improved after the maleate is formed ; Although the p-toluenesulfonate salt increases the solubility in the medium FaSSIF, it reduces the solubility in SGF.
6.多晶筛选实验6. Polycrystalline screening experiment
6.1实验目的：6.1 Experiment purpose:
通过多晶筛选，找到比较稳定的晶型。Through polycrystalline screening, a relatively stable crystal form is found.
6.2实验方案：6.2 Experimental program:
选择有一定溶解度的有机溶剂、水，将化合物悬浮于溶剂体系中，室温搅拌打浆1周后，离心，弃掉上清液，固体在40℃条件真空干燥(-0.1Mpa)过夜后，测定固体的XRPD，并与原料化合物盐的XRPD比较。Choose an organic solvent and water with a certain degree of solubility, suspend the compound in the solvent system, stir and beat at room temperature for 1 week, centrifuge, discard the supernatant, and dry the solid under vacuum at 40℃ (-0.1Mpa) overnight, then determine the solid XRPD and compare with the XRPD of the starting compound salt.
6.3实验操作：6.3 Experimental operation:
1)化合物实施例2马来酸盐各种晶型的制备：1) Compound Example 2 Preparation of various crystal forms of maleate:
①马来酸盐晶型I的制备① Preparation of maleate crystal form I
称取500mg化合物实施例2自由碱晶型II，加入5mL N,N-二甲基甲酰胺超声未溶清，体系在50℃下加热至完全溶清，另称取183mg马来酸加1mL甲醇室温振荡完全溶清。在50℃下将马来酸的甲醇溶液加入碱的溶液中，很快析出沉淀，搅拌0.5h后向体系中加入10mL甲基叔丁基醚，继续搅拌1.5h后关闭加热，冷却至室温，最后过滤，滤饼用20mL甲基叔丁基醚淋洗，固体放入40℃真空干燥箱烘干至恒重，得到马来酸盐晶型I。Weigh 500 mg of compound Example 2 free base crystal form II, add 5 mL of N,N-dimethylformamide ultrasonically undissolved, heat the system at 50°C until it is completely dissolved, and weigh 183 mg of maleic acid plus 1 mL of methanol Shake at room temperature to dissolve completely. The methanol solution of maleic acid was added to the alkali solution at 50°C, and the precipitate precipitated out quickly. After stirring for 0.5h, 10mL methyl tert-butyl ether was added to the system. After stirring for 1.5h, the heating was turned off and the temperature was cooled to room temperature. Finally, it is filtered, the filter cake is rinsed with 20 mL of methyl tert-butyl ether, and the solid is placed in a vacuum drying oven at 40° C. and dried to a constant weight to obtain maleate crystal form I.
②马来酸盐晶型II的制备② Preparation of maleate crystal form II
称取498.37mg化合物实施例2自由碱晶型II，加入10mL甲醇，50℃搅拌不溶，为灰白色混悬液。另称取185.05mg马来酸加1mL甲醇超声溶清。将马来酸的甲醇溶液加入50℃的自由碱混悬液中，混悬液立即由灰白色变成黄色。在50℃下搅拌反应2h后关闭加热，将至室温，最后过滤，滤饼用10mL甲基叔丁基醚淋洗。固体放入40℃真空干燥箱烘干至恒重，得到马来酸盐晶型II。Weigh 498.37 mg of the free base crystal form II of Compound Example 2 and add 10 mL of methanol, stir insoluble at 50°C, and become an off-white suspension. Separately weigh 185.05 mg maleic acid and add 1 mL methanol to ultrasonically dissolve it. The methanol solution of maleic acid was added to the free base suspension at 50°C, and the suspension immediately changed from off-white to yellow. After stirring the reaction at 50°C for 2 hours, the heating was turned off, the temperature was brought to room temperature, and finally filtered, and the filter cake was rinsed with 10 mL of methyl tert-butyl ether. The solid was dried in a vacuum drying oven at 40°C to a constant weight to obtain maleate crystal form II.
③马来酸盐晶型III的制备③ Preparation of maleate crystal form III
称取30mg马来酸盐晶型I，加入200uL四氢呋喃，在50℃下打浆4d，最后将固体离心，去除上清液后固体放入40℃真空干燥箱中烘干至恒重，得到马 来酸盐晶型III。Weigh 30 mg of maleate crystal form I, add 200 uL of tetrahydrofuran, and beat the slurry at 50°C for 4 days. Finally, the solid is centrifuged, the supernatant is removed, and the solid is dried in a vacuum drying oven at 40°C to a constant weight. Salt crystal form III.
④马来酸盐晶型IV的制备④ Preparation of maleate crystal form IV
称取50.23mg马来酸盐晶型I，加入0.75mL N-甲基吡咯烷酮在50℃下完全溶清，向体系中加入1mL异丙醚，很快析出黄色固体，在50℃下反应2h后继续加入2mL异丙醚，并关闭加热将至室温，最后将固体离心，去除上清液后固体放入40℃真空干燥箱中烘干至恒重，得到马来酸盐晶型IV。Weigh 50.23mg maleate crystal form I, add 0.75mL N-methylpyrrolidone to completely dissolve it at 50℃, add 1mL isopropyl ether to the system, and quickly precipitate a yellow solid. After reacting at 50℃ for 2h Continue to add 2 mL of isopropyl ether and turn off the heating to bring to room temperature. Finally, the solid is centrifuged, and the supernatant is removed, and the solid is placed in a vacuum drying oven at 40° C. and dried to a constant weight to obtain maleate crystal form IV.
6.4实验结果：6.4 Experimental results:
6.4.1化合物实施例2马来酸盐各晶型之间的竞争关系：6.4.1 Competitive relationship among the crystalline forms of the maleate salt of Compound Example 2:
1)马来酸盐晶型II在甲醇中很稳定。1) Maleate crystal form II is very stable in methanol.
2)马来酸盐晶型III在THF中稳定。2) Maleate salt form III is stable in THF.
3)马来酸盐晶型I在丙酮、乙腈、乙醇、乙酸乙酯、异丙醇和水中较稳定。3) Maleate salt crystal form I is relatively stable in acetone, acetonitrile, ethanol, ethyl acetate, isopropanol and water.
7.PK实验7. PK experiment
7.1实验目的：7.1 Experiment purpose:
以SD大鼠，雄性为受试动物，研究化合物实施例2自由碱晶型II、马来酸盐晶型I及对苯甲磺酸盐晶型IV口服给药在大鼠体内(血浆)的药代动力学行为。Taking SD rats and males as the test animals, study compound Example 2 Free base crystal form II, maleate crystal form I and p-toluenesulfonate crystal form IV were administered orally in rats (plasma) Pharmacokinetic behavior.
7.2实验方案：7.2 Experimental program:
将化合物实施例2自由碱晶型II、马来酸盐晶型I及对苯甲磺酸盐晶型IV，用含0.5％的HPMC K4M的水溶液水溶液混悬均匀后，灌胃，大鼠给药，平行三只大鼠，给药剂量为100mg/kg，化合物的量全部折算成相同化合物实施例2自由碱的量。Compound Example 2 free base crystalline form II, maleate crystalline form I and p-toluenesulfonate crystalline form IV were uniformly suspended in an aqueous solution containing 0.5% HPMC K4M, and the rats were given by intragastric administration. Medicine, three rats in parallel, the dose of 100mg/kg, the amount of the compound is all converted to the amount of the free base of the same compound in Example 2.
7.3实验结果：7.3 Experimental results:
化合物实施例2自由碱及不同盐的晶型PK实验结果如下表22所示：The PK experiment results of the crystalline form of the free base and different salts of Compound Example 2 are shown in Table 22 below:
表22：化合物实施例2自由碱和盐的晶型SD大鼠PK实验结果Table 22: Compound Example 2 Free base and salt crystal form SD rat PK experiment results
Figure PCTCN2020082042-appb-000159
Figure PCTCN2020082042-appb-000159
由表22可知：It can be seen from Table 22:
1)对比化合物实施例2自由碱晶型II的暴露量，马来酸盐晶型I及对苯甲磺酸盐晶型IV有所提高。1) The exposure amount of the free base crystal form II of Comparative Compound Example 2, maleate crystal form I and p-toluenesulfonate crystal form IV have increased.
2)对比化合物实施例2自由碱晶型II的T max，马来酸盐晶型I的T max有所减小。 2) Comparative compound example 2 has a free base crystal form II T max , and maleate salt crystal form I has a reduced T max .
3)化合物实施例2自由碱晶型II、马来酸盐晶型I及对苯甲磺酸盐晶型IV，24h内大鼠体内血浆中药物的浓度的变化情况如图22所示。3) Compound Example 2 Free base crystal form II, maleate crystal form I and p-toluenesulfonate crystal form IV, the changes of the concentration of the drug in the plasma in rats within 24 hours are shown in FIG. 22.

Claims (31)

  1. 一种通式(Ia)所示的化合物，其结构如下：A compound represented by general formula (Ia), its structure is as follows:
    Figure PCTCN2020082042-appb-100001
    Figure PCTCN2020082042-appb-100001
    其中：among them:
    W选自-N-或-CH-；W is selected from -N- or -CH-;
    R 1选自氢原子、卤素、氨基、硝基、羟基、氰基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6卤代烷氧基、C 1-6羟烷基、C 3-8环烷基、-(CH 2) nR 3、-(CH 2) nOR 3、-(CH 2) nC(O)R 3、-(CH 2) nNR 3R 4或-(CH 2) nC(O)NR 3R 4； R 1 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R 3 , -(CH 2 ) n OR 3 , -(CH 2 ) n C(O)R 3 , -(CH 2 ) n NR 3 R 4 or -(CH 2 ) n C(O)NR 3 R 4 ;
    R 2选自氢原子、卤素、氨基、硝基、羟基、氰基、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卤代烷氧基、C 3-8环烷基或3-8元杂环基； R 2 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl or 3-8 membered heterocyclic group;
    R 3和R 4相同或不同，且各自独立地选自氢原子、氘原子、卤素、氰基、羟基、氨基、硝基、C 1-6烷基、C 1-6卤代烷基、C 3-8环烷基或3-8元杂环基，其中所述的C 1-6烷基、C 1-6卤代烷基、氨基、C 3-8环烷基和3-8元杂环基任选进一步被选自氘原子、C 1-6烷基、C 1-6卤代烷基、卤素、羟基、氨基、硝基、氰基、C 1-6烷氧基、C 1-6羟烷基、C 3-8环烷基和3-6元杂环基中的一个或多个取代基所取代； R 3 and R 4 are the same or different, and are each independently selected from hydrogen atom, deuterium atom, halogen, cyano, hydroxyl, amino, nitro, C 1-6 alkyl, C 1-6 haloalkyl, C 3- 8 cycloalkyl or 3-8 membered heterocyclic group, wherein the C 1-6 alkyl, C 1-6 haloalkyl, amino, C 3-8 cycloalkyl and 3-8 membered heterocyclic group are optional Further selected from deuterium atom, C 1-6 alkyl, C 1-6 haloalkyl, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl and 3-6 membered heterocyclic group substituted by one or more substituents;
    M为无机酸或有机酸，其中所述无机酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸或磷酸；有机酸选自2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；M is an inorganic acid or an organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1- Hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-amino Benzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid , Lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, Galactonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid , Malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid Acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid ；
    y为1、2或3的整数；y is an integer of 1, 2 or 3;
    n为0、1、2或3的整数；且n is an integer of 0, 1, 2 or 3; and
    t为0、1、2、3、4、5或6的整数。t is an integer of 0, 1, 2, 3, 4, 5, or 6.
  2. 根据权利要求1所述的通式(Ia)所示的化合物，其特征在于，化合物结构如式(IIa)所示：The compound represented by general formula (Ia) according to claim 1, wherein the structure of the compound is represented by formula (IIa):
    Figure PCTCN2020082042-appb-100002
    Figure PCTCN2020082042-appb-100002
  3. 根据权利要求1所述的通式(Ia)所示的化合物，其特征在于，所述化合物结构如下：The compound represented by general formula (Ia) according to claim 1, wherein the structure of the compound is as follows:
    Figure PCTCN2020082042-appb-100003
    Figure PCTCN2020082042-appb-100003
    Figure PCTCN2020082042-appb-100004
    Figure PCTCN2020082042-appb-100004
  4. 根据权利要求1所述的通式(Ia)所示的化合物，其特征在于，所述化合物结构如式(IIIa)所示：The compound represented by formula (Ia) according to claim 1, wherein the structure of the compound is represented by formula (IIIa):
    Figure PCTCN2020082042-appb-100005
    Figure PCTCN2020082042-appb-100005
  5. 根据权利要求4所述的通式(Ia)所示的化合物，其特征在于，所述的式(IIIa)化合物为无水物，进一步还包含水，所述水为管道水或结晶水，优选地，含有0.5-8个水分子，更优选0.5-4个水分子，进一步优选0.5-2.5个水分子，更进一步优选1个水分子。The compound represented by general formula (Ia) according to claim 4, wherein the compound of formula (IIIa) is an anhydrate, and further comprises water, and the water is pipeline water or crystal water, preferably It contains 0.5-8 water molecules, more preferably 0.5-4 water molecules, still more preferably 0.5-2.5 water molecules, and still more preferably 1 water molecule.
  6. 根据权利要求4所述的通式(Ia)所示的化合物，其特征在于，所述的式(IIIa)化合物为无定型。The compound represented by the general formula (Ia) according to claim 4, wherein the compound of the formula (IIIa) is amorphous.
  7. 根据权利要求1所述的通式(Ia)所示的化合物，其特征在于，化合物结构如式(IVa)所示：The compound represented by general formula (Ia) according to claim 1, wherein the structure of the compound is represented by formula (IVa):
    Figure PCTCN2020082042-appb-100006
    Figure PCTCN2020082042-appb-100006
  8. 一种通式(I)所示化合物的晶型，其结构如式(I)所示：A crystal form of the compound represented by general formula (I), and its structure is as shown in formula (I):
    Figure PCTCN2020082042-appb-100007
    Figure PCTCN2020082042-appb-100007
    其中：among them:
    W选自N或CH；W is selected from N or CH;
    R 1选自氢原子、卤素、氨基、硝基、羟基、氰基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6卤代烷氧基、C 1-6羟烷基、C 3-8环烷基、-(CH 2) nR 3、-(CH 2) nOR 3、-(CH 2) nC(O)R 3、-(CH 2) nNR 3R 4或-(CH 2) nC(O)NR 3R 4； R 1 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R 3 , -(CH 2 ) n OR 3 , -(CH 2 ) n C(O)R 3 , -(CH 2 ) n NR 3 R 4 or -(CH 2 ) n C(O)NR 3 R 4 ;
    R 2选自氢原子、卤素、氨基、硝基、羟基、氰基、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卤代烷氧基、C 3-8环烷基或3-8元杂环基； R 2 is selected from hydrogen atom, halogen, amino, nitro, hydroxyl, cyano, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl or 3-8 membered heterocyclic group;
    R 3和R 4相同或不同，且各自独立地选自氢原子、氘原子、卤素、氰基、羟 基、氨基、硝基、C 1-6烷基、C 1-6卤代烷基、C 3-8环烷基或3-8元杂环基，其中所述的C 1-6烷基、C 1-6卤代烷基、氨基、C 3-8环烷基和3-8元杂环基任选进一步被选自氘原子、C 1-6烷基、C 1-6卤代烷基、卤素、羟基、氨基、硝基、氰基、C 1-6烷氧基、C 1-6羟烷基、C 3-8环烷基和3-6元杂环基中的一个或多个取代基所取代； R 3 and R 4 are the same or different, and are each independently selected from hydrogen atom, deuterium atom, halogen, cyano, hydroxyl, amino, nitro, C 1-6 alkyl, C 1-6 haloalkyl, C 3- 8 cycloalkyl or 3-8 membered heterocyclic group, wherein the C 1-6 alkyl, C 1-6 haloalkyl, amino, C 3-8 cycloalkyl and 3-8 membered heterocyclic group are optional Further selected from deuterium atom, C 1-6 alkyl, C 1-6 haloalkyl, halogen, hydroxyl, amino, nitro, cyano, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl and 3-6 membered heterocyclic group substituted by one or more substituents;
    M为无机酸或有机酸，其中所述无机酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸或磷酸；有机酸选自2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；M is an inorganic acid or an organic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; the organic acid is selected from 2,5-dihydroxybenzoic acid, 1- Hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-amino Benzoic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid , Lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, Galactonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid , Malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid Acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid ；
    x选自0、1、2或3的整数；x is selected from an integer of 0, 1, 2 or 3;
    n为0、1、2或3的整数；且n is an integer of 0, 1, 2 or 3; and
    t为0、1、2、3、4、5或6的整数。t is an integer of 0, 1, 2, 3, 4, 5, or 6.
  9. 根据权利要求8所述的通式(I)所示化合物的晶型，其特征在于，所述化合物结构如式(II)所示：The crystal form of the compound represented by general formula (I) according to claim 8, wherein the structure of the compound is represented by formula (II):
    Figure PCTCN2020082042-appb-100008
    Figure PCTCN2020082042-appb-100008
  10. 根据权利要求8所述的通式(I)所示化合物的晶型，所述化合物结构如下：The crystal form of the compound represented by the general formula (I) according to claim 8, wherein the structure of the compound is as follows:
    Figure PCTCN2020082042-appb-100009
    Figure PCTCN2020082042-appb-100009
    Figure PCTCN2020082042-appb-100010
    Figure PCTCN2020082042-appb-100010
  11. 根据权利要求8所述的通式(I)所示化合物的晶型，其特征在于，化合物结构如式(III)所示：The crystalline form of the compound represented by the general formula (I) according to claim 8, wherein the compound structure is represented by the formula (III):
    Figure PCTCN2020082042-appb-100011
    Figure PCTCN2020082042-appb-100011
  12. 根据权利要求11所述的通式(I)所示化合物的晶型，其特征在于，x为0，所述的式(III)化合物为自由碱的晶型；The crystalline form of the compound represented by general formula (I) according to claim 11, wherein x is 0, and the compound of formula (III) is a free base crystalline form;
    或在x选自1、2或3时，所述的式(III)化合物为盐晶型；优选x为1。Or when x is selected from 1, 2 or 3, the compound of formula (III) is a salt crystal form; preferably x is 1.
  13. 根据权利要求11或12所述的通式(I)所示化合物的晶型，其特征在于，所述的式(III)化合物为无水物，进一步还包含水，所述水为管道水或结晶水，优选地，含有0.5-8个水分子，更优选0.5-4个水分子，进一步优选0.5-2.5个水分子，更进一步优选1个水分子。The crystal form of the compound represented by general formula (I) according to claim 11 or 12, wherein the compound of formula (III) is an anhydrate, and further comprises water, and the water is pipeline water or Crystal water, preferably, contains 0.5-8 water molecules, more preferably 0.5-4 water molecules, still more preferably 0.5-2.5 water molecules, even more preferably 1 water molecule.
  14. 根据权利要求8所述的通式(I)所示化合物的晶型，其特征在于，所述化合物结构如式(IV)所示：The crystal form of the compound represented by general formula (I) according to claim 8, wherein the structure of the compound is represented by formula (IV):
    Figure PCTCN2020082042-appb-100012
    Figure PCTCN2020082042-appb-100012
  15. 一种制备权利要求1-7任意一项所述化合物或权利要求8-14任意一项所述化合物的晶型的方法，具体包括如下步骤：A method for preparing the compound described in any one of claims 1-7 or the crystal form of the compound described in any one of claims 8-14, specifically comprising the following steps:
    1)称取适量的自由碱，用良性溶剂溶解；1) Weigh an appropriate amount of free base and dissolve it with a benign solvent;
    2)任选地，称取适量的反离子酸，用有机溶剂溶解；优选地，反离子酸的量1.2当量；2) Optionally, weigh an appropriate amount of counter ion acid and dissolve it with an organic solvent; preferably, the amount of counter ion acid is 1.2 equivalents;
    3)任选地，把上述两种溶液合并，搅拌后若无沉淀析出，滴加不良溶剂直至出现浑浊；3) Optionally, combine the above two solutions, and if there is no precipitation after stirring, add dropwise poor solvent until turbidity appears;
    4)搅拌、析晶得到目标产物；4) Stir and crystallize to obtain the target product;
    其中：among them:
    所述的良性溶剂选自甲醇、四氢呋喃、N,N-二甲基甲酰胺、N，N-二甲基乙酰胺或N-甲基吡咯烷酮；优选N,N-二甲基甲酰胺和N-甲基吡咯烷酮；The benign solvent is selected from methanol, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidone; preferably N,N-dimethylformamide and N- Methylpyrrolidone;
    所述的有机溶剂选自甲醇、乙醇、乙酸乙酯、二氯甲烷、丙酮、正己烷、石 油醚、苯、甲苯、氯仿、乙腈、四氯化碳、二氯乙烷、四氢呋喃、2-丁酮、3-戊酮、庚烷、甲基叔丁基醚、异丙醚、1,4-二氧六环、叔丁醇或N，N-二甲基甲酰胺；优选甲醇和乙醇；上述良性溶剂和有机溶液使用时需互溶；The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol and ethanol; the above The benign solvent and organic solution must be miscible when used;
    所述的不良溶剂选自庚烷、水、甲基叔丁基醚、甲苯、异丙醚、乙酸乙酯、丙酮或乙腈；优选乙酸乙酯、甲基叔丁基醚、异丙醚和乙腈；The poor solvent is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone or acetonitrile; preferably ethyl acetate, methyl tert-butyl ether, isopropyl ether and acetonitrile ；
    所述的反离子酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸、磷酸、2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；优选甲磺酸、硫酸、盐酸、硝酸、苯磺酸、马来酸、己二酸、对甲基苯磺酸、柠檬酸、丙二酸和L-苹果酸；更优选马来酸。The counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroglutarate Amino acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid , 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphor acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid Acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably methanesulfonic acid, sulfuric acid, hydrochloric acid, nitric acid , Benzenesulfonic acid, maleic acid, adipic acid, p-toluenesulfonic acid, citric acid, malonic acid, and L-malic acid; maleic acid is more preferred.
  16. 一种制备根据权利要求1-7任意一项所述化合物或权利要求8-14任意一项所述化合物的晶型的方法，具体包括如下步骤：A method for preparing the compound according to any one of claims 1-7 or the crystal form of the compound according to any one of claims 8-14, specifically comprising the following steps:
    1)称取适量的自由碱，用不良性溶剂混悬；1) Weigh an appropriate amount of free base and suspend it in a poor solvent;
    2)任选地，称取适量的反离子酸，用有机溶剂溶解；优选地，反离子酸的量1.2当量；2) Optionally, weigh an appropriate amount of counter ion acid and dissolve it with an organic solvent; preferably, the amount of counter ion acid is 1.2 equivalents;
    3)任选地，将2)中溶液加入1)中混悬液；3) Optionally, add the solution in 2) to the suspension in 1);
    4)搅拌、析晶得到目标产物；4) Stir and crystallize to obtain the target product;
    其中：among them:
    所述的不良性溶剂选自丙酮、乙酸乙酯、乙腈、乙醇、88％丙酮、四氢呋喃、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、叔丁醇或2-丁酮；优选甲醇、乙醇、四氢呋喃、乙酸乙酯、乙腈或丙酮；优选甲醇、乙醇、四氢呋喃、乙酸乙酯、乙腈和丙酮；The poor solvent is selected from acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N , N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol or 2-butanone; preferably methanol, ethanol, tetrahydrofuran, ethyl acetate, acetonitrile or acetone; preferably methanol, Ethanol, tetrahydrofuran, ethyl acetate, acetonitrile and acetone;
    所述的有机溶剂选自甲醇、乙醇、乙酸乙酯、二氯甲烷、丙酮、正己烷、石油醚、苯、甲苯、氯仿、乙腈、四氯化碳、二氯乙烷、四氢呋喃、2-丁酮、3-戊酮、庚烷、甲基叔丁基醚、异丙醚、1,4-二氧六环、叔丁醇或N，N-二甲基甲酰胺；优选甲醇和乙醇；上述良性溶剂和有机溶液使用时需互溶；The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol and ethanol; the above The benign solvent and organic solution must be miscible when used;
    所述的反离子酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸、磷酸、2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；优选马来酸。The counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroglutarate Amino acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid , 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphor acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid Acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably maleic acid.
  17. 一种制备权利要求1-7任意一项所述化合物或权利要求8-14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 compound described in any one of claims 1-7 or the crystal form of the compound described in any one of claims 8-14, specifically comprising the following steps:
    1)称取适量的自由碱，加入良性溶剂，加热至溶解；1) Weigh an appropriate amount of free base, add a benign solvent, and heat to dissolve;
    2)任选地，称取适量的反离子酸，用有机溶剂溶解；优选地，反离子酸的量1.2当量；2) Optionally, weigh an appropriate amount of counter ion acid and dissolve it with an organic solvent; preferably, the amount of counter ion acid is 1.2 equivalents;
    3)搅拌、降温析晶得到目标产物；3) Stirring, cooling and crystallization to obtain the target product;
    其中：among them:
    所述的良性溶剂选自甲醇、四氢呋喃、N,N-二甲基甲酰胺、N，N-二甲基乙酰胺或N-甲基吡咯烷酮；优选N,N-二甲基甲酰胺和N-甲基吡咯烷酮；The benign solvent is selected from methanol, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidone; preferably N,N-dimethylformamide and N- Methylpyrrolidone;
    所述的有机溶剂选自甲醇、乙醇、乙酸乙酯、二氯甲烷、丙酮、正己烷、石油醚、苯、甲苯、氯仿、乙腈、四氯化碳、二氯乙烷、四氢呋喃、2-丁酮、3-戊酮、庚烷、甲基叔丁基醚、异丙醚、1,4-二氧六环、叔丁醇或N，N-二甲基甲酰胺；优选甲醇和乙醇；上述良性溶剂和有机溶液使用时需互溶；The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butane Ketone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol or N,N-dimethylformamide; preferably methanol and ethanol; the above The benign solvent and organic solution must be miscible when used;
    所述的反离子酸选自盐酸、硫酸、硝酸、氢溴酸、氢氟酸、氢碘酸、磷酸、2,5-二羟基苯甲酸、1-羟基-2-萘甲酸、醋酸、二氯醋酸、三氯醋酸、乙酰氧肟酸、己二酸、苯磺酸、4-氯苯磺酸、苯甲酸、4-乙酰氨基苯甲酸、4-氨基苯甲酸、癸酸、己酸、辛酸、肉桂酸、柠檬酸、环己烷氨基磺酸、樟脑磺酸、天门冬氨酸、樟脑酸、葡萄糖酸、葡糖醛酸、谷氨酸、异抗坏血酸、乳酸、苹果酸、扁桃酸、焦谷氨酸、酒石酸、十二烷基硫酸、二苯甲酰酒石酸、乙烷-1,2-二磺酸、乙磺酸、蚁酸、富马酸、半乳糖酸、龙胆酸、戊二酸、2-酮戊二酸、乙醇酸、马尿酸、羟乙基磺酸、乳糖酸、抗坏血酸、天冬氨酸、月桂酸、樟脑酸、马来酸、丙二酸、甲磺酸、1,5-萘二磺酸、萘-2-磺酸、烟酸、油酸、乳清酸、草酸、棕榈酸、双羟萘酸、丙酸、水杨酸、4-氨基水杨酸、癸二酸、硬脂酸、丁二酸、硫氰酸、十一 碳烯酸、三氟乙酸、苯磺酸、对甲基苯磺酸或L-苹果酸；优选甲磺酸、硫酸、盐酸、硝酸、苯磺酸、马来酸、己二酸、对甲基苯磺酸、柠檬酸、丙二酸和L-苹果酸；更优选马来酸。The counter ion acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloro Acetic acid, trichloroacetic acid, acetohydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylaminobenzoic acid, 4-aminobenzoic acid, capric acid, caproic acid, caprylic acid, Cinnamic acid, citric acid, cyclohexane sulfamic acid, camphor sulfonic acid, aspartic acid, camphor acid, gluconic acid, glucuronic acid, glutamic acid, erythorbic acid, lactic acid, malic acid, mandelic acid, pyroglutarate Amino acid, tartaric acid, lauryl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactonic acid, gentisic acid, glutaric acid , 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphor acid, maleic acid, malonic acid, methanesulfonic acid, 1, 5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, niacin, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid Acid, stearic acid, succinic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid or L-malic acid; preferably methanesulfonic acid, sulfuric acid, hydrochloric acid, nitric acid , Benzenesulfonic acid, maleic acid, adipic acid, p-toluenesulfonic acid, citric acid, malonic acid, and L-malic acid; maleic acid is more preferred.
  18. 根据权利要求1-7任意一项所述的通式(Ia)所示的化合物，其特征在于：M选自马来酸、对苯甲磺酸、盐酸、硝酸、水杨酸、甲磺酸、苯磺酸或1,5-萘二磺酸；优选马来酸；The compound represented by general formula (Ia) according to any one of claims 1-7, characterized in that: M is selected from maleic acid, p-toluenesulfonic acid, hydrochloric acid, nitric acid, salicylic acid, methanesulfonic acid , Benzenesulfonic acid or 1,5-naphthalenedisulfonic acid; preferably maleic acid;
    y为1。y is 1.
  19. 根据权利要求8-14任意一项所述的通式(I)所示化合物的晶型，其特征在于，The crystal form of the compound represented by general formula (I) according to any one of claims 8-14, characterized in that:
    M选自马来酸、对苯甲磺酸、盐酸、硝酸、水杨酸、甲磺酸、苯磺酸或1,5-萘二磺酸，优选马来酸；M is selected from maleic acid, p-toluenesulfonic acid, hydrochloric acid, nitric acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid or 1,5-naphthalenedisulfonic acid, preferably maleic acid;
    x为1。x is 1.
  20. 式(IV)所示化合物的晶型，其自由碱结晶形式I，X-射线粉末衍射图谱在2θ为6.4和26.7处(2θ±0.2°)具有特征峰，优选地，还包含在2θ±0.2°为7.2、13.1、16.8、17.7、18.9、20.2、21.2和28.2处(2θ±0.2°)具有特征峰；The crystalline form of the compound represented by formula (IV), its free base crystalline form I, X-ray powder diffraction pattern has characteristic peaks at 2θ of 6.4 and 26.7 (2θ±0.2°), preferably, it is also contained in 2θ±0.2 ° 7.2, 13.1, 16.8, 17.7, 18.9, 20.2, 21.2 and 28.2 (2θ±0.2°) have characteristic peaks;
    其自由碱结晶形式II，X-射线粉末衍射图谱在2θ为8.5、13.4和17.0处具有特征峰，优选地，在2θ为10.2、10.9、12.8、13.1、16.1、17.8、18.8、19.5、23.0、23.9、24.4、25.5、26.1和27.6处(2θ±0.2°)具有特征峰。Its free base crystalline form II, the X-ray powder diffraction pattern has characteristic peaks at 2θ of 8.5, 13.4 and 17.0, preferably at 2θ of 10.2, 10.9, 12.8, 13.1, 16.1, 17.8, 18.8, 19.5, 23.0, There are characteristic peaks at 23.9, 24.4, 25.5, 26.1 and 27.6 (2θ±0.2°).
  21. 式(IV)所示化合物的晶型，其为马来酸盐晶型，且x为1，The crystal form of the compound represented by formula (IV), which is the maleate crystal form, and x is 1,
    其马来酸盐晶型I，X-射线粉末衍射图谱在2θ为5.5和16.3处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为10.8、15.3、17.7、26.1、26.4和27.0处(2θ±0.2°)具有特征峰，更进一步在2θ为13.0、14.9、20.0和20.8处(2θ±0.2°)具有特征峰；Its maleate salt crystal form I, X-ray powder diffraction pattern has characteristic peaks at 2θ of 5.5 and 16.3 (2θ±0.2°); preferably, it is further contained in 2θ of 10.8, 15.3, 17.7, 26.1, 26.4 There are characteristic peaks at and 27.0 (2θ±0.2°), and furthermore, there are characteristic peaks at 2θ at 13.0, 14.9, 20.0 and 20.8 (2θ±0.2°);
    其马来酸盐晶型II，X-射线粉末衍射图谱在2θ为5.5、10.8、16.3和17.6处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为14.0、26.4、26.6和27.2的特征峰，更进一步包含在2θ为8.8、12.9、18.4、18.9、20.6、21.5、22.4和22.8处(2θ±0.2°)具有特征峰；Its maleate salt crystal form II, the X-ray powder diffraction pattern has characteristic peaks at 2θ of 5.5, 10.8, 16.3, and 17.6 (2θ±0.2°); preferably, it is further contained in 2θ of 14.0, 26.4, 26.6 The characteristic peaks of and 27.2 further include characteristic peaks at 2θ of 8.8, 12.9, 18.4, 18.9, 20.6, 21.5, 22.4 and 22.8 (2θ±0.2°);
    其马来酸盐晶型III，X-射线粉末衍射图谱在2θ为4.9、15.1、17.2、17.5、26.5和26.9处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为9.8、14.7、18.4和19.8处(2θ±0.2°)具有特征峰，更进一步包含在2θ为12.3、13.0、14.0和27.5处(2θ±0.2°)具有特征峰；Its maleate salt crystal form III, X-ray powder diffraction pattern has characteristic peaks at 2θ of 4.9, 15.1, 17.2, 17.5, 26.5 and 26.9 (2θ±0.2°); preferably, it is further contained in 2θ of 9.8 There are characteristic peaks at, 14.7, 18.4 and 19.8 (2θ±0.2°), and further includes characteristic peaks at 2θ 12.3, 13.0, 14.0 and 27.5 (2θ±0.2°);
    其马来酸盐晶型IV，X-射线粉末衍射图谱在2θ为具有4.8、16.7的特征峰；优选地，进一步还包含在2θ为9.6、11.9和14.9处(2θ±0.2°)具有特征峰，更进一步包含在2θ为7.2、12.9、14.3、19.1、19.3、21.5和23.9处(2θ±0.2°)具有特征峰。The maleate salt crystal form IV, the X-ray powder diffraction pattern has characteristic peaks of 4.8 and 16.7 at 2θ; preferably, it further includes characteristic peaks at 2θ of 9.6, 11.9 and 14.9 (2θ±0.2°) , And further include characteristic peaks at 2θ of 7.2, 12.9, 14.3, 19.1, 19.3, 21.5, and 23.9 (2θ±0.2°).
  22. 式(IV)所示化合物的晶型，其为盐酸盐晶型，且x为1，X-射线粉末衍射图谱在2θ为5.8和17.2处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为12.0、13.6、16.4、21.7、23.0、26.1、26.3和27.1处(2θ±0.2°)具有特征峰，更进一步包含在2θ为8.9、28.8和30.1处(2θ±0.2°)具有特征峰。The crystal form of the compound represented by formula (IV) is the hydrochloride crystal form, and x is 1, and the X-ray powder diffraction pattern has characteristic peaks at 2θ of 5.8 and 17.2 (2θ±0.2°); preferably, It further includes characteristic peaks at 2θ of 12.0, 13.6, 16.4, 21.7, 23.0, 26.1, 26.3 and 27.1 (2θ±0.2°), and further includes 2θ at 8.9, 28.8 and 30.1 (2θ±0.2°) With characteristic peaks.
  23. 式(IV)所示化合物的晶型，其为对苯甲磺酸盐的晶型，且x为1，其对苯甲磺酸盐晶型I，X-射线粉末衍射图谱在2θ为6.1、11.3、17.0、17.5和18.3处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为12.7、19.1、19.9、20.6和22.2处(2θ±0.2°)具有特征峰；The crystal form of the compound represented by formula (IV), which is the crystal form of p-toluenesulfonate, and x is 1, its p-toluenesulfonate crystal form I, X-ray powder diffraction pattern at 2θ is 6.1, 11.3, 17.0, 17.5, and 18.3 have characteristic peaks (2θ±0.2°); preferably, it further includes characteristic peaks at 2θ of 12.7, 19.1, 19.9, 20.6 and 22.2 (2θ±0.2°);
    其对苯甲磺酸盐晶型II，X-射线粉末衍射图谱在2θ为5.4、8.6、11.4、16.8、18.2、19.7、20.4和21.9处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ±0.2为13.5、13.7、15.7、17.2、23.7、25.5和27.6处(2θ±0.2°)具有特征峰，更进一步包含在2θ为16.0、21.4、25.9和28.7处(2θ±0.2°)具有特征峰；Its p-toluenesulfonate crystal form II, X-ray powder diffraction pattern has characteristic peaks at 2θ of 5.4, 8.6, 11.4, 16.8, 18.2, 19.7, 20.4 and 21.9 (2θ±0.2°); preferably, further It also includes characteristic peaks at 2θ±0.2 at 13.5, 13.7, 15.7, 17.2, 23.7, 25.5 and 27.6 (2θ±0.2°), and further includes 2θ at 16.0, 21.4, 25.9, and 28.7 (2θ±0.2°). ) Has a characteristic peak;
    其对苯甲磺酸盐晶型III，X-射线粉末衍射图谱在2θ为4.9、8.6、13.2、18.9、20.6和25.2处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为9.6、10.9、12.6、15.0、15.6、17.0、22.6、25.8和27.5处(2θ±0.2°)具有特征峰，更进一步包含在2θ为10.5、13.9、16.5、17.7、21.7、26.1、26.6、26.9、27.8、29.8和32.2处(2θ±0.2°)具有特征峰；Its p-toluenesulfonate crystal form III, X-ray powder diffraction pattern has characteristic peaks at 2θ of 4.9, 8.6, 13.2, 18.9, 20.6 and 25.2 (2θ±0.2°); preferably, it is further contained in 2θ There are characteristic peaks at 9.6, 10.9, 12.6, 15.0, 15.6, 17.0, 22.6, 25.8 and 27.5 (2θ±0.2°), which are further included in 2θ at 10.5, 13.9, 16.5, 17.7, 21.7, 26.1, 26.6, 26.9 , 27.8, 29.8 and 32.2 (2θ±0.2°) have characteristic peaks;
    其对苯甲磺酸盐晶型IV，X-射线粉末衍射图谱在2θ为5.5、11.9、16.3、19.4和25.5处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为13.8、18.1、18.6、20.1、21.3、22.9和26.5处(2θ±0.2°)具有特征峰，更进一步包含8.7、9.1、10.2、10.8、12.9和20.4处(2θ±0.2°)具有特征峰。Its p-toluenesulfonate crystal form IV, the X-ray powder diffraction pattern has characteristic peaks at 2θ of 5.5, 11.9, 16.3, 19.4 and 25.5 (2θ±0.2°); preferably, it is further contained at 2θ of 13.8 , 18.1, 18.6, 20.1, 21.3, 22.9 and 26.5 (2θ±0.2°) have characteristic peaks, further including 8.7, 9.1, 10.2, 10.8, 12.9 and 20.4 (2θ±0.2°) have characteristic peaks.
  24. 式(IV)所示化合物的晶型，其为硝酸盐晶型，且x为1，X-射线粉末衍射图谱在2θ为5.0、16.3、16.7和28.0处(2θ±0.2°)具有特征峰；优选地，进一步还包含在2θ为8.2、8.5、11.8、13.2和29.1处(2θ±0.2°)具有特征峰，更进一步包含在2θ为19.7、20.4、21.0和24.5处(2θ±0.2°)具有特征峰。The crystal form of the compound represented by formula (IV), which is a nitrate crystal form, and x is 1, and the X-ray powder diffraction pattern has characteristic peaks at 2θ of 5.0, 16.3, 16.7 and 28.0 (2θ±0.2°); Preferably, it further includes characteristic peaks at 2θ of 8.2, 8.5, 11.8, 13.2, and 29.1 (2θ±0.2°), and further includes having characteristic peaks at 2θ of 19.7, 20.4, 21.0, and 24.5 (2θ±0.2°) Characteristic peaks.
  25. 一种药物组合物，其含有治疗有效量的权利要求1～7中任一项所述的通式(Ia)所示化合物，以及一种或多种药学上可接受的载体。A pharmaceutical composition containing a therapeutically effective amount of the compound represented by the general formula (Ia) according to any one of claims 1 to 7, and one or more pharmaceutically acceptable carriers.
  26. 根据权利要求1～7任意一项所述的通式(Ia)所示的化合物以及权利要求27所述的药物组合物在制备治疗ERK介导的相关疾病药物中的应用。The use of the compound represented by the general formula (Ia) according to any one of claims 1 to 7 and the pharmaceutical composition according to claim 27 in the preparation of drugs for treating ERK-mediated related diseases.
  27. 一种药物组合物，其含有治疗有效量的权利要求8-14任意一项所述的通式(I)所示的化合物的晶型，以及一种或多种药学上可接受的载体。A pharmaceutical composition containing a therapeutically effective amount of the crystal form of the compound represented by the general formula (I) according to any one of claims 8-14, and one or more pharmaceutically acceptable carriers.
  28. 根据权利要求权利要求8-14任意一项所述的通式(I)所示的化合物的晶型，以及权利要求27所述的药物组合物在制备治疗ERK介导的相关疾病药物中的应用。The crystal form of the compound represented by the general formula (I) according to any one of claims 8-14, and the use of the pharmaceutical composition of claim 27 in the preparation of drugs for treating ERK-mediated related diseases .
  29. 一种药物组合物，其含有治疗有效量的权利要求20～24任一项所述的式(IV)所示化合物的晶型，以及一种或多种药学上可接受的载体。A pharmaceutical composition comprising a therapeutically effective amount of the crystal form of the compound represented by formula (IV) according to any one of claims 20-24, and one or more pharmaceutically acceptable carriers.
  30. 根据权利要求20～24任一项所述的式(IV)所示化合物的晶型，以及权利要求31所述的药物组合物在制备治疗ERK介导的相关疾病药物中的应用。The crystal form of the compound represented by formula (IV) according to any one of claims 20-24, and the use of the pharmaceutical composition according to claim 31 in the preparation of drugs for treating ERK-mediated related diseases.
  31. 根据权利要求26、28或30所述的应用，其中所述的ERK介导的相关疾病选自癌症、骨病、炎性疾病、免疫疾病、神经系统疾病、代谢性疾病、呼吸性疾病或心脏病；其中所述癌症选自乳腺癌、胰腺癌、非小细胞肺癌、甲状腺癌、精原细胞瘤、黑素瘤、膀胱癌、肝癌、肾癌、骨髓增生异常综合征、急性髓性白血病或结直肠癌。The use according to claim 26, 28 or 30, wherein the ERK-mediated related disease is selected from cancer, bone disease, inflammatory disease, immune disease, neurological disease, metabolic disease, respiratory disease or heart disease Disease; wherein the cancer is selected from breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, acute myeloid leukemia or Colorectal cancer.
PCT/CN2020/082042 2019-04-02 2020-03-30 Salt of indazolyl-containing tricyclic derivative and crystal form thereof WO2020200161A1 (en)

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