WO2024234384A1 - 一种嘧啶并氮杂环类化合物的晶型及其制备方法及应用 - Google Patents

一种嘧啶并氮杂环类化合物的晶型及其制备方法及应用 Download PDF

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WO2024234384A1
WO2024234384A1 PCT/CN2023/095142 CN2023095142W WO2024234384A1 WO 2024234384 A1 WO2024234384 A1 WO 2024234384A1 CN 2023095142 W CN2023095142 W CN 2023095142W WO 2024234384 A1 WO2024234384 A1 WO 2024234384A1
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free base
crystal form
ray powder
powder diffraction
diffraction pattern
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French (fr)
Inventor
李钧
牛成山
郑茂林
梁阿朋
吴豫生
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TYK Medicines Inc
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TYK Medicines Inc
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Priority to EP23937064.6A priority Critical patent/EP4714947A4/en
Priority to JP2025567960A priority patent/JP2026518724A/ja
Priority to PCT/CN2023/095142 priority patent/WO2024234384A1/zh
Priority to CN202380097392.5A priority patent/CN121039115A/zh
Publication of WO2024234384A1 publication Critical patent/WO2024234384A1/zh
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    • 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
    • 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
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C57/00—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms
    • C07C57/02—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms with only carbon-to-carbon double bonds as unsaturation
    • C07C57/13—Dicarboxylic acids
    • C07C57/15—Fumaric acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C59/00—Compounds having carboxyl groups bound to acyclic carbon atoms and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
    • C07C59/235—Saturated compounds containing more than one carboxyl group
    • C07C59/245—Saturated compounds containing more than one carboxyl group containing hydroxy or O-metal groups
    • 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
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs

Definitions

  • the present invention relates to the field of medical technology, and in particular to a crystal form of a pyrimidine nitrogen heterocyclic compound, a preparation method and an application thereof.
  • the RET (Rearranged during transfection) gene is located on chromosome 10.
  • the RET protein it encodes is a receptor tyrosine kinase (RTK) present on the cell membrane.
  • RTK receptor tyrosine kinase
  • Its mutation types mainly include fusion mutations with genes such as KIF5B, TRIM33, CCDC6 and NCOA4, as well as point mutations at sites such as M918T.
  • Common RET mutations occur mainly in various cancer types such as thyroid cancer and non-small cell lung cancer.
  • the incidence of RET gene fusion in NSCLC patients is about 1% to 2%, and the incidence in papillary thyroid carcinoma (accounting for about 85% of all thyroid cancers) is 10% to 20%.
  • RET fusion is more common in young patients, especially young non-smoking lung adenocarcinoma patients, with an incidence of up to 7% to 17%.
  • the treatment plan for RET gene changes mainly uses multi-kinase inhibitor drugs, such as cabozantinib and vandetanib. Due to the low targeting, severe toxicity associated with VEGFR inhibition caused by off-target often occurs.
  • ORR overall response rate
  • the Chinese patent with application publication number CN113698390A discloses a series of compounds used as RET kinase inhibitors, and the series of compounds disclosed in this patent have very good RET inhibitory activity.
  • a compound with a structural formula as shown in formula (A) is disclosed. There is no report on the crystal form patent of the compound.
  • the object of the present invention is to provide a crystal form of a pyrimidoazaheterocyclic compound, and a preparation method and application thereof.
  • a crystalline form of a compound of formula A wherein the crystalline form is selected from the group consisting of free base crystalline form III, fumarate crystalline form I, and malate crystalline form I.
  • the X-ray powder diffraction pattern of the free base crystalline form III has characteristic peaks at the following 2 ⁇ values: 5.1 ⁇ 0.2°, 5.8 ⁇ 0.2°, 7.3 ⁇ 0.2°, 8.2 ⁇ 0.2°, 9.3 ⁇ 0.2°, 10.4 ⁇ 0.2°, 10.7 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.3 ⁇ 0.2°, 14.7 ⁇ 0.2°, 16.7 ⁇ 0.2°, 18.4 ⁇ 0.2°, 19.0 ⁇ 0.2°, 23.4 ⁇ 0.2°;
  • the X-ray powder diffraction spectrum of the fumarate salt form I has characteristic peaks at the following 2 ⁇ values: 6.0 ⁇ 0.2°, 8.7 ⁇ 0.2°, 10.6 ⁇ 0.2°, 14.6 ⁇ 0.2°, 15.5 ⁇ 0.2°, 17.5 ⁇ 0.2°, 18.0 ⁇ 0.2°, 19.2 ⁇ 0.2°, 27.1 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the malate crystalline form I has characteristic peaks at the following 2 ⁇ values: 6.0 ⁇ 0.2°, 9.3 ⁇ 0.2°, 10.7 ⁇ 0.2°, 11.2 ⁇ 0.2°, 11.8 ⁇ 0.2°, 15.3 ⁇ 0.2°, 17.3 ⁇ 0.2°, 17.6 ⁇ 0.2°, 18.3 ⁇ 0.2°, 19.0 ⁇ 0.2°, 23.5 ⁇ 0.2°, 25.7 ⁇ 0.2°, and 28.0 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the free base crystalline form III has characteristic peaks at the following 2 ⁇ values: 5.1 ⁇ 0.2°, 5.8 ⁇ 0.2°, 7.3 ⁇ 0.2°, 7.8 ⁇ 0.2°, 8.2 ⁇ 0.2°, 9.3 ⁇ 0.2°, 10.4 ⁇ 0.2°, 10.7 ⁇ 0.2°, 11.0 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.3 ⁇ 0.2°, 14.0 ⁇ 0.2°, 14.7 ⁇ 0.2°, 15.2 ⁇ 0.2°, 16.7 ⁇ 0.2°, 17.5 ⁇ 0.2°, 18.4 ⁇ 0.2°, 19.0 ⁇ 0.2°, 21.1 ⁇ 0.2°, 22.2 ⁇ 0.2°, 23.4 ⁇ 0.2°, 29.5 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the fumarate salt form I has characteristic peaks at the following 2 ⁇ values: 6.0 ⁇ 0.2°, 8.7 ⁇ 0.2°, 10.6 ⁇ 0.2°, 11.0 ⁇ 0.2°, 11.9 ⁇ 0.2°, 12.1 ⁇ 0.2°, 12.5 ⁇ 0.2°, 14.6 ⁇ 0.2°, 15.5 ⁇ 0.2°, 15.9 ⁇ 0.2°, 17.5 ⁇ 0.2°, 18.0 ⁇ 0.2°, 19.2 ⁇ 0.2°, 26.7 ⁇ 0.2°, 27.1 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the malate crystalline form I has characteristic peaks at the following 2 ⁇ values: 6.0 ⁇ 0.2°, 9.3 ⁇ 0.2°, 10.2 ⁇ 0.2°, 10.7 ⁇ 0.2°, 11.2 ⁇ 0.2°, 11.8 ⁇ 0.2°, 13.2 ⁇ 0.2°, 15.3 ⁇ 0.2°, 16.2 ⁇ 0.2°, 17.3 ⁇ 0.2°, 17.6 ⁇ 0.2°, 18.3 ⁇ 0.2°, 19.0 ⁇ 0.2°, 23.5 ⁇ 0.2°, 24.8 ⁇ 0.2°, 25.7 ⁇ 0.2°, 27.6 ⁇ 0.2°, and 28.0 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the free base crystalline form III has characteristic peaks at the following 2 ⁇ values: 5.1 ⁇ 0.2°, 5.8 ⁇ 0.2°, 7.3 ⁇ 0.2°, 7.8 ⁇ 0.2°, 8.2 ⁇ 0.2°, 9.3 ⁇ 0.2°, 10.4 ⁇ 0.2°, 10.7 ⁇ 0.2°, 11.0 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.3 ⁇ 0.2°, 14.0 ⁇ 0.2°, 14.7 ⁇ 0.2°, 15.2 ⁇ 0.2°.
  • the X-ray powder diffraction spectrum of the fumarate salt form I has characteristic peaks at the following 2 ⁇ values: 6.0 ⁇ 0.2°, 8.2 ⁇ 0.2°, 8.7 ⁇ 0.2°, 9.9 ⁇ 0.2°, 10.6 ⁇ 0.2°, 11.0 ⁇ 0.2°, 11.9 ⁇ 0.2°, 12.1 ⁇ 0.2°, 12.5 ⁇ 0.2°, 14.6 ⁇ 0.2°, 15.8 ⁇ 0.2°, 16.0 ⁇ 0.2°, 17.1 ⁇ 0.2°, 18.8 ⁇ 0.2°, 19.4 ⁇ 0.2°, 20.8 ⁇ 0.2°, 21.3 ⁇ 0.2°, 22.4 ⁇ 0.2°, 23.9 ⁇ 0.2°, 24.8 ⁇ 0.2°, 25.9 ⁇ 0.2°, 26.8 ⁇ 0.2°, 27.9 ⁇ 0.2°, 28.
  • the X-ray powder diffraction spectrum of the malate crystalline form I has characteristic peaks at the following 2 ⁇ values: 6.0 ⁇ 0.2°, 9.3 ⁇ 0.2°, 10.2 ⁇ 0.2°, 10.7 ⁇ 0.2°, 11.2 ⁇ 0.2°, 11.8 ⁇ 0.2°, 12.1 ⁇ 0.2°, 13.2 ⁇ 0.2°, 15.3 ⁇ 0.2°, 16.2 ⁇ 0.2°, 16.8 ⁇ 0.2°, 17.1 ⁇ 0.2°, 17.3 ⁇ 0.2°, 17.6 ⁇ 0.2°, 18.3 ⁇ 0.2°, 19.0 ⁇ 0.2°, 23.5 ⁇ 0.2°, 24.8 ⁇ 0.2°, 25.2 ⁇ 0.2°, 25.7 ⁇ 0.2°, 27.6 ⁇ 0.2°, 28.0 ⁇ 0.2°, 29.7 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the free base Form III is substantially as shown in FIG5 ;
  • the X-ray powder diffraction pattern of the fumarate salt form I is substantially as shown in FIG37 ;
  • the X-ray powder diffraction pattern of the malate crystalline form I is basically as shown in Figure 41.
  • the crystal form is selected from the following group: fumarate crystal form I and malate crystal form I.
  • the crystal form is fumarate crystal form I.
  • the fumarate salt crystal form I is triclinic system, P1 space group.
  • each asymmetric unit contains two free base molecules and one fumaric acid molecule, and each unit cell contains one asymmetric unit.
  • the second aspect of the present invention provides a method for preparing the free base crystal form III, wherein the method is selected from the following group:
  • Method 1 Mix the compound represented by formula A with solvent m, dissolve it, and crystallize it; the solvent m is selected from one of n-butyl ketone, acetone and acetonitrile;
  • Method 2 Mix the compound represented by formula A with solvent n, beat at high temperature, and crystallize out; the solvent n is selected from one of ethyl acetate, isopropyl acetate and methyl tert-butyl ether.
  • the compound represented by formula A is free base crystal form I.
  • the "high temperature” is 50-70°C.
  • the third aspect of the present invention provides a method for preparing fumarate crystalline form I or malate crystalline form I, wherein the method is selected from the following group:
  • Method 1 dissolving the compound of formula A in an organic solvent, stirring at room temperature, then adding fumaric acid or malic acid, continuing stirring, and performing solid-liquid separation to obtain the fumarate salt crystal form I or malate salt crystal form I;
  • Method 2 adding an organic solvent to the fumarate or malate of the compound of formula A, suspending and stirring, and performing solid-liquid separation to obtain the fumarate crystal form I or malate crystal form I.
  • the organic solvent is 2-methyltetrahydrofuran.
  • the fourth aspect of the present invention provides a use of the crystal form described in the first aspect of the present invention for preparing a drug, wherein the drug is used for a use selected from the group consisting of:
  • the disease or condition related to RET regulation is selected from the following group: thyroid cancer and non-small cell lung cancer.
  • the cells are selected from the group consisting of human thyroid cancer cells and human lung cancer cells.
  • the tumor is selected from the following group: thyroid cancer and non-small cell lung cancer.
  • FIG1 is an X-ray powder diffraction pattern of the free base crystalline form I of the present invention.
  • FIG2 is a DSC-TGA spectrum of the free base crystalline form I of the present invention.
  • FIG3 is an X-ray powder diffraction pattern of the free base crystalline form II of the present invention.
  • FIG4 is a DSC-TGA spectrum of the free base crystal form II of the present invention.
  • FIG5 is an X-ray powder diffraction pattern of the free base crystalline form III of the present invention.
  • FIG6 is a DSC-TGA spectrum of the free base form III of the present invention.
  • FIG7 is an X-ray powder diffraction pattern of the free base crystalline form IV of the present invention.
  • FIG8 is a DSC-TGA spectrum of the free base form IV of the present invention.
  • FIG9 is an X-ray powder diffraction pattern of the free base crystalline form V of the present invention.
  • FIG10 is a DSC-TGA spectrum of the free base crystalline form V of the present invention.
  • FIG11 is an X-ray powder diffraction pattern of the free base crystalline form VI of the present invention.
  • FIG12 is a DSC-TGA spectrum of the free base form VI of the present invention.
  • FIG13 is an X-ray powder diffraction pattern of the free base form VII of the present invention.
  • FIG14 is a DSC-TGA spectrum of the free base form VII of the present invention.
  • FIG15 is an X-ray powder diffraction pattern of the free base crystalline form VIII of the present invention.
  • FIG16 is a DSC-TGA spectrum of the free base Form VIII of the present invention.
  • FIG17 is an X-ray powder diffraction pattern of the free base Form IX of the present invention.
  • FIG18 is a DSC-TGA spectrum of the free base Form IX of the present invention.
  • FIG19 is an X-ray powder diffraction pattern of the hydrochloride salt form I of the present invention.
  • FIG20 is a DSC-TGA spectrum of hydrochloride crystal form I of the present invention.
  • FIG21 is an X-ray powder diffraction pattern of the p-toluenesulfonate crystalline form I of the present invention.
  • FIG22 is a DSC-TGA spectrum of p-toluenesulfonate crystalline form I of the present invention.
  • FIG23 is an X-ray powder diffraction pattern of the mesylate salt Form I of the present invention.
  • FIG24 is a DSC-TGA spectrum of the mesylate salt Form I of the present invention.
  • FIG25 is an X-ray powder diffraction pattern of the benzenesulfonate salt form I of the present invention.
  • FIG26 is a DSC-TGA spectrum of benzenesulfonate salt Form I of the present invention.
  • FIG28 is a DSC-TGA spectrum of oxalate crystal form I of the present invention.
  • FIG29 is an X-ray powder diffraction pattern of the oxalate crystal form II of the present invention.
  • FIG30 is a DSC-TGA spectrum of oxalate crystal form II of the present invention.
  • FIG31 is an X-ray powder diffraction pattern of maleate salt Form I of the present invention.
  • FIG32 is a DSC-TGA spectrum of maleate salt Form I of the present invention.
  • FIG33 is an X-ray powder diffraction pattern of the phosphate crystal form I of the present invention.
  • FIG34 is a DSC-TGA spectrum of phosphate crystal form I of the present invention.
  • FIG35 is an X-ray powder diffraction pattern of the tartrate salt form I of the present invention.
  • FIG36 is a DSC-TGA spectrum of the tartrate salt Form I of the present invention.
  • FIG37 is an X-ray powder diffraction pattern of the fumarate salt Form I of the present invention.
  • FIG39 is an X-ray powder diffraction pattern of the citrate crystalline form I of the present invention.
  • FIG40 is a DSC-TGA spectrum of the citrate crystalline form I of the present invention.
  • FIG41 is an X-ray powder diffraction pattern of the malate crystalline form I of the present invention.
  • FIG. 42 is a DSC-TGA spectrum of the malate crystal form I of the present invention.
  • Figure 43 is a single crystal structure diagram of the fumarate salt form I of the present invention (containing two free base molecules and one fumaric acid molecule, each unit cell contains an asymmetric unit, and the absolute configuration of carbon (C-8/C-36) is S configuration).
  • Figure 44 is an X-ray powder diffraction pattern of the amorphous free base of the present invention.
  • Figures 45 and 46 are PLM diagrams of the free base form I of the present invention.
  • the inventor unexpectedly prepared a series of crystalline forms of the compound represented by formula A with excellent stability, especially free base crystalline form III, fumarate crystalline form I, and malate crystalline form I. On this basis, the inventor completed the present invention.
  • Crystal form is the solid state of a drug.
  • the study of drug crystal form is the study of the basic state of the drug. Only when we have a relatively full and comprehensive understanding of the crystal state of chemical drugs can we find more suitable drug crystal solid substances for treating diseases.
  • the crystal form of a drug can affect the physical and chemical properties of the drug and directly affect the basis for the drug to play a role in treating diseases in clinical practice. Different crystal forms of the same drug may have significant differences in appearance, solubility, melting point, dissolution, bioavailability, etc., thereby affecting the stability, bioavailability and efficacy of the drug. Therefore, it is of great significance to study the stable crystal form of the compound.
  • the active ingredients of the same drug generally exist in two or more crystal forms, which are called drug polymorphs.
  • Different crystal forms have different solubility and dissolution rates, which affect the clinical therapeutic effect of the drug by causing changes in bioavailability in the body.
  • the difference in drug crystal forms may affect its dissolution and absorption in the body, and then affect the bioavailability, clinical efficacy and safety of the drug.
  • the stability of the drug crystal form is also very important. In order to improve the bioavailability of the drug, reduce toxicity, and enhance the therapeutic effect, more attention should be paid to the stability of the drug crystal form.
  • Crystal forms with good stability can ensure the physicochemical stability of the drug dosage form during preparation and storage, maintain good solubility and bioavailability of the drug dosage form, and ensure the equivalence between each batch of drugs.
  • the same drug often has multiple crystal forms. At present, the crystal form with better therapeutic effect and the most suitable clinically is called the dominant drug crystal form.
  • the present invention screened the crystal forms of the compound represented by formula (A) and found as many different crystal forms of the raw material drug as possible.
  • the screened crystal forms were identified by powder X-ray diffraction analysis (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), hydrogen spectrum liquid nuclear magnetic resonance (1H NMR) and high performance liquid chromatography (HPLC), and further determined the free base crystal form III, fumarate crystal form I and malate crystal form I of the dominant drug through thermodynamic stability and hygroscopicity research, providing a reference for the selection of crystal forms in subsequent drug metabolism dynamics experiments and animal experiments.
  • XRPD powder X-ray diffraction analysis
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • HPLC high performance liquid chromatography
  • the free base crystal form III, fumarate crystal form I and malate crystal form I shown in formula (A) provided by the present invention are easily absorbed and have good stability in vivo, have good blood drug concentration distribution in vivo, and have high bioavailability.
  • due to its good physical stability it is very beneficial to the quality control of the drug in the subsequent formulation development and has a longer shelf life, so that the drug can achieve better therapeutic effects.
  • the compound of formula A also has a crystalline form selected from the group consisting of free base crystalline form I, free base crystalline form II, free base crystalline form IV, free base crystalline form V, free base crystalline form VI, free base crystalline form VII, free base crystalline form VIII, free base crystalline form IX,
  • the X-ray powder diffraction pattern of the free base crystalline form I has characteristic peaks at the following 2 ⁇ values: 6.8 ⁇ 0.2°, 8.6 ⁇ 0.2°, 9.9 ⁇ 0.2°, 12.9 ⁇ 0.2°, 13.7 ⁇ 0.2°, 18.8 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form II has characteristic peaks at the following 2 ⁇ values: 5.4 ⁇ 0.2°, 8.1 ⁇ 0.2°, 9.7 ⁇ 0.2°, 10.0 ⁇ 0.2°, 10.5 ⁇ 0.2°, 13.6 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form IV has characteristic peaks at the following 2 ⁇ values: 5.7 ⁇ 0.2°, 8.3 ⁇ 0.2°, 9.5 ⁇ 0.2°, 11.8 ⁇ 0.2°, 12.2 ⁇ 0.2°, 12.5 ⁇ 0.2°, 12.9 ⁇ 0.2°, 13.4 ⁇ 0.2°, 14.4 ⁇ 0.2°, 15.4 ⁇ 0.2°, 17.3 ⁇ 0.2°, 18.4 ⁇ 0.2°, 18.6 ⁇ 0.2°, 20.2 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form V has characteristic peaks at the following 2 ⁇ values: 5.4 ⁇ 0.2°, 9.5 ⁇ 0.2°, 9.8 ⁇ 0.2°, 11.9 ⁇ 0.2°, 13.7 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form VI has characteristic peaks at the following 2 ⁇ values: 5.7 ⁇ 0.2°, 7.4 ⁇ 0.2°, 8.4 ⁇ 0.2°, 9.7 ⁇ 0.2°, 10.3 ⁇ 0.2°, 11.5 ⁇ 0.2°, 12.4 ⁇ 0.2°, 14.6 ⁇ 0.2°, 15.7 ⁇ 0.2°, 17.1 ⁇ 0.2°, 17.4 ⁇ 0.2°, 18.7 ⁇ 0.2°, 20.3 ⁇ 0.2°, 23.1 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form VII has characteristic peaks at the following 2 ⁇ values: 5.1 ⁇ 0.2°, 5.6 ⁇ 0.2°, 7.0 ⁇ 0.2°, 7.4 ⁇ 0.2°, 9.5 ⁇ 0.2°, 9.9 ⁇ 0.2°, 10.3 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.3 ⁇ 0.2°, 14.4 ⁇ 0.2°, 14.9 ⁇ 0.2°, 16.1 ⁇ 0.2°, 18.1 ⁇ 0.2°, 18.6 ⁇ 0.2°, 18.9 ⁇ 0.2°, 20.6 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base form VIII has characteristic peaks at the following 2 ⁇ values: 5 ⁇ 0.2°, 8.3 ⁇ 0.2°, 8.6 ⁇ 0.2°, 10.1 ⁇ 0.2°, 11.2 ⁇ 0.2°, 12.4 ⁇ 0.2°, 12.6 ⁇ 0.2°, 13.1 ⁇ 0.2°, 13.6 ⁇ 0.2°, 17.7 ⁇ 0.2°, 18 ⁇ 0.2°, 19.2 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base form IX has characteristic peaks at the following 2 ⁇ values: 7.9 ⁇ 0.2°, 11.8 ⁇ 0.2°, 15.9 ⁇ 0.2°, 16.4 ⁇ 0.2°, 17.3 ⁇ 0.2°, 20.0 ⁇ 0.2°, 20.2 ⁇ 0.2°, and 21.2 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the free base crystalline form I has characteristic peaks at the following 2 ⁇ values: 5.8 ⁇ 0.2°, 6.8 ⁇ 0.2°, 8.6 ⁇ 0.2°, 9.9 ⁇ 0.2°, 12.0 ⁇ 0.2°, 12.9 ⁇ 0.2°, 13.7 ⁇ 0.2°, 14.4 ⁇ 0.2°, 15.2 ⁇ 0.2°, 16.5 ⁇ 0.2°, 17.2 ⁇ 0.2°, 17.6 ⁇ 0.2°, 18.4 ⁇ 0.2°, 18.8 ⁇ 0.2°, 21.9 ⁇ 0.2°, 23.0 ⁇ 0.2°, 23.5 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form II has characteristic peaks at the following 2 ⁇ values: 5.4 ⁇ 0.2°, 8.1 ⁇ 0.2°, 9.7 ⁇ 0.2°, 10.0 ⁇ 0.2°, 10.5 ⁇ 0.2°, 11.8 ⁇ 0.2°, 12.6 ⁇ 0.2°, 13.6 ⁇ 0.2°, 16.4 ⁇ 0.2°, 19.8 ⁇ 0.2°, 23.0 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form IV has characteristic peaks at the following 2 ⁇ values: 5.7 ⁇ 0.2°, 6.6 ⁇ 0.2°, 6.7 ⁇ 0.2°, 7.2 ⁇ 0.2°, 8.3 ⁇ 0.2°, 9.5 ⁇ 0.2°, 10.0 ⁇ 0.2°, 11.5 ⁇ 0.2°, 11.8 ⁇ 0.2°, 12.2 ⁇ 0.2°, 12.5 ⁇ 0.2°, 12.9 ⁇ 0.2°, 13.1 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.1 ⁇ 0.2°, 13.2 ⁇ 0.2°, 13.3 ⁇ 0.2°, 13.4 ⁇ 0.2°, 13.6 ⁇ 0.2°, 13.8 ⁇ 0.2°, 13.9 ⁇ 0.2°, 13.1 ⁇ 0.2°, 13.9 ⁇ 0.2°, 13.1 ⁇ 0.2°, 13.3 ...
  • the X-ray powder diffraction pattern of the free base crystalline form V has characteristic peaks at the following 2 ⁇ values: 5.4 ⁇ 0.2°, 8.2 ⁇ 0.2°, 9.5 ⁇ 0.2°, 9.8 ⁇ 0.2°, 11.1 ⁇ 0.2°, 11.9 ⁇ 0.2°, 12.8 ⁇ 0.2°, 13.7 ⁇ 0.2°, 16.4 ⁇ 0.2°, 19.2 ⁇ 0.2°, 20.9 ⁇ 0.2°, 23.2 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form VI has characteristic peaks at the following 2 ⁇ values: 5.7 ⁇ 0.2°, 6.7 ⁇ 0.2°, 7.4 ⁇ 0.2°, 8.4 ⁇ 0.2°, 9.7 ⁇ 0.2°, 10.3 ⁇ 0.2°, 11.5 ⁇ 0.2°, 12.0 ⁇ 0.2°, 12.4 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.6 ⁇ 0.2°, 14.6 ⁇ 0.2°, 15.7 ⁇ 0.2°, 16.2 ⁇ 0.2°, 17.1 ⁇ 0.2°, 17.4 ⁇ 0.2°, 18.7 ⁇ 0.2°, 20.3 ⁇ 0.2°, 21.4 ⁇ 0.2°, 22.8 ⁇ 0.2°, 23.1 ⁇ 0.2°, 29.9 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form VII has characteristic peaks at the following 2 ⁇ values: 5.1 ⁇ 0.2°, 5.6 ⁇ 0.2°, 7.0 ⁇ 0.2°, 7.4 ⁇ 0.2°, 7.8 ⁇ 0.2°, 8.7 ⁇ 0.2°, 9.5 ⁇ 0.2°, 9.9 ⁇ 0.2°, 10.3 ⁇ 0.2°, 11.7 ⁇ 0.2°, 12 .1 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.3 ⁇ 0.2°, 13.8 ⁇ 0.2°, 14.4 ⁇ 0.2°, 14.9 ⁇ 0.2°, 16.1 ⁇ 0 .2°, 16.3 ⁇ 0.2°, 18.1 ⁇ 0.2°, 18.6 ⁇ 0.2°, 18.9 ⁇ 0.2°, 20.6 ⁇ 0.2°, 23.5 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base form VIII has characteristic peaks at the following 2 ⁇ values: 4.3 ⁇ 0.2°, 5 ⁇ 0.2°, 5.5 ⁇ 0.2°, 8.3 ⁇ 0.2°, 8.6 ⁇ 0.2°, 10.1 ⁇ 0.2°, 10.4 ⁇ 0.2°, 11.2 ⁇ 0.2°, 12.4 ⁇ 0.2°, 12.6 ⁇ 0.2°, 13.1 ⁇ 0.2°, 13.6 ⁇ 0.2°, 14.4 ⁇ 0.2°, 16.6 ⁇ 0.2°, 17.7 ⁇ 0.2°, 18 ⁇ 0.2°, 19.2 ⁇ 0.2°, 20 .3 ⁇ 0.2°, 22 ⁇ 0.2°, 23.6 ⁇ 0.2°, 24.9 ⁇ 0.2°, 25.1 ⁇ 0.2°, 26.1 ⁇ 0.2°, 26.4 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base form IX has characteristic peaks at the following 2 ⁇ values: 7.9 ⁇ 0.2°, 8.4 ⁇ 0.2°, 11.8 ⁇ 0.2°, 13.8 ⁇ 0.2°, 15.9 ⁇ 0.2°, 16.4 ⁇ 0.2°, 17.3 ⁇ 0.2°, 19.0 ⁇ 0.2°, 19.2 ⁇ 0.2°, 20.0 ⁇ 0.2°, 20.2 ⁇ 0.2°, 21.2 ⁇ 0.2°, and 28.4 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the free base crystalline form I has characteristic peaks at the following 2 ⁇ values: 5.8 ⁇ 0.2°, 6.8 ⁇ 0.2°, 8.6 ⁇ 0.2°, 9.9 ⁇ 0.2°, 12.0 ⁇ 0.2°, 12.9 ⁇ 0.2°, 13.7 ⁇ 0.2°, 14.4 ⁇ 0.2°, 15.2 ⁇ 0.2°, 16.5 ⁇ 0.2°, 17.2 ⁇ 0.2°, 17.6 ⁇ 0. 2°, 18.1 ⁇ 0.2°, 18.4 ⁇ 0.2°, 18.8 ⁇ 0.2°, 19.5 ⁇ 0.2°, 20.5 ⁇ 0.2°, 21.6 ⁇ 0.2°, 21.9 ⁇ 0.2°, 22. 3 ⁇ 0.2°, 23.0 ⁇ 0.2°, 23.5 ⁇ 0.2°, 25.1 ⁇ 0.2°, 25.7 ⁇ 0.2°, 26.1 ⁇ 0.2°, 29.6 ⁇ 0.2°, 29.9 ⁇ 0.2°;
  • the X-ray powder diffraction spectrum of the free base crystalline form II has characteristic peaks at the following 2 ⁇ values: 5.4 ⁇ 0.2°, 6.6 ⁇ 0.2°, 8.1 ⁇ 0.2°, 9.1 ⁇ 0.2°, 9.7 ⁇ 0.2°, 10.0 ⁇ 0.2°, 10.5 ⁇ 0.2°, 11.4 ⁇ 0.2°, 11.8 ⁇ 0.2°, 12.6 ⁇ 0.2°, 13.9 ⁇ 0.2°, 14.8 ⁇ 0.2°, 15.3 ⁇ 0.2°, 16.7 ⁇ 0.2°, 17.9 ⁇ 0.2°, 18.4 ⁇ 0.2°, 19.
  • the X-ray powder diffraction pattern of the free base crystalline form IV has characteristic peaks at the following 2 ⁇ values: 3.5 ⁇ 0.2°, 4.1 ⁇ 0.2°, 5.3 ⁇ 0.2°, 5.7 ⁇ 0.2°, 6.6 ⁇ 0.2°, 6.7 ⁇ 0.2°, 7.2 ⁇ 0.2°, 8.3 ⁇ 0.2°, 9.5 ⁇ 0.2°, 10.0 ⁇ 0.2°, 10.2 ⁇ 0.2°, 10.7 ⁇ 0.2°, 11.5 ⁇ 0.2°, 11.8 ⁇ 0.2°, 12.2 ⁇ 0.2°, 12.5 ⁇ 0.2°, 12.9 ⁇ 0.2° , 13.1 ⁇ 0.2°, 13.4 ⁇ 0.2°, 14.4 ⁇ 0.2°, 14.6 ⁇ 0.2°, 15.4 ⁇ 0.2°, 15.6 ⁇ 0.2°, 16.
  • the X-ray powder diffraction pattern of the free base crystalline form V has characteristic peaks at the following 2 ⁇ values: 5.4 ⁇ 0.2°, 8.2 ⁇ 0.2°, 9.5 ⁇ 0.2°, 9.8 ⁇ 0.2°, 11.1 ⁇ 0.2°, 11.9 ⁇ 0.2°, 12.8 ⁇ 0.2°, 13.7 ⁇ 0.2°, 14.9 ⁇ 0.2°, 16.4 ⁇ 0.2°, 17.1 ⁇ 0.2°, 18.4 ⁇ 0.2°, 19.8 ⁇ 0.2°, 20.3 ⁇ 0.2°, 21.1 ⁇ 0.2°, 22.8 ⁇ 0.2°, 23.8 ⁇ 0.2°, 24.8 ⁇ 0.2°, 25.8 ⁇ 0.2°, 26.6 ⁇ 0.2°, 27.8 ⁇ 0.2°, 28.8 ⁇ 0.2°, 29.
  • the X-ray powder diffraction spectrum of the free base crystalline form VI has characteristic peaks at the following 2 ⁇ values: 4.2 ⁇ 0.2°, 5.7 ⁇ 0.2°, 6.7 ⁇ 0.2°, 7.4 ⁇ 0.2°, 8.4 ⁇ 0.2°, 9.7 ⁇ 0.2°, 10.3 ⁇ 0.2°, 11.5 ⁇ 0.2°, 12.0 ⁇ 0.2°, 12.4 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.6 ⁇ 0.2°, 14.6 ⁇ 0.2°, 15 .7 ⁇ 0.2°, 16.2 ⁇ 0.2°, 17.1 ⁇ 0.2°, 17.4 ⁇ 0.2°, 18.7 ⁇ 0.2°, 20.3 ⁇ 0.2°, 21.0 ⁇ 0.2°, 21.4 ⁇ 0.2°, 2 2.8 ⁇ 0.2°, 23.1 ⁇ 0.2°, 25.8 ⁇ 0.2°, 26.3 ⁇ 0.2°, 27.2 ⁇ 0.2°, 28.8 ⁇ 0.2°, 29.2 ⁇ 0.2°, 29.9 ⁇ 0.2°;
  • the X-ray powder diffraction pattern of the free base crystalline form VII has characteristic peaks at the following 2 ⁇ values: 5.1 ⁇ 0.2°, 5.6 ⁇ 0.2°, 7.0 ⁇ 0.2°, 7.4 ⁇ 0.2°, 7.8 ⁇ 0.2°, 8.7 ⁇ 0.2°, 9.5 ⁇ 0.2°, 9.9 ⁇ 0.2°, 10.3 ⁇ 0.2°, 11.4 ⁇ 0.2°, 11.7 ⁇ 0.2°, 12.1 ⁇ 0.2°, 13.0 ⁇ 0.2°, 13.3 ⁇ 0.2°, 13.8 ⁇ 0.2°, 14.
  • the X-ray powder diffraction pattern of the free base form VIII has characteristic peaks at the following 2 ⁇ values: 4.3 ⁇ 0.2°, 5 ⁇ 0.2°, 5.5 ⁇ 0.2°, 6.7 ⁇ 0.2°, 8.3 ⁇ 0.2°, 8.6 ⁇ 0.2°, 9.7 ⁇ 0.2°, 10.1 ⁇ 0.2°, 10.4 ⁇ 0.2°, 11.2 ⁇ 0.2°, 12.4 ⁇ 0.2°, 12.6 ⁇ 0.2°, 13.1 ⁇ 0.2°, 13.6 ⁇ 0.2°, 14.4 ⁇ 0.2°, 15 ⁇ 0.2°, 15.6 ⁇ 0.2°, 16.6 ⁇ 0.2° , 17.7 ⁇ 0.2°, 18 ⁇ 0.2°, 18.6 ⁇ 0.2°, 19.2 ⁇ 0.2°, 20.3 ⁇ 0.2°, 21 ⁇ 0.2°, 21.3 ⁇ 0.2°, 22 ⁇ 0.2°, 22.6 ⁇ 0.2°, 23.1 ⁇ 0.2°, 23.6 ⁇ 0 .2°, 24.9 ⁇ 0.2°, 25.1 ⁇ 0.2°, 25.7 ⁇ 0.2°, 26.1 ⁇ 0.2°, 26.4 ⁇ 0.2°, 27 ⁇ 0.2°, 27.5 ⁇ 0.2°, 28.9 ⁇ 0.2°, 30.3 ⁇ 0.2°
  • the X-ray powder diffraction pattern of the free base crystalline form IX has characteristic peaks at the following 2 ⁇ values: 5.5 ⁇ 0.2°, 5.8 ⁇ 0.2°, 6.8 ⁇ 0.2°, 7.9 ⁇ 0.2°, 8.4 ⁇ 0.2°, 8.6 ⁇ 0.2°, 9.8 ⁇ 0.2°, 11.8 ⁇ 0.2°, 13.3 ⁇ 0.2°, 13.8 ⁇ 0.2°, 14.0 ⁇ 0.2°, 15 .9 ⁇ 0.2°, 16.4 ⁇ 0.2°, 17.3 ⁇ 0.2°, 17.6 ⁇ 0.2°, 19.0 ⁇ 0.2°, 19.2 ⁇ 0.2°, 20.0 ⁇ 0.2°, 2 0.2 ⁇ 0.2°, 20.6 ⁇ 0.2°, 21.2 ⁇ 0.2°, 24.2 ⁇ 0.2°, 27.8 ⁇ 0.2°, 28.4 ⁇ 0.2°, 29.4 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of the free base crystalline form I is substantially as shown in FIG1 ;
  • the X-ray powder diffraction pattern of the free base crystal form II is substantially as shown in FIG3 ;
  • the X-ray powder diffraction pattern of the free base Form IV is substantially as shown in FIG7 ;
  • the X-ray powder diffraction pattern of the free base Form V is substantially as shown in FIG9 ;
  • the X-ray powder diffraction pattern of the free base Form VI is substantially as shown in FIG11 ;
  • the X-ray powder diffraction pattern of the free base Form VII is substantially as shown in FIG13 ;
  • the X-ray powder diffraction pattern of the free base Form VIII is substantially as shown in FIG15 ;
  • the X-ray powder diffraction pattern of the free base Form IX is substantially as shown in FIG. 17 .
  • the present invention has the following main advantages:
  • the crystal form has excellent stability and high solubility
  • reaction solution was diluted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and separated and purified by high performance liquid chromatography to obtain formula (A), Ms[M+H]589 and formula (B), MS[M+H]589.
  • the obtained solid of formula (A) was characterized by XRPD. The result is shown in FIG1 , and the obtained solid is free base crystalline form I.
  • the free base crystal form I is used as a raw material for the following crystal preparation and salt preparation experiments.
  • the solid crystal form screening of the compound of formula (A) is to obtain metastable and stable crystal forms by crystallization or precipitation from a selected single solvent and a mixed solvent thereof.
  • the methods used include evaporation crystallization, slurry crystallization, cooling crystallization, and anti-solvent precipitation.
  • PLM analysis was performed using an ECLIPSE LV100POL (Nikon, JPN) polarizing microscope. A small amount of sample was spread on a glass slide, cedar oil was dropped to disperse the sample, and a cover glass was added. Subsequently, the sample was placed under a microscope and observed using a 4-20x objective lens.
  • the solid sample was tested using an X-ray diffractometer.
  • the sample was spread flat on a zero-background single crystal silicon sample plate, gently pressed and flattened, and then analyzed according to the parameters in Table 2.
  • the TGA data of the samples were collected using a TA instrument. 1-5 mg of the sample was placed in a peeled open aluminum sample pan, the sample was heated according to the instrument parameters in Table 3, and the data was analyzed and processed using TRIOS software.
  • the DSC data of the samples were collected using TA Instruments. 1-3 mg of the sample was placed in a DSC sample pan with holes, and the heating measurement was performed according to the parameters in Table 4. The data was analyzed and processed using TRIOS software.
  • the free base crystal form I was obtained by slurrying at room temperature and 50°C in the MeOH, EtOH and IPA system, and by anti-solvent precipitation in the IPA/THF system.
  • the free base crystal form I is dissolved in trifluoroacetic acid, acetonitrile and water system, and the obtained solution is adjusted to alkalinity with sodium bicarbonate. After the organic solvent is concentrated under reduced pressure, the suspension is filtered, and the solid is washed with water to obtain the free base crystal form II.
  • the free base crystal form III was obtained by slurrying at room temperature or high temperature of 50/70°C for n-butyl ketone, acetone and acetonitrile, and at high temperature of 50/70°C for ethyl acetate, isopropyl acetate and methyl tert-butyl ether.
  • the free alkali crystal form III is agglomerated micro-crystals with high crystallinity.
  • the XRPD results of the free alkali crystal form III are shown in Figure 5, and the DSC and TGA results are shown in Figure 6.
  • TGA shows that there is no obvious weight loss before 150°C.
  • 1H-NMR shows that the sample contains 0.03% acetone residue.
  • There is a small endothermic peak at 84°C on the DSC curve (the crystal form does not change when heated to 100°C).
  • Form IV was prepared by fast/slow evaporation crystallization in 2-Me-THF or fast evaporation of THF.
  • the free base crystal form IV is agglomerated small crystals with low crystallinity.
  • the XRPD results of the free base crystal form IV are shown in Figure 7, and the DSC and TGA results are shown in Figure 8.
  • TGA shows a 36.9% weight loss before 210°C, which is attributed to desolvation.
  • DSC shows multiple endothermic peaks before 250°C. It becomes amorphous after heating to 170°C. Therefore, crystal form IV is a hydrate or a solvate.
  • the free base crystal form V is obtained by slow volatilization of MEK or DCM system, slurrying of DCM or cooling crystallization.
  • the samples obtained from the MEK system were characterized.
  • the free base form V is a needle-shaped product with medium crystallinity. Crystal.
  • the XRPD results of the free base crystal form V are shown in Figure 9, and the DSC and TGA results are shown in Figure 10.
  • the TGA curve shows a 5.2% weight loss at 60-170°C, which is attributed to desolvation.
  • DSC shows multiple endothermic peaks before 250°C, and the free base crystal form V is converted to an amorphous form after heating to 175°C. Therefore, the free base crystal form V is a hydrate or a solvate.
  • Free base form VI was prepared from the slow volatilization of THF and characterized. Free base form VI is agglomerated small crystals with medium crystallinity. The DSC and TGA results are shown in Figure 12. TGA shows a 2.6% weight loss before 90°C and a 5.5% weight loss from 90-195°C, which is attributed to the removal of the solvent. Free base form VI is converted into an amorphous form after being heated to 195°C. DSC detected multiple endothermic peaks before 250°C, so free base form VI may be a solvate or a hydrate.
  • Free base crystal form VII is obtained by slurrying in a 2-Me-THF system at room temperature or at a high temperature of 50°C for 7 days. Free base crystal form VII is agglomerated small particle crystals with medium crystallinity. 1H-NMR detected that the sample contained 0.97% 2-Me-THF residue. DSC and TGA results are shown in Figure 14. TGA showed a 1.7% weight loss between 85-240°C, which was attributed to the removal of the solvent. DSC detected two overlapping endothermic peaks at 230°C, which were attributed to the melting point. The crystal form of free base crystal form VII did not change after being heated to 210°C. Therefore, free base crystal form VII is anhydrous crystalline form.
  • the free base form VIII was obtained by slurrying in 50°C water for 7 days.
  • the XRPD spectra of the wet and dry samples are not completely consistent, and the peaks of the free base form VIII shift to a certain extent in different states.
  • the DSC and TGA results are shown in Figure 16. After the sample was dried at 40°C, the TGA results showed that there was still more than 3.2% weight loss before 160°C. DSC showed multiple endothermic peaks before 250°C. 1H-NMR did not detect obvious solvent residues.
  • the compound was heated to 180°C and the crystal form was converted into an amorphous form. Therefore, the free base form VIII may be a hydrate.
  • the free base Form IX was obtained from the antisolvent precipitation of DMSO/H 2 O.
  • the free base Form IX is a moderately crystalline
  • TGA shows a 2.1% weight loss before 85°C and an 11.5% weight loss at 85-260°C, which is attributed to the removal of the solvent.
  • DSC shows a broad endothermic peak at 37°C and another endothermic peak at 140°C. Therefore, the free base form IX may be a DMSO solvent compound.
  • the free base Form I sample was weighed and dissolved in 20 mL of dichloromethane at room temperature. After filtering, the filtrate was quickly freed from the solvent by rotary evaporation. The obtained solid sample was subjected to corresponding characterization tests, and its XRD spectrum is shown in FIG44 .
  • the compound of the structure shown in formula (A) showed a relatively complex polymorph phenomenon in the polymorph screening, and a total of 9 crystalline forms were obtained, namely, crystalline forms I-IX. Two of them were anhydrous crystalline forms (crystalline form III and crystalline form VII), and crystalline form III was relatively stable, with a higher melting point and enthalpy value, while the others were solvates or hydrates, and the data were summarized in Table 14.
  • the 2 ⁇ angle value of the X-ray powder diffraction peak has an error range of ⁇ 0.2 °. It should be understood that the 2 ⁇ value of the X-ray powder diffraction pattern may vary slightly between machines and samples, and its numerical range may differ within ⁇ 0.2 units, so the quoted values cannot be interpreted as absolute values.
  • the free base crystal form III is an anhydrous crystalline form with a high melting point (245° C.).
  • the free base crystal form III samples obtained by different solvents or methods are all small-particle crystals.
  • the free base crystal form III provided by the present invention has stable physical and chemical properties and is very suitable for application in subsequent drug research and development such as preparations.
  • the crystal forms I to IX provided by the present invention can be used to prepare RET inhibitor drugs, especially the crystal form III.
  • the 12 acids used in the salt type screening of the present invention are shown in Table 15.
  • free base i.e., free base crystal form I prepared above
  • an acid solution or solid acid 1.1 eq. or 0.6 eq.
  • different crystallization methods were tried to prepare the crystalline salt, such as cooling crystallization, volatile crystallization, and anti-solvent precipitation.
  • the solid was collected by filtration, vacuum dried, and further analyzed.
  • Hydrochloride Form I is agglomerated fine crystals with medium crystallinity. 1H-NMR shows that the sample has no obvious organic solvent residue. DSC and TGA results are shown in Figure 20. TGA shows that there is a 1.7% weight loss before 150°C, which is the removal of water. There is a broad endothermic peak in the initial stage of DSC heating, and two other endothermic peaks are detected at 173°C and 218°C. Therefore, hydrochloride Form I may be a hydrate.
  • the p-toluenesulfonate salt form I has agglomerated fine crystals with high crystallinity.
  • 1H-NMR detected 3.0% acetone residue, and the molar ratio of acid and base was 1:1.
  • the DSC and TGA results are shown in Figure 22.
  • the TGA results showed that there was a 3.0% weight loss before 210°C, which was the removal of acetone.
  • the DSC results showed a broad endothermic peak at 163°C and another endothermic peak at 212°C. Therefore, the p-toluenesulfonate salt form I may be a solvent compound.
  • the mesylate salt form I is agglomerated fine crystals with medium crystallinity. 1H-NMR shows no obvious organic solvent residue, and the acid-base molar ratio is 1:1.
  • the DSC and TGA results are shown in Figure 24.
  • the TGA results show that there is a 0.4% weight loss before 100°C, which is attributed to the removal of water.
  • the DSC results show a small endothermic peak at 154°C and another endothermic peak at 192°C, which is caused by the melting of the sample. Therefore, the mesylate salt form I may be an anhydrous form.
  • Benzenesulfonate salt form I is agglomerated fine crystals with high crystallinity. 1H-NMR shows that there is 0.19% acetone residue and the acid-base molar ratio is 1:1. The DSC and TGA results are shown in Figure 26. The TGA results show that there is no obvious weight loss before 200°C. The DSC results show that there is an endothermic peak at 235°C, which is the melting of the compound. Therefore, benzenesulfonate salt form I is an anhydrous crystalline form.
  • Oxalate Form I is a small particle crystal with high crystallinity. 1H-NMR shows no obvious methanol residue. DSC and TGA results are shown in Figure 28. TGA results show a 0.2% weight loss between 55-85°C, which is attributed to the removal of water. DSC shows two small endothermic peaks at 43°C and 81°C, and another endothermic peak at 229°C, which is attributed to the removal and melting of water. Therefore, oxalate Form I may be an anhydrous form.
  • Oxalate Form II is agglomerated fine crystals with high crystallinity.
  • 1H-NMR shows 0.07% acetone residue.
  • DSC and TGA results are shown in Figure 30. TGA shows no obvious weight loss before 200°C.
  • DSC shows an endothermic peak at 227°C, which is caused by sample melting. Therefore, oxalate Form II is anhydrous crystalline form.
  • Maleate salt form I has small crystals with high crystallinity. 1H-NMR shows that the sample contains 0.41% 2-Me-THF residue, and the molar ratio of acid to base is 1:1. DSC and TGA results are shown in Figure 32. TGA results show that there is no obvious weight loss before 150°C. DSC shows an endothermic peak at 175°C, which is the melting point of the compound. Therefore, maleate salt form I is an anhydrous crystalline form.
  • Phosphate crystal form I is a tiny crystal with high crystallinity. 1H-NMR shows about 0.06% acetone residue. DSC and TGA results are shown in Figure 34. TGA results show no obvious weight loss before 200°C. DSC shows an endothermic peak at 221°C, which is attributed to the melting point of the compound. Therefore, phosphate crystal form I may be an anhydrous crystal form.
  • Tartrate Form I is agglomerated fine crystals with medium crystallinity. 1H-NMR shows about 0.08% acetone residue and the acid-base molar ratio is 1:1. DSC and TGA results are shown in Figure 36. TGA results show 1.7% weight loss before 105°C and 0.6% weight loss between 105-165°C. DSC shows a broad endothermic peak at 40°C and two other endothermic peaks at 146°C and 181°C, which are attributed to dehydration. Therefore, Tartrate Form I may be a hydrate.
  • Fumarate salt form I has agglomerated fine crystals with high crystallinity.
  • 1H-NMR shows that the acid-base molar ratio is 1:2.
  • DSC and TGA results are shown in Figure 38. TGA results show that there is no obvious weight loss before 200°C.
  • DSC shows an endothermic peak at 239°C, which is attributed to the melting point of the compound. Therefore, fumarate salt form I is an anhydrous crystalline form.
  • Citrate Form I has agglomerated fine crystals with medium crystallinity.
  • 1H-NMR shows about 0.08% acetone residue and the acid-base molar ratio is 1:2.
  • DSC and TGA results are shown in Figure 40. TGA results show a 4.1% weight loss between 120-230°C, which is the removal of water.
  • DSC shows two broad endothermic peaks at 85°C and 155°C, and another endothermic peak at 230°C, which is attributed to dehydration and melting point. Therefore, citrate Form I may be a hydrate.
  • Malate Form I has agglomerated fine crystals with high crystallinity. 1H-NMR shows that it contains 0.06% acetone residue and the acid-base molar ratio is 1:2. The DSC and TGA results are shown in Figure 42. The TGA results show that there is a 0.2% weight loss before 60°C, which is attributed to the removal of water and acetone. DSC shows an endothermic peak at 217°C, which is attributed to the melting point. Therefore, malate Form I is an anhydrous crystalline form.
  • PLM analysis was performed using an ECLIPSE LV100POL (Nikon, JPN) polarizing microscope. A small amount of sample was spread on a glass slide, cedar oil was dropped to disperse the sample, and a cover glass was added. Subsequently, the sample was placed under a microscope and observed using a 4-20x objective lens.
  • the solid sample was tested using an X-ray diffractometer.
  • the sample was spread flat on a zero-background single crystal silicon sample plate, gently pressed and flattened, and then analyzed according to the parameters in Table 32.
  • the TGA data of the samples were collected using TA instruments. 1-5 mg of the sample was placed in a peeled open aluminum sample pan, the sample was heated according to the instrument parameters in Table 33, and the data was analyzed and processed using TRIOS software.
  • the DSC data of the samples were collected using TA Instruments. 1-3 mg of the sample was placed in a DSC sample pan with holes, and the heating was measured according to the parameters in Table 34. The data was analyzed and processed using TRIOS software.
  • the moisture adsorption and desorption analysis of the samples was performed using a DVS instrument. A certain amount of sample was placed in a peeled sample pan, the sample weight was automatically weighed, and the sample was analyzed according to the parameters in Table 35.
  • HPLC analysis was performed using an Agilent HPLC 1260 series instrument.
  • the HPLC method for solubility and stability testing is shown in Table 37.
  • fumarate crystal form I, malate crystal form I and free base crystal form III were selected for solubility, stability studies and drug metabolism studies.
  • the solubility of the salts was calculated using the free base.
  • the crystal stability of fumarate crystal form I, malate crystal form I and free base crystal form III is good, and the crystal forms of the three compounds under the two test conditions are unchanged.
  • the chemical purity of fumarate crystal form I is slightly reduced under 40°C/75%RH conditions, but the stability is also good under closed high temperature conditions, and the humidity needs to be controlled during storage.
  • the other two crystal forms have good chemical stability under both conditions.
  • the free base amorphous sample is unstable and can be easily converted into other crystalline forms in the solvent.
  • Fumarate Form I, malate Form I, free base Form I and free base Form III were subjected to in vivo drug metabolism experiments in rats and dogs.
  • SD rats were given a single oral administration of fumarate crystal form I, malate crystal form I, free base crystal form I and free base crystal form III, and blood samples were collected at different time points to determine the drug concentration in the rat plasma after administration of the test substances and calculate the relevant pharmacokinetic parameters.
  • Fumarate crystal form I, free base crystal form I and free base crystal form III were administered orally to dogs once, and blood samples were collected at different time points to determine the drug concentration in the dog's plasma after administration of the test substances and calculate the relevant pharmacokinetic parameters.
  • the preparation method is as follows:
  • the solvents for fumarate form I, malate form I, free base form I and free base form III are 0.5% MC + 1% tween 80.
  • the specific preparation method is: take 10 mg of the test sample, add 0.5% MC and 1% tween 80 and vortex mix to obtain a solution with a concentration of 1 mg/mL; prepare freshly before use, vortex mix, and set aside.
  • the prepared test solution was tested and analyzed by the analytical department of this laboratory using LC-MS/MS.
  • the blood sampling time points are:
  • Oral administration 0.25, 0.5, 1, 2, 4, 6, 8, 12 and 24h after administration. ( ⁇ 2min for time points ⁇ 2hr, ⁇ 15min for other time points.)
  • Plasma samples were analyzed by LC-MS/MS at the laboratory analytical department.
  • the pharmacokinetic parameters AUC 0 ⁇ t , AUC 0 ⁇ , C max , T max and T 1/2 of the test article and their mean values and standard deviations were calculated using the pharmacokinetic calculation software WinNonlin non-compartmental model.
  • the fumarate form I and malate form I have excellent in vivo exposure in animals, good solid-state stability, and are suitable for scale-up production and further research and development.

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Abstract

本发明涉及一种嘧啶并氮杂环类化合物的晶型及其制备方法及应用。具体地，本发明公开了一系列式A化合物的晶型，所述晶型具有优异的稳定性。本发明还公开了上述晶型的制法及其在预防和/或治疗肿瘤方面的应用。

Description

一种嘧啶并氮杂环类化合物的晶型及其制备方法及应用 技术领域
本发明涉及医药技术领域，具体地涉及一种嘧啶并氮杂环类化合物的晶型及其制备方法及应用。
背景技术
RET(Rearranged during transfection，转染期间重排)基因位于10号染色体，它所编码的RET蛋白是一种存在于细胞膜上的受体酪氨酸激酶(RTK)，其变异类型主要包括与KIF5B、TRIM33、CCDC6和NCOA4等基因的融合突变，以及M918T等位点的点突变。人们常见的RET突变主要发生在甲状腺癌、非小细胞肺癌等多种癌症类型里。RET基因融合在NSCLC患者中的发生率约为1％～2％，在甲状腺乳头状癌(占所有甲状腺癌的85％左右)中的发生率为10％～20％，RET融合在年轻患者中更为常见，特别是年轻的非吸烟肺腺癌患者，发生率高达7％～17％。目前对于RET基因改变的治疗方案主要是使用多激酶抑制剂类药物，比如卡博替尼、凡德他尼，由于靶向性不高，通常会发生脱靶导致的VEGFR抑制相关的严重毒性。
蓝图药品公司(Blueprint)和莱德克斯制药公共有限公司(Loxo Oncology)公布了其开发的高效且具有选择性的口服RET抑制剂BLU-667和LOXO-292。Blueprint临床数据结果显示BLU-667表现出广泛的抗肿瘤活性，在具有RET融合和突变的肿瘤患者中总体缓解率(ORR)为45％，其中非小细胞肺癌和甲状腺髓样癌患者的ORR分别为50％和40％。美国FDA授予Loxo Oncology公司的在研药物LOXO-292突破性疗法认定，用于治疗携带RET基因变异的非小细胞肺癌(NSCLC)和甲状腺髓样癌(MTC)患者。
因此，开发新的具有RET激酶抑制活性的、具有更好药效学、药代动力学性能的化合物已成为开发新型抗肿瘤药物的重要研究项目，并最终用于人类肿瘤等疾病的治疗中。
申请公开号为CN113698390A的中国专利公开了用作RET激酶抑制剂的系列化合物，此专利公开的系列化合物具有非常好的RET抑制活性。在该专利申请中公开了一种结构式如式(A)所示的化合物。目前还没有见到有关于该化合物的晶型专利的报道。对式(A)所示化合物进行晶型的筛选开发，找到稳定可靠的晶型物，以保证式(A)所示的化合物的质量稳定性，以便更好的用于药学及药物制剂和将来临床使用，以达到药物质量稳定可控，以及更好的溶出和更高的生物利用度，从而避免由于药物不稳定产生有毒杂质等所导致的副作用等安全性问题，将对后期推进具有重要意义。
发明内容
本发明的目的在于提供一种嘧啶并氮杂环类化合物的晶型、及其制备方法及应用。
本发明的第一方面，提供了一种式A化合物的晶型，所述晶型选自下组：游离碱晶型III、富马酸盐晶型I、苹果酸盐晶型I，
其中，
所述游离碱晶型III的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.8±0.2°、7.3±0.2°、8.2±0.2°、9.3±0.2°、10.4±0.2°、10.7±0.2°、13.0±0.2°、13.3±0.2°、14.7±0.2°、16.7±0.2°、18.4±0.2°、19.0±0.2°、23.4±0.2°；
所述富马酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、8.7±0.2°、10.6±0.2°、14.6±0.2°、15.5±0.2°、17.5±0.2°、18.0±0.2°、19.2±0.2°、27.1±0.2°；
所述苹果酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、9.3±0.2°、10.7±0.2°、11.2±0.2°、11.8±0.2°、15.3±0.2°、17.3±0.2°、17.6±0.2°、18.3±0.2°、19.0±0.2°、23.5±0.2°、25.7±0.2°、28.0±0.2°。
在另一优选例中，
所述游离碱晶型III的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.8±0.2°、7.3±0.2°、7.8±0.2°、8.2±0.2°、9.3±0.2°、10.4±0.2°、10.7±0.2°、11.0±0.2°、13.0±0.2°、13.3±0.2°、14.0±0.2°、14.7±0.2°、15.2±0.2°、16.7±0.2°、17.5±0.2°、18.4±0.2°、19.0±0.2°、21.1±0.2°、22.2±0.2°、23.4±0.2°、29.5±0.2°；
所述富马酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、8.7±0.2°、10.6±0.2°、11.0±0.2°、11.9±0.2°、12.1±0.2°、12.5±0.2°、14.6±0.2°、15.5±0.2°、15.9±0.2°、17.5±0.2°、18.0±0.2°、19.2±0.2°、26.7±0.2°、27.1±0.2°；
所述苹果酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、9.3±0.2°、10.2±0.2°、10.7±0.2°、11.2±0.2°、11.8±0.2°、13.2±0.2°、15.3±0.2°、16.2±0.2°、17.3±0.2°、17.6±0.2°、18.3±0.2°、19.0±0.2°、23.5±0.2°、24.8±0.2°、25.7±0.2°、27.6±0.2°、28.0±0.2°。
在另一优选例中，
所述游离碱晶型III的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.8±0.2°、7.3±0.2°、7.8±0.2°、8.2±0.2°、9.3±0.2°、10.4±0.2°、10.7±0.2°、11.0±0.2°、13.0±0.2°、13.3±0.2°、14.0±0.2°、14.7±0.2°、15.2±0.2°、16.7±0.2°、17.5±0.2°、18.4±0.2°、19.0±0.2°、21.1±0.2°、21.6±0.2°、22.2±0.2°、23.4±0.2°、24.9±0.2°、25.9±0.2°、26.3±0.2°、27.9±0.2°、28.4±0.2°、29.5±0.2°、30.0±0.2°、35.6±0.2°；
所述富马酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、8.2±0.2°、8.7±0.2°、9.9±0.2°、10.6±0.2°、11.0±0.2°、11.9±0.2°、12.1±0.2°、12.5±0.2°、14.6±0.2°、15.5±0.2°、15.9±0.2°、16.5±0.2°、17.5±0.2°、18.0±0.2°、19.2±0.2°、21.2±0.2°、22.2±0.2°、23.9±0.2°、25.5±0.2°、26.7±0.2°、27.1±0.2°、28.8±0.2°；
所述苹果酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、9.3±0.2°、10.2±0.2°、10.7±0.2°、11.2±0.2°、11.8±0.2°、12.1±0.2°、13.2±0.2°、15.3±0.2°、16.2±0.2°、16.8±0.2°、17.1±0.2°、17.3±0.2°、17.6±0.2°、18.3±0.2°、19.0±0.2°、23.5±0.2°、24.8±0.2°、25.2±0.2°、25.7±0.2°、27.6±0.2°、28.0±0.2°、29.7±0.2°。
在另一优选例中，
所述游离碱晶型III的X射线粉末衍射图谱基本如图5所示；
所述富马酸盐晶型I的X射线粉末衍射图谱基本如图37所示；
所述苹果酸盐晶型I的X射线粉末衍射图谱基本如图41所示。
在另一优选例中，所述晶型选自下组：富马酸盐晶型I、苹果酸盐晶型I。
在另一优选例中，所述晶型为富马酸盐晶型I。
在另一优选例中，所述富马酸盐晶型I为三斜晶系，P1空间群。
在另一优选例中，所述富马酸盐晶型I中，每个不对称单元含两个游离碱分子和一个富马酸分子，每个晶胞含一个不对称单元。
在另一优选例中，所述富马酸盐晶型I的晶胞参数是a＝9.7377(2)，α＝105.363°，β＝91.357°，γ＝109.397°， Z＝1。
本发明的第二方面，提供了一种游离碱晶型III的制备方法，所述方法选自下组：
方法一：将式A所示化合物与溶剂m混合，溶清，结晶析出；所述溶剂m选自正丁酮、丙酮和乙腈中的一种；
方法二：将式A所示化合物与溶剂n混合，高温打浆，结晶析出；所述溶剂n选自乙酸乙酯、乙酸异丙酯和甲基叔丁基醚中的一种。
在另一优选例中，所述式A所示化合物为游离碱晶型I。
在另一优选例中，所述“高温”为50-70℃。
本发明的第三方面，提供了一种富马酸盐晶型I或苹果酸盐晶型I的制备方法，所述方法选自下组：
方法一：将式A化合物溶解在有机溶剂中，室温搅拌，然后加入富马酸或苹果酸，继续搅拌，固液分离得到所述富马酸盐晶型I或苹果酸盐晶型I；
方法二：向式A化合物的富马酸盐或苹果酸盐中加入有机溶剂，悬浮搅拌，固液分离得到所述富马酸盐晶型I或苹果酸盐晶型I。
在另一优选例中，所述有机溶剂为2-甲基四氢呋喃。
本发明的第四方面，提供了一种本发明第一方面所述晶型的用途，用于制备药物，所述药物用于选自下组的用途：
1)治疗与RET调节有关的疾病或病症；
2)治疗肿瘤；
3)抑制细胞增殖。
在另一优选例中，所述与RET调节有关的疾病或病症选自下组：甲状腺癌、非小细胞肺癌。
在另一优选例中，所述细胞选自下组：人甲状腺癌细胞和人肺癌细胞。
在另一优选例中，所述肿瘤选自下组：甲状腺癌、非小细胞肺癌。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1是本发明游离碱晶型I的X射线粉末衍射图；
图2是本发明游离碱晶型I的DSC-TGA图谱；
图3是本发明游离碱晶型Ⅱ的X射线粉末衍射图；
图4是本发明游离碱晶型Ⅱ的DSC-TGA图谱；
图5是本发明游离碱晶型Ⅲ的X射线粉末衍射图；
图6是本发明游离碱晶型Ⅲ的DSC-TGA图谱；
图7是本发明游离碱晶型Ⅳ的X射线粉末衍射图；
图8是本发明游离碱晶型Ⅳ的DSC-TGA图谱；
图9是本发明游离碱晶型Ⅴ的X射线粉末衍射图；
图10是本发明游离碱晶型Ⅴ的DSC-TGA图谱；
图11是本发明游离碱晶型Ⅵ的X射线粉末衍射图；
图12是本发明游离碱晶型Ⅵ的DSC-TGA图谱；
图13是本发明游离碱晶型Ⅶ的X射线粉末衍射图；
图14是本发明游离碱晶型Ⅶ的DSC-TGA图谱；
图15是本发明游离碱晶型Ⅷ的X射线粉末衍射图；
图16是本发明游离碱晶型Ⅷ的DSC-TGA图谱；
图17是本发明游离碱晶型Ⅸ的X射线粉末衍射图；
图18是本发明游离碱晶型Ⅸ的DSC-TGA图谱；
图19是本发明盐酸盐晶型I的X射线粉末衍射图；
图20是本发明盐酸盐晶型I的DSC-TGA图谱；
图21是本发明对甲苯磺酸盐晶型I的X射线粉末衍射图；
图22是本发明对甲苯磺酸盐晶型I的DSC-TGA图谱；
图23是本发明甲磺酸盐晶型I的X射线粉末衍射图；
图24是本发明甲磺酸盐晶型I的DSC-TGA图谱；
图25是本发明苯磺酸盐晶型I的X射线粉末衍射图；
图26是本发明苯磺酸盐晶型I的DSC-TGA图谱；
图27是本发明草酸盐晶型I的X射线粉末衍射图；
图28是本发明草酸盐晶型I的DSC-TGA图谱；
图29是本发明草酸盐晶型Ⅱ的X射线粉末衍射图；
图30是本发明草酸盐晶型Ⅱ的DSC-TGA图谱；
图31是本发明马来酸盐晶型I的X射线粉末衍射图；
图32是本发明马来酸盐晶型I的DSC-TGA图谱；
图33是本发明磷酸盐晶型I的X射线粉末衍射图；
图34是本发明磷酸盐晶型I的DSC-TGA图谱；
图35是本发明酒石酸盐晶型I的X射线粉末衍射图；
图36是本发明酒石酸盐晶型I的DSC-TGA图谱；
图37是本发明富马酸盐晶型I的X射线粉末衍射图；
图38是本发明富马酸盐晶型I的DSC-TGA图谱；
图39是本发明柠檬酸盐晶型I的X射线粉末衍射图；
图40是本发明柠檬酸盐晶型I的DSC-TGA图谱；
图41是本发明苹果酸盐晶型I的X射线粉末衍射图；
图42是本发明苹果酸盐晶型I的DSC-TGA图谱。
图43是本发明富马酸盐晶型I的单晶结构图(含两个游离碱分子和一个富马酸分子，每个晶胞含一个不对称单元，碳(C-8/C-36)的绝对构型为S构型)。
图44是本发明游离碱无定型的X射线粉末衍射图。
图45和图46是本发明游离碱晶型I的PLM图。
具体实施方式
本发明人经过长期而深入的研究，意外地制备了一系列具有优异稳定性的式A所示化合物的晶型，尤其是游离碱晶型III、富马酸盐晶型I、苹果酸盐晶型I。在此基础上，发明人完成了本发明。
晶型
晶型是药物存在的固体物质状态，药物晶型研究就是对药物基础状态的研究，只有对化学药物晶型状态有了比较充分和全面的认识，才有可能寻找更合适于治疗疾病的药物晶型固体物质。药物晶型可以影响药物的理化性质，直接影响药物临床发挥治疗疾病作用的基础。同一药物的不同晶型在外观、溶解度、熔点、溶出度、生物有效性等方面可能会有显著不同，从而影响药物的稳定性、生物利用度及疗效。因此研究该化合物的稳定晶型具有重要意义。
同一药物的活性成分一般存在两种或以上的晶体形态，称为药物多晶型。不同晶型具有各自不同的溶解度与溶解速率，通过引起体内生物利用度的变化，来影响药物在临床上的治疗作用。药物晶型的不同，可能会影响其在体内的溶出和吸收，进而影响药物的生物利用度、临床疗效和安全性。同时，药物晶型的稳定性也十分重要。为了提高药物的生物利用度，减少毒性，增进治疗效果就要更加重视药物晶型稳定性。稳定性好的晶型可以保证在制备和贮存过程中药物剂型的物理化学稳定性，保持药物剂型好的溶解度和生物利用度，保证每批药物间的等效性。同一种药物往往具有多种晶型，目前将具有更好的治疗效果，临床上最为适用的晶型称作优势药物晶型。
本发明对式(A)所示的化合物的晶型进行了筛选，尽可能多的找出了原料药的不同晶型。筛选出的晶型通过粉末X射线衍射分析(XRPD)、差示扫描量热法(DSC)、热重分析(TGA)、氢谱液态核磁(1H NMR)和高效液相色谱(HPLC)等手段进行了鉴别，并进一步通过热力学稳定性、引湿性研究考察，确定出了优势药物游离碱晶型III、富马酸盐晶型Ⅰ和苹果酸盐晶型Ⅰ，为后续药物代谢动力学实验与动物学实验的晶型选择提供了参考依据。
经进一步的药代动力学研究试验，本发明提供的式(A)所示的游离碱晶型III、富马酸盐晶型Ⅰ和苹果酸盐晶型Ⅰ在生物体内易于吸收和稳定性好，其在生物体内有很好的血药浓度分布，生物利用度高。同时由于其物理稳定性好，非常有利于在后面的制剂开发中的药物的质量控制及有更长的保质期，从而使药物能够达到更好的疗效。
在另一优选例中，所述式A化合物还具有选自下组的晶型：游离碱晶型I、游离碱晶型II、游离碱晶型IV、游离碱晶型V、游离碱晶型VI、游离碱晶型VII、游离碱晶型VIII、游离碱晶型IX，
其中，所述游离碱晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.8±0.2°、8.6±0.2°、9.9±0.2°、12.9±0.2°、13.7±0.2°、18.8±0.2°；
所述游离碱晶型II的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.4±0.2°、8.1±0.2°、9.7±0.2°、10.0±0.2°、10.5±0.2°、13.6±0.2°；
所述游离碱晶型IV的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.7±0.2°、8.3±0.2°、9.5±0.2°、11.8±0.2°、12.2±0.2°、12.5±0.2°、12.9±0.2°、13.4±0.2°、14.4±0.2°、15.4±0.2°、17.3±0.2°、18.4±0.2°、18.6±0.2°、20.2±0.2°；
所述游离碱晶型V的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.4±0.2°、9.5±0.2°、9.8±0.2°、11.9±0.2°、13.7±0.2°；
所述游离碱晶型VI的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.7±0.2°、7.4±0.2°、8.4±0.2°、9.7±0.2°、10.3±0.2°、11.5±0.2°、12.4±0.2°、14.6±0.2°、15.7±0.2°、17.1±0.2°、17.4±0.2°、18.7±0.2°、20.3±0.2°、23.1±0.2°；
所述游离碱晶型VII的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.6±0.2°、7.0±0.2°、7.4±0.2°、9.5±0.2°、9.9±0.2°、10.3±0.2°、13.0±0.2°、13.3±0.2°、14.4±0.2°、14.9±0.2°、16.1±0.2°、18.1±0.2°、18.6±0.2°、18.9±0.2°、20.6±0.2°；
所述游离碱晶型VIII的X射线粉末衍射图谱在如下2θ值处具有特征峰：5±0.2°、8.3±0.2°、8.6±0.2°、10.1±0.2°、11.2±0.2°、12.4±0.2°、12.6±0.2°、13.1±0.2°、13.6±0.2°、17.7±0.2°、18±0.2°、19.2±0.2°；
所述游离碱晶型IX的X射线粉末衍射图谱在如下2θ值处具有特征峰：7.9±0.2°、11.8±0.2°、15.9±0.2°、16.4±0.2°、17.3±0.2°、20.0±0.2°、20.2±0.2°、21.2±0.2°。
在另一优选例中，所述游离碱晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.8±0.2°、6.8±0.2°、8.6±0.2°、9.9±0.2°、12.0±0.2°、12.9±0.2°、13.7±0.2°、14.4±0.2°、15.2±0.2°、16.5±0.2°、17.2±0.2°、17.6±0.2°、18.4±0.2°、18.8±0.2°、21.9±0.2°、23.0±0.2°、23.5±0.2°；
所述游离碱晶型II的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.4±0.2°、8.1±0.2°、9.7±0.2°、10.0±0.2°、10.5±0.2°、11.8±0.2°、12.6±0.2°、13.6±0.2°、16.4±0.2°、19.8±0.2°、23.0±0.2°；
所述游离碱晶型IV的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.7±0.2°、6.6±0.2°、6.7±0.2°、7.2±0.2°、8.3±0.2°、9.5±0.2°、10.0±0.2°、11.5±0.2°、11.8±0.2°、12.2±0.2°、12.5±0.2°、12.9±0.2°、13.1±0.2°、13.4±0.2°、14.4±0.2°、14.6±0.2°、15.4±0.2°、16.1±0.2°、16.6±0.2°、17.3±0.2°、17.5±0.2°、18.0±0.2°、18.4±0.2°、18.6±0.2°、20.2±0.2°、22.7±0.2°、23.0±0.2°；
所述游离碱晶型V的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.4±0.2°、8.2±0.2°、9.5±0.2°、9.8±0.2°、11.1±0.2°、11.9±0.2°、12.8±0.2°、13.7±0.2°、16.4±0.2°、19.2±0.2°、20.9±0.2°、23.2±0.2°；
所述游离碱晶型VI的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.7±0.2°、6.7±0.2°、7.4±0.2°、8.4±0.2°、9.7±0.2°、10.3±0.2°、11.5±0.2°、12.0±0.2°、12.4±0.2°、13.0±0.2°、13.6±0.2°、14.6±0.2°、15.7±0.2°、16.2±0.2°、17.1±0.2°、17.4±0.2°、18.7±0.2°、20.3±0.2°、21.4±0.2°、22.8±0.2°、23.1±0.2°、29.9±0.2°；
所述游离碱晶型VII的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.6±0.2°、7.0±0.2°、7.4±0.2°、7.8±0.2°、8.7±0.2°、9.5±0.2°、9.9±0.2°、10.3±0.2°、11.7±0.2°、12.1±0.2°、13.0±0.2°、13.3±0.2°、13.8±0.2°、14.4±0.2°、14.9±0.2°、16.1±0.2°、16.3±0.2°、18.1±0.2°、18.6±0.2°、18.9±0.2°、20.6±0.2°、23.5±0.2°；
所述游离碱晶型VIII的X射线粉末衍射图谱在如下2θ值处具有特征峰：4.3±0.2°、5±0.2°、5.5±0.2°、8.3±0.2°、8.6±0.2°、10.1±0.2°、10.4±0.2°、11.2±0.2°、12.4±0.2°、12.6±0.2°、13.1±0.2°、13.6±0.2°、14.4±0.2°、16.6±0.2°、17.7±0.2°、18±0.2°、19.2±0.2°、20.3±0.2°、22±0.2°、23.6±0.2°、24.9±0.2°、25.1±0.2°、26.1±0.2°、26.4±0.2°；
所述游离碱晶型IX的X射线粉末衍射图谱在如下2θ值处具有特征峰：7.9±0.2°、8.4±0.2°、11.8±0.2°、13.8±0.2°、15.9±0.2°、16.4±0.2°、17.3±0.2°、19.0±0.2°、19.2±0.2°、20.0±0.2°、20.2±0.2°、21.2±0.2°、28.4±0.2°。
在另一优选例中，所述游离碱晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.8±0.2°、6.8±0.2°、8.6±0.2°、9.9±0.2°、12.0±0.2°、12.9±0.2°、13.7±0.2°、14.4±0.2°、15.2±0.2°、16.5±0.2°、17.2±0.2°、17.6±0.2°、18.1±0.2°、18.4±0.2°、18.8±0.2°、19.5±0.2°、20.5±0.2°、21.6±0.2°、21.9±0.2°、22.3±0.2°、23.0±0.2°、23.5±0.2°、25.1±0.2°、25.7±0.2°、26.1±0.2°、29.6±0.2°、29.9±0.2°；
所述游离碱晶型II的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.4±0.2°、6.6±0.2°、8.1±0.2°、9.1±0.2°、9.7±0.2°、10.0±0.2°、10.5±0.2°、11.4±0.2°、11.8±0.2°、12.6±0.2°、13.6±0.2°、14.6±0.2°、16.4±0.2°、16.8±0.2°、19.2±0.2°、19.8±0.2°、21.1±0.2°、21.7±0.2°、22.0±0.2°、23.0±0.2°、23.8±0.2°、25.6±0.2°、29.4±0.2°；
所述游离碱晶型IV的X射线粉末衍射图谱在如下2θ值处具有特征峰：3.5±0.2°、4.1±0.2°、5.3±0.2°、5.7±0.2°、6.6±0.2°、6.7±0.2°、7.2±0.2°、8.3±0.2°、9.5±0.2°、10.0±0.2°、10.2±0.2°、10.7±0.2°、11.5±0.2°、11.8±0.2°、12.2±0.2°、12.5±0.2°、12.9±0.2°、13.1±0.2°、13.4±0.2°、14.4±0.2°、14.6±0.2°、15.4±0.2°、15.6±0.2°、16.1±0.2°、16.6±0.2°、16.7±0.2°、17.3±0.2°、17.5±0.2°、18.0±0.2°、18.4±0.2°、18.6±0.2°、18.9±0.2°、19.7±0.2°、19.9±0.2°、20.2±0.2°、20.7±0.2°、21.0±0.2°、21.1±0.2°、21.9±0.2°、22.2±0.2°、22.7±0.2°、23.0±0.2°、23.6±0.2°、24.5±0.2°、25.2±0.2°、25.4±0.2°、25.9±0.2°、26.1±0.2°、27.5±0.2°；
所述游离碱晶型V的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.4±0.2°、8.2±0.2°、9.5±0.2°、9.8±0.2°、11.1±0.2°、11.9±0.2°、12.8±0.2°、13.7±0.2°、14.9±0.2°、16.4±0.2°、17.1±0.2°、18.7±0.2°、19.2±0.2°、20.3±0.2°、20.9±0.2°、22.0±0.2°、22.4±0.2°、23.2±0.2°、25.3±0.2°、25.8±0.2°、26.4±0.2°、26.9±0.2°、28.1±0.2°、29.0±0.2°、30.1±0.2°；
所述游离碱晶型VI的X射线粉末衍射图谱在如下2θ值处具有特征峰：4.2±0.2°、5.7±0.2°、6.7±0.2°、7.4±0.2°、8.4±0.2°、9.7±0.2°、10.3±0.2°、11.5±0.2°、12.0±0.2°、12.4±0.2°、13.0±0.2°、13.6±0.2°、14.6±0.2°、15.7±0.2°、16.2±0.2°、17.1±0.2°、17.4±0.2°、18.7±0.2°、20.3±0.2°、21.0±0.2°、21.4±0.2°、22.8±0.2°、23.1±0.2°、25.8±0.2°、26.3±0.2°、27.2±0.2°、28.8±0.2°、29.2±0.2°、29.9±0.2°；
所述游离碱晶型VII的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.6±0.2°、7.0±0.2°、7.4±0.2°、7.8±0.2°、8.7±0.2°、9.5±0.2°、9.9±0.2°、10.3±0.2°、11.4±0.2°、11.7±0.2°、12.1±0.2°、13.0±0.2°、13.3±0.2°、13.8±0.2°、14.4±0.2°、14.9±0.2°、15.5±0.2°、16.1±0.2°、16.3±0.2°、16.9±0.2°、18.1±0.2°、18.6±0.2°、18.9±0.2°、19.7±0.2°、20.6±0.2°、21.2±0.2°、21.8±0.2°、22.5±0.2°、23.1±0.2°、23.5±0.2°、24.9±0.2°、25.4±0.2°；
所述游离碱晶型VIII的X射线粉末衍射图谱在如下2θ值处具有特征峰：4.3±0.2°、5±0.2°、5.5±0.2°、6.7±0.2°、8.3±0.2°、8.6±0.2°、9.7±0.2°、10.1±0.2°、10.4±0.2°、11.2±0.2°、12.4±0.2°、12.6±0.2°、13.1±0.2°、13.6±0.2°、14.4±0.2°、15±0.2°、15.6±0.2°、16.6±0.2°、17.7±0.2°、18±0.2°、18.6±0.2°、19.2±0.2°、20.3±0.2°、21±0.2°、21.3±0.2°、22±0.2°、22.6±0.2°、23.1±0.2°、23.6±0.2°、24.9±0.2°、25.1±0.2°、25.7±0.2°、26.1±0.2°、26.4±0.2°、27±0.2°、27.5±0.2°、28.9±0.2°、30.3±0.2°、31.8±0.2°；
所述游离碱晶型IX的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.5±0.2°、5.8±0.2°、6.8±0.2°、7.9±0.2°、8.4±0.2°、8.6±0.2°、9.8±0.2°、11.8±0.2°、13.3±0.2°、13.8±0.2°、14.0±0.2°、15.9±0.2°、16.4±0.2°、17.3±0.2°、17.6±0.2°、19.0±0.2°、19.2±0.2°、20.0±0.2°、20.2±0.2°、20.6±0.2°、21.2±0.2°、24.2±0.2°、27.8±0.2°、28.4±0.2°、29.4±0.2°。
在另一优选例中，所述游离碱晶型I的X射线粉末衍射图谱基本如图1所示；
所述游离碱晶型II的X射线粉末衍射图谱基本如图3所示；
所述游离碱晶型IV的X射线粉末衍射图谱基本如图7所示；
所述游离碱晶型V的X射线粉末衍射图谱基本如图9所示；
所述游离碱晶型VI的X射线粉末衍射图谱基本如图11所示；
所述游离碱晶型VII的X射线粉末衍射图谱基本如图13所示；
所述游离碱晶型VIII的X射线粉末衍射图谱基本如图15所示；
所述游离碱晶型IX的X射线粉末衍射图谱基本如图17所示。
与现有技术相比，本发明具有以下主要优点：
1)所述晶型具有优异的稳定性、高的溶解度；
2)相比于游离碱晶型I，本发明游离碱晶型Ⅲ、富马酸盐晶型I、苹果酸盐晶型I具有更优的药代性质，更高的动物体内暴露量。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。
除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。
本发明中缩写见下表
化学试剂缩写：
2-Me-THF 2-甲基-四氢呋喃
ACN      乙腈
DCM      二氯甲烷
DMSO     二甲亚砜
EA       乙酸乙酯
EtOH     乙醇
Hept     正庚烷
IPA      异丙醇
IPAc     乙酸异丙酯
MEK      正丁酮
MeOH     甲醇
MTBE     甲基叔丁基醚
n-BuOH   正丁醇
THF      四氢呋喃
Tol      甲苯
HCL      盐酸
H2SO4     硫酸
TS       对甲苯磺酸
MS       甲磺酸
BS       苯磺酸
OX       草酸
MA       马来酸
H3PO4     磷酸
TA       L-酒石酸
FU       富马酸
CI       柠檬酸一水合物
其它缩写(按字母顺序排列)：
DSC        差示扫描量热
DVS        动态水蒸气吸附
HPLC       高效液相色谱
NMR        核磁共振质谱
PLM        偏光显微镜
RT         室温(20–25℃)
TGA           热重分析
XRPD          X-射线粉末衍射
空腹模拟肠液  FaSSIF
饱腹模拟肠液  FeSSIF
模拟胃液      Simulated Gastric Fluid
下面对本发明的技术方案进行详细说明。
一、式(A)所示化合物的合成
制备方法同专利CN113698390A，合成路线如下：
具体步骤如下所述：
化合物7的合成
将对甲氧基苯甲醇(16.0g,116.4mmol)，叔丁醇钾(24.9g,222.2mmol)溶解于400mL无水二氧六环中，冰浴下，将溶于300mL二氧六环的化合物6(20.0g，105.8mmol)滴加到反应瓶中，磁力搅拌2h溶液逐渐变成胶体状，反应完毕后加水淬灭，乙酸乙酯萃取水相，分离出有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压旋干得粗品，再用石油醚打浆两小时，抽滤，真空干燥得到28.8g化合物7，MS[M+H]291。
1H NMR(400MHz,Chloroform-d)δ8.07(s,1H),7.47(d,J＝8.7Hz,2H),6.95(d,J＝8.7Hz,2H),5.56(s,2H),3.84(s,3H).
化合物8的合成
将化合物7(2.0g，6.80mmol)溶于60mL无水四氢呋喃中，依次加入乙醇(1.1mL，17.2mmol)和三苯基膦(2.9g，11.0mmol)，将反应液氮气保护，冰浴下加入偶氮二甲酸二异丙酯(2.8g,13.6mmol),自然升温至室温并搅拌3h，加入饱和食盐水淬灭反应，然后用乙酸乙酯萃取，分离出有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得粗品，经柱层析纯化得到1.78g化合物8，MS[M+H]319。
1H NMR(400MHz,Chloroform-d)δ7.99(s,1H),7.45(d,J＝8.8Hz,2H),6.94(d,J＝8.7Hz,2H),5.54(s,2H),4.45(q,J＝7.2Hz,2H),3.83(s,3H),1.50(t,J＝7.3Hz,3H).
化合物10的合成
将化合物8(1.78g，5.60mmol)和化合物9(1.82g，6.16mmol)溶于60mL 1,4-二氧六环和12mL水混合溶剂中，依次加入醋酸钯(126mg，0.56mmol)，1,1'-双(二苯基膦)二茂铁(621mg，1.12mmol)和碳酸钾(1.16g，8.40mmol)，将反应液氮气保护下90℃搅拌过夜。反应结束后，冷却至室温，减压浓缩，乙酸乙酯萃取，合并有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，柱层析纯化得到1.52g化合物10，MS[M+H]453。
1H NMR(400MHz,Chloroform-d)δ7.93(s,1H),7.42(s,2H),7.35-7.29(m,1H),6.91(d,J＝8.7Hz,2H),5.55(s,2H),4.46(q,J＝7.3Hz,2H),3.81(d,J＝2.9Hz,6H),3.33(s,3H),2.97–2.49(m,4H),2.41–2.20(m,1H),2.20–2.08(m,1H),1.50(t,J＝7.2Hz,3H).
化合物11的合成
将化合物10(1.52g，3.36mmol)溶于15mL乙酸乙酯和15mL甲醇组成的混合溶剂中，然后加入湿钯碳(152mg)，氢气氛围下室温搅拌过夜。反应结束后，将反应液通过硅藻土过滤，甲醇洗涤，滤液减压浓缩得到1.33g化合物11，MS[M+H]335。
1H NMR(400MHz,Chloroform-d)δ10.64(s,1H),8.00(s,1H),4.33(q,J＝7.1Hz,2H),3.73(s,3H),3.24(s,3H),2.71–2.60(m,1H),2.14(d,J＝12.4Hz,2H),1.97–1.72(m,6H),1.44(t,J＝7.2Hz,3H).
化合物12的合成
将化合物11(1.33g，粗品)溶于30mL三氯氧磷中，氮气保护下，100℃搅拌3h。反应结束后减压蒸去溶剂，而后加入碎冰，饱和碳酸氢钠溶液调至中性，乙酸乙酯萃取，合并有机相用饱和食盐水洗涤，无水硫酸钠干燥有机相，减压浓缩，柱层析纯化得到936mg化合物12，MS[M+H]353。
1H NMR(400MHz,Chloroform-d)δ8.00(s,1H),4.46(q,J＝7.3Hz,2H),3.73(s,3H),3.24(s,3H),3.09–2.84(m,1H),2.13(d,J＝11.8Hz,2H),2.00–1.72(m,6H),1.47(t,J＝7.3Hz,3H).
化合物14的合成
将化合物12(936mg，2.66mmol)和化合物13(284mg，2.92mmol)溶于15mL二甲基亚砜中，然后加入二异丙基乙胺(686mg，5.32mmol)，氮气保护下100℃搅拌过夜。反应结束后，冷却至室温，乙酸乙酯稀释，有机相用饱和食盐水洗涤，无水硫酸钠干燥有机相，减压浓缩，柱层析纯化得到728mg化合物14，MS[M+H]414。
1H NMR(400MHz,Chloroform-d)δ7.78(s,1H),6.54(s,1H),4.44(q,J＝6.3,5.3Hz,2H),3.80(s,3H),3.33(s,3H),2.93–2.82(m,1H),2.36(s,J＝6.5Hz,3H),2.23–1.62(m,8H),1.49(t,J＝7.2Hz,3H).
化合物int-A的合成
将化合物14(728mg，1.76mmol)溶于16mL甲醇和2mL水组成的混合溶剂中加入氢氧化锂一水合物(222mg，5.29mmol)，60℃搅拌过夜。反应结束后，将反应液冷却至室温，减压蒸去甲醇，DCM打浆，抽滤且无需纯化，直接用于下一步，共得到797mg粗品int-A，MS[M+H]400。
化合物2的合成
50mL单口瓶中加入化合物1(1.57g，10mmol)、4-氟-1H吡唑(860mg，10mmol)、N,N-二甲基甲酰胺(10mL)、碳酸钾(2.76g，20mmol)，将反应置于100℃下反应8h。LC-MS显示反应完毕，搅拌下将反应液倒入水(200mL)中有固体析出，过滤，水洗滤饼，将滤饼干燥，得1.08g化合物2。[M+1]:207.03。
化合物3的合成
100mL单口瓶中依次加入化合物2(870mg,4.2mmol)、R-叔丁基亚磺酰胺(560mg,4.63mmol)、钛酸四乙酯(1.92g,8.4mmol)、干燥四氢呋喃(10mL)，将反应置于75℃下反应过夜。冷却，加水(30mL)稀释，抽滤，滤液用乙酸乙酯(20mL)萃取三次，无水硫酸钠干燥有机相，减压蒸干，柱层析得产品500mg。
化合物4的合成
50mL单口瓶中依次加入化合物3(100mg,0.32mmol)、干燥四氢呋喃(2mL)，降温至-78℃，滴加三仲丁基硼氢化锂(0.7mL,0.7mmol，1.0M)，于-78℃下反应1h，-60℃下加甲醇(1mL)淬灭反应，加水(5mL)稀释，用乙酸乙酯(5mL)萃取三次，干燥有机相，减压蒸干，柱层析得产物93mg。
1H NMR(400MHz,CDCl3)δ9.23(d,J＝1.4Hz,1H),8.36(d,J＝1.5Hz,1H),8.33(d,J＝4.6Hz,1H),7.66(d,J＝4.3Hz,1H),4.77(p,J＝6.6Hz,1H),3.74(d,J＝5.9Hz,1H),1.67(d,J＝6.8Hz,3H),1.22(s,9H)。
化合物5的合成
50mL单口瓶中依次加入化合物4(100mg，0.32mmol)、二氯甲烷(3mL)，甲醇(0.5mL)、氯化氢/二氧六环(4M,1.5mL)将反应置于室温下反应3h。LC-MS检测反应完全，加二氯甲烷稀释，过滤，乙酸乙酯洗涤滤饼，干燥滤饼得产物67mg。[M+1]:208.06。
式(A)的合成
将化合物int-A(270mg,0.68mmol)和化合物5(165mg，0.68mmol)溶于9mL N,N-二甲基甲酰胺中，加入二异丙基乙胺(699mg，5.41mmol)，室温搅拌15min后，再加入HATU(386mg,1.02mmol)，继续室温搅拌1h。将反应液用乙酸乙酯稀释，有机相用饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经高效液相色谱分离纯化得到式(A),Ms[M+H]589和式(B),MS[M+H]589.
式(A)的核磁：1H NMR(400MHz,Chloroform-d)δ9.23(d,J＝1.4Hz,1H),8.37(d,J＝1.4Hz,1H),8.34(d,J＝4.6Hz,1H),7.78(s,1H),7.66(d,J＝4.3Hz,2H), 5.32(p,J＝6.8Hz,1H),4.44(q,J＝7.2Hz,2H),3.32(s,3H),2.93–2.77(m,1H),2.38(s,3H),2.13–1.90(m,8H),1.56(d,J＝6.9Hz,3H),1.48(t,J＝7.2Hz,3H).
将所得式(A)固体进行XRPD表征，结果如图1所示，所得固体为游离碱晶型I。
所述游离碱晶型I作为原料用于下文的晶型制备和盐型制备实验。
二、式(A)所示化合物晶型的制备及评估
室温下，在17种溶剂中通过目测法对游离碱晶型I的溶解度进行评估。将5mg的游离碱晶型I称重至8mL的瓶子中，逐步往里滴加溶剂直到药物溶清或者达到1600V，粗算溶解度(mg/mL)。结果如表1所示。游离碱晶型I在DMSO和THF中显示较高的溶解度(>40mg/mL)，在剩余的溶剂中溶解度差。
表1游离碱晶型I的粗测溶解度结果
如果未溶解则标为“<”，如果溶清则标记为“>”。
1、晶型筛选方法
对式(A)化合物的固体晶型筛选，是从选定的单一溶剂及其混合溶剂结晶或沉淀，来获得亚稳态和稳定态晶型。采用的方法包括挥发结晶、打浆结晶、冷却结晶、反溶剂沉淀。
1.1挥发结晶
根据溶解度数据，在选定的溶剂中进行挥发结晶。在室温或50℃条件下，将5 mg的游离碱晶型I在RT或者50℃搅拌20min溶于溶剂(0.5mL)形成澄清液。溶液平分两部分，一部分用氮气吹干做快速挥发，另一部分覆盖有扎孔的薄膜室温下缓慢挥发。
1.2打浆结晶
将30mg的游离碱晶型I称重至样品瓶，加溶剂形成悬浊液，在RT或50℃下搅拌7天，或70℃下搅拌2天。过滤收集固体，滤饼进行XRPD表征。
1.3冷却结晶
将15mg的游离碱晶型I称重至样品瓶，缓慢加溶剂(0.5mL)，50℃或40℃(DCM)搅拌10分钟。溶液接近澄清，微孔过滤得到母液。溶液平均分两部分，一部分母液放置于冰箱(～4℃)做快速冷却。另一部分放置于加热的金属模块里自然冷却至室温。若固体析出立马表征。
1.4反溶剂沉淀
室温下，将15～25mg的游离碱晶型I室温下溶于THF(0.5mL)或DMSO(0.25mL)中，过滤得到澄清液。逐步往里加反溶剂，每次0.1–1mL，直到固体析出或加入的溶剂体积为10V。若沉淀析出，立马进行表征。
2、晶型分析方法
2.1偏光显微镜分析(PLM)
采用ECLIPSE LV100POL(Nikon,JPN)偏光显微镜进行PLM分析。将少量的样品平铺至载玻片上，滴入香柏油将样品分散后，加上盖玻片。随后，将样品置于显微镜下用4-20倍物镜观察。
2.2 X-射线粉末衍射(XRPD)
使用X射线衍射仪对固体样品进行检测。将样品平铺于零背景单晶硅样品盘上，将样品轻压铺平后，根据表2中参数进行分析。
表2 XRPD测试参数

2.3热重分析(TGA)
样品的TGA数据使用TA仪器进行收集。将1-5mg样品置于已去皮的开口铝制样品盘中，按照表3中仪器参数对样品进行加热测试，并使用TRIOS软件对数据进行分析处理。
表3.TGA测试参数
2.4差示扫描量热分析(DSC)
样品的DSC数据使用TA仪器进行收集。将1-3mg样品置于已扎孔的DSC样品盘中，按照表4中的参数进行加热测定，并使用TRIOS软件对数据进行分析处理。
表4.DSC测试参数
3、晶型分析结果
3.1游离碱晶型I
在MeOH、EtOH和IPA体系室温和50℃高温打浆，及IPA/THF体系中反溶剂沉淀得到的都是游离碱晶型I。
游离碱晶型I的XRPD结果如图1所示，DSC和TGA结果如图2所示，1H-NMR显示该样品含有5.4％的乙醇、0.8％的正庚烷及未知溶剂残留，同时TGA显示80℃之前有1.5％的失重，80-200℃有5.2％的失重，归于溶剂的脱去；DSC显示250℃前有多个吸热峰，归于脱溶剂和样品熔融。
从图1可知，游离碱晶型I的X-射线粉末衍射峰数据如表5所示。
表5
3.2游离碱晶型II
将游离碱晶型Ⅰ溶于三氟乙酸，乙腈和水体系，得到的溶液用碳酸氢钠调至碱性，将有机溶剂减压浓缩后，悬浊液过滤，将固体用水洗涤，得到游离碱晶型II。
1H-NMR显示样品含有0.2％的正庚烷残留，而TGA显示70℃之前含有1.8％的失重，70–145℃含有2.5％的失重，归于水或溶剂的脱去。DSC曲线显示200℃之前有多个吸热峰，归于脱溶剂和样品熔融。因此，推测游离碱晶型II为水合物晶型。使用Cu-Kα射线测量得到的X射线粉末衍射图(XRPD)、DSC-TGA图谱见图3、图4。
从图3可知，游离碱晶型II的X-射线粉末衍射峰数据如表6所示。
表6

3.3游离碱晶型III
在正丁酮、丙酮和乙腈的室温或50/70℃高温下，以及乙酸乙酯、乙酸异丙酯和甲基叔丁基醚的50/70℃高温下打浆得到的都是游离碱晶型III。
游离碱晶型III为高结晶度的团聚微小晶体。游离碱晶型III的XRPD结果如图5所示，DSC和TGA结果如图6所示。TGA显示150℃之前没有明显的失重。1H-NMR显示该样品含有0.03％的丙酮残留。DSC曲线上84℃处有小的吸热峰(加热至100℃晶型未改变)。在245℃有另一个吸热峰，为化合物的熔点。因此，游离碱晶型III是无水晶型。
从图5可知，游离碱晶型III的X-射线粉末衍射峰数据如表7所示。
表7

3.4游离碱晶型IV
在2-Me-THF中快速/慢速挥发结晶，或THF快速挥发都制备得到晶型IV。
游离碱晶型IV是低结晶度的团聚小晶体。游离碱晶型IV的XRPD结果如图7所示，DSC和TGA结果如图8所示。TGA显示在210℃前有36.9％的失重，归于脱溶剂。DSC显示250℃之前有多个吸热峰。加热至170℃后变为无定型。因此，晶型IV是水合物或溶剂化合物。
从图7可知，游离碱晶型IV的X-射线粉末衍射峰数据如表8所示。
表8

3.5游离碱晶型V
在MEK或DCM的体系缓慢挥发，DCM的打浆或冷却结晶中得到游离碱晶型V。
对从MEK体系中得到样品进行表征。游离碱晶型V是具有中等结晶度的针状 晶体。游离碱晶型V的XRPD结果如图9所示，DSC和TGA结果如图10所示。TGA曲线显示60-170℃有5.2％的失重，归于脱溶剂。DSC显示250℃之前有多个吸热峰，加热至175℃后游离碱晶型V转换为无定型。因此，游离碱晶型V是水合物或溶剂化合物。
从图9可知，游离碱晶型V的X-射线粉末衍射峰数据如表9所示。
表9
3.6游离碱晶型VI
游离碱晶型VI从THF的缓慢挥发中制备得到，并对其进行表征。游离碱晶型VI是具有中等结晶度的团聚小晶体。DSC和TGA结果如图12所示。TGA显示90℃之前有2.6％的失重，90-195℃有5.5％的失重，归于溶剂的脱去。游离碱晶型VI加热至195℃后转化为无定型。DSC检测出250℃之前有多个吸热峰，因此，游离碱晶型VI可能是溶剂化合物或水合物。
从图11可知，游离碱晶型VI的X-射线粉末衍射峰数据如表10所示。
表10
3.7游离碱晶型VII
游离碱晶型VII通过在2-Me-THF体系在室温或50℃高温下打浆7天得到。游离碱晶型VII是具有中等结晶度的团聚小颗粒晶体。1H-NMR检测出该样品含有0.97％的2-Me-THF残留。DSC和TGA结果如图14所示。TGA显示在85-240℃之间有1.7％的失重，归于溶剂的脱去。DSC在230℃检测出两个重叠的吸热峰，归为熔点。游离碱晶型VII加热至210℃后晶型未发生改变。因此，游离碱晶型VII是无水晶型。
从图13可知，游离碱晶型VII的X-射线粉末衍射峰数据如表11所示。
表11
3.8游离碱晶型VIII
在50℃水中打浆7天得到游离碱晶型VIII。样品湿样和干样的XRPD谱图不完全一致，不同状态下游离碱晶型VIII的峰发生一定的偏移。DSC和TGA结果如图16所示。样品在40℃干燥后，TGA结果显示160℃之前仍然有超过3.2％的失重。DSC显示250℃之前有多个吸热峰。1H-NMR没有检测出明显的溶剂残留。化合物加热到180℃晶型转化为无定型，因此，游离碱晶型VIII可能是水合物。
从图15可知，干燥的游离碱晶型VIII的X-射线粉末衍射峰数据如表12所示
表12
3.9游离碱晶型IX
从DMSO/H2O的反溶剂沉淀中得到游离碱晶型IX。游离碱晶型IX是中等结晶 度的小颗粒晶体。1H-NMR检测出11.54％的DMSO残留。DSC和TGA结果如图18所示。TGA显示85℃之前有2.1％的失重，85-260℃有11.5％的失重，归于溶剂的脱去。DSC显示在37℃处有一个宽的吸热峰，140℃处有另一个吸热峰。因此，游离碱晶型IX可能是DMSO溶剂化合物。
从图17可知，游离碱晶型IX的X-射线粉末衍射峰数据如表13所示。
表13
3.10游离碱无定型
称取游离碱晶型Ⅰ样品，在室温下将其溶解在20mL二氯甲烷中，过滤后将滤液通过旋蒸快速除掉溶剂，得到的固体样品进行相应的表征测试，其XRD谱图如图44所示。
4游离碱晶型研究结论：
表14
式(A)所示结构的化合物在多晶型筛选中显示出相对复杂的多晶型现象，共得到9种晶型，分别为晶型I-IX。其中两种是无水晶型(晶型III和晶型VII)，且晶型III相对稳定，具有更高的熔点和焓值，而其它都是溶剂化合物或水合物，数据汇总见表14。
在本发明中，X-射线粉末衍射峰的2θ角度值，其误差范围为±0.2°。应该理解，X射线粉末衍射图的2θ值可在机器之间和样品之间稍有变化，其数值范围可能相差在±0.2个单位内，因此所引用的数值不能解释为绝对值。
游离碱晶型III是具有高熔点(245℃)的无水晶型，不同溶剂或方法得到的游离碱晶型III样品都是小颗粒晶体。
本发明提供的游离碱晶型III理化性质稳定，非常适合在接下来制剂等药物研发方面的应用。
本发明提供的晶型Ⅰ-晶型Ⅸ，可用于制备RET抑制剂药物，尤其是晶型III。
三、式(A)所示化合物酸式盐的制备及表征
本发明盐型筛选中所使用的12种酸如表15所示。
表15酸列表
室温下将30mg的游离碱(即上文所制得游离碱晶型I)加入到不同溶剂中，再加入酸溶液或固体酸(1.1eq.或0.6eq.)形成结晶态盐。若无固体析出，尝试不同的结晶方式来制备晶型盐，如冷却结晶、挥发结晶和反溶剂沉淀。过滤收集固体，真空干燥并进一步分析。
1、酸式盐晶型的制备和表征
1.1盐酸盐晶型I
室温下，约30mg的游离碱晶型I加入到丙酮(1.0ml)中。将盐酸溶液(1.1eq.，0.56mol/L，浓盐酸溶于对应的0.1mL溶剂)加入到此悬浊液。加HCl溶液后，悬浊液变黄色胶状。10分钟后，析出固体继续搅拌2天，固液分离得到盐酸盐晶型I。
盐酸盐晶型I是具有中等结晶度的团聚细小晶体。1H-NMR显示样品没有明显的有机溶剂残留。DSC和TGA结果如图20所示。TGA显示150℃之前有1.7％的失重，为水的脱去。DSC加热初始阶段有1个宽的吸热峰，在173℃和218℃检测出另两个吸热峰。因此，盐酸盐晶型I可能是水合物。
其粉末X射线衍射图如图19所示，数据如表16所示：
表16

1.2对甲苯磺酸盐晶型I
室温下，约30mg的游离碱晶型I加入到丙酮(1.0ml)中。加对甲苯磺酸溶液后，悬浊液变成几乎黄色的澄清液。5分钟后，变胶状。1小时后，固体析出继续搅拌2天，固液分离得到对甲苯磺酸盐晶型I。
对甲苯磺酸盐晶型I具有高结晶度的团聚细小晶体。1H-NMR检测出3.0％的丙酮残留，酸碱的摩尔比为1:1。DSC和TGA结果如图22所示。TGA结果显示210℃之前有3.0％的失重，为丙酮的脱去。DSC结果显示在163℃处有一个宽的吸热峰，在212℃处有另一个吸热峰。因此，对甲苯磺酸盐晶型I可能是溶剂化合物。
其粉末X射线衍射图如图21所示，数据如表17所示：
表17
1.3甲磺酸盐晶型I
室温下，约30mg的游离碱晶型I加入到丙酮(1.0ml)中。加甲磺酸溶液后，悬浊液变成黄色的胶状体。1小时后固体析出，继续搅拌2天固液分离得到甲磺酸盐晶型I。
甲磺酸盐晶型I是具有中等结晶度的团聚细小晶体。1H-NMR显示没有明显的有机溶剂残留，酸碱摩尔比为1:1。DSC和TGA结果如图24所示。TGA结果显示在100℃之前有0.4％的失重，归为水的脱去。DSC结果显示在154℃处有一个小的吸热峰，在192℃有另一个吸热峰，由样品熔融导致。因此，甲磺酸盐晶型I可能是无水晶型。
其粉末X射线衍射图如图23所示，数据如表18所示：
表18
1.4苯磺酸盐晶型I
室温下，约30mg的游离碱晶型I加入到丙酮(1.0ml)中。加苯磺酸溶液后，悬浊液变为几乎黄色的澄清液。30秒后，析出大量的固体，补加丙酮(0.5mL)。继续搅拌2天，固液分离得到苯磺酸盐晶型I。
苯磺酸盐晶型I是具有高结晶度的团聚细小晶体。1H-NMR显示有0.19％的丙酮残留，酸碱摩尔比为1:1。DSC和TGA结果如图26所示。TGA结果显示200℃之前没有明显的失重。DSC结果显示235℃处有一个吸热峰，为化合物熔融。因此，苯磺酸盐晶型I是无水晶型。
其粉末X射线衍射图如图25所示，数据如表19所示：
表19
1.5草酸盐晶型I和草酸盐晶型Ⅱ
室温下，约30mg的游离碱晶型I加入到甲醇(1.0ml)中。加入草酸溶液后，悬浊液继续搅拌2天，固液分离得到草酸盐晶型I。
草酸盐晶型I是具有高结晶度的小颗粒晶体。1H-NMR显示没有明显的甲醇残留。DSC和TGA结果如图28所示。TGA结果显示在55-85℃之间有0.2％的失重，归为水的脱去。DSC显示在43℃和81℃处有两个小的吸热峰，在229℃有另一个吸热峰，归为水的脱去和熔融。因此，草酸盐晶型I可能是无水晶型。
草酸盐晶型I的粉末X射线衍射图如图27所示，数据如表20所示：
表20

室温下，约30mg的游离碱晶型I加入到丙酮(1.0ml)中。加入草酸溶液(1.1eq.，0.56mol/L，草酸溶于对应的0.1mL溶剂)后，悬浊液变成黄色胶状体。2分钟后，大量固体析出，补加丙酮(0.5mL)，继续搅拌2天，固液分离得到草酸盐晶型II。
草酸盐晶型II是具有高结晶度的团聚细小晶体。1H-NMR显示有0.07％的丙酮残留。DSC和TGA结果如图30所示。TGA显示在200℃之前没有明显的失重。DSC显示在227℃处有个吸热峰，由样品熔融导致。因此，草酸盐晶型II是无水晶型。
草酸盐晶型II的粉末X射线衍射图如图29所示，数据如表21所示：
表21
1.6马来酸盐晶型I
室温下，约30mg的游离碱晶型I加入到2-甲基四氢呋喃(1.0ml)中。加入马来酸溶液(1.1eq.，0.56mol/L，马来酸溶于对应的0.1mL溶剂)后，悬浊液继续搅拌2天，固液分离得到马来酸盐晶型I。
马来酸盐晶型I具有高结晶度的小颗粒晶体。1H-NMR显示样品含0.41％的2-Me-THF残留，酸碱的摩尔比为1:1。DSC和TGA结果如图32所示。TGA结果显示150℃之前没有明显的失重。DSC显示在175℃有个吸热峰，为化合物熔点。因此，马来酸盐晶型I是无水晶型。
马来酸盐晶型I的粉末X射线衍射图如图31所示，数据如表22所示：
表22
1.7磷酸盐晶型I
室温下，约30mg的游离碱晶型I加入到所选溶剂中。将磷酸溶液(1.1eq.，0.56mol/L，磷酸溶于对应的0.1mL溶剂)加到此悬浊液中，实验过程和结果表23所示。
表23磷酸盐的制备
磷酸盐晶型I为具有高结晶度的微小晶体。1H-NMR显示约0.06％的丙酮残留。DSC和TGA结果如图34所示。TGA结果显示200℃之前没有明显的失重。DSC显示在221℃有个吸热峰，归为化合物熔点。因此，磷酸盐晶型I可能是无水晶型。
磷酸盐晶型I的粉末X射线衍射图如图33所示，数据如表24所示：
表24

1.8酒石酸盐晶型I
室温下，约30mg的游离碱晶型I加入到所选溶剂中。将酒石酸固体(1.1eq.)加到此悬浊液中。实验过程和结果如表25所示。
表25酒石酸盐的制备
酒石酸盐晶型I是具有中等结晶度的团聚细小晶体。1H-NMR显示约0.08％的丙酮残留，酸碱摩尔比为1:1。DSC和TGA结果如图36所示。TGA结果显示在105℃之前有1.7％的失重，在105-165℃之间有0.6％的失重。DSC显示在40℃有1个宽的吸热峰，在146℃和181℃有另两个吸热峰，归为脱水。因此，酒石酸盐晶型I可能是水合物。
酒石酸盐晶型I的粉末X射线衍射图如图35所示，数据如表26所示：
表26
1.9富马酸盐晶型I
室温下，约30mg的游离碱晶型I加入到2-Me-THF中，将富马酸固体(1.1eq.)加到此悬浊液中。实验过程和结果如表27所示。
表27富马酸盐的制备
富马酸盐晶型I具有高结晶度的团聚细小晶体。1H-NMR显示酸碱摩尔比为1:2。DSC和TGA结果如图38所示。TGA结果显示在200℃之前没有明显的失重。DSC显示在239℃有个吸热峰，归为化合物熔点。因此，富马酸盐晶型I为无水晶型。
富马酸盐晶型I的粉末X射线衍射图如图37所示，数据如表28所示：
表28
1.10柠檬酸盐晶型I
室温下，约30mg的游离碱晶型I加入到丙酮(1.0ml)中。将柠檬酸固体(1.1eq.)加到此悬浊液中，继续搅拌2天，固液分离得到柠檬酸盐晶型I。
柠檬酸盐晶型I具有中等结晶度的团聚细小晶体。1H-NMR显示约0.08％的丙酮残留，酸碱摩尔比为1:2。DSC和TGA结果如图40所示。TGA结果显示在120-230℃之间有4.1％的失重，为水的脱去。DSC显示在85℃和155℃有两个宽的吸热峰，在230℃有另一个吸热峰，归为脱水和熔点。因此，柠檬酸盐晶型I可能是水合物。
柠檬酸盐晶型I的粉末X射线衍射图如图39所示，数据如表29所示：
表29
1.11苹果酸盐晶型I
室温下，约30mg的游离碱晶型I加到所选溶剂中。将苹果酸固体(1.1eq.)加到此悬浊液中。实验过程和结果如表30所示。
表30苹果酸盐的制备
苹果酸盐晶型I具有高结晶度的团聚的细小晶体。1H-NMR显示含有0.06％的丙酮残留，酸碱摩尔比为1:2。DSC和TGA结果如图42所示。TGA结果显示在60℃之前有0.2％的失重，归为水和丙酮的脱去。DSC显示在217℃处有个吸热峰，归为熔点。因此，苹果酸盐晶型I是无水晶型。
苹果酸盐晶型I的粉末X射线衍射图如图41所示，数据如表31所示：
表31

2、分析方法
2.1偏光显微镜分析(PLM)
采用ECLIPSE LV100POL(Nikon,JPN)偏光显微镜进行PLM分析。将少量的样品平铺至载玻片上，滴入香柏油将样品分散后，加上盖玻片。随后，将样品置于显微镜下用4-20倍物镜观察。
2.2 X-射线粉末衍射(XRPD)
使用X射线衍射仪对固体样品进行检测。将样品平铺于零背景单晶硅样品盘上，将样品轻压铺平后，根据表32中参数进行分析。
表32 XRPD测试参数
2.3热重分析(TGA)
样品的TGA数据使用TA仪器进行收集。将1-5mg样品置于已去皮的开口铝制样品盘中，按照表33中仪器参数对样品进行加热测试，并使用TRIOS软件对数据进行分析处理。
表33 TGA测试参数

2.4差示扫描量热分析(DSC)
样品的DSC数据使用TA仪器进行收集。将1-3mg样品置于已扎孔的DSC样品盘中，按照表34中的参数进行加热测定，并使用TRIOS软件对数据进行分析处理。
表34 DSC测试参数
2.5动态水蒸气吸附(DVS)
样品的水分吸脱附分析使用DVS仪器进行。将一定量的样品置于已去皮的样品盘中，样品重量自动称量，按照表35中的参数对样品进行分析。
表35无水晶型的DVS分析参数
2.6核磁共振质谱(1H-NMR)
1H-NMR采用布鲁克400MHz仪器采集数据。样品用DMSO-d6作为溶剂，测试参数如表36所示。并使用MestReNova软件对数据进行分析处理。
表36 1H-NMR测试参数
2.7高效液相色谱法(HPLC)
HPLC分析采用Agilent HPLC 1260系列仪器。溶解度和稳定性测试的HPLC方法如表37所示。
表37溶解度和稳定性测试的HPLC方法
3、式(A)所示化合物酸式盐、游离碱晶型评估
在本盐型筛选研究中，获得8个无水晶型：甲磺酸盐、苯磺酸盐、2个草酸盐、马来酸盐、磷酸盐、富马酸盐和苹果酸盐。
基于盐的固态表征数据和游离态的晶型筛选结果，选择富马酸盐晶型I、苹果酸盐晶型I和游离碱晶型III进行溶解度、稳定性研究以及药物代谢研究。
3.1溶解度研究
富马酸盐晶型I和苹果酸盐晶型I在37℃的生物媒介和水中进行溶解度测试。所有结果如表38所示。
富马酸盐晶型I和苹果酸盐晶型I的最大溶解度出现在0.5小时的SGF介质中(分别为0.21mg/mL和0.20mg/mL)，并且随着时间的延长溶解度降低。所有的晶型在水中几乎不溶(<LOQ)。
表38溶解度测试结果
LOQ:～1.0μg/mL
盐的溶解度用游离碱进行计算。
3.2固态稳定性研究
富马酸盐晶型I、苹果酸盐晶型I和游离碱晶型III的固态稳定性在60℃/闭口和40℃/75％RH条件下进行7天测试，实验结果如表39所示。
从表39可知，富马酸盐晶型I、苹果酸盐晶型I和游离碱晶型III的晶型稳定性良好，两种测试条件下三个化合物的晶型均未改变。富马酸盐晶型I在40℃/75％RH条件下，化学纯度稍有降低，但在闭口高温条件下稳定性也良好，储存时需控制湿度。其余两种晶型在两种条件下化学稳定性良好。
表39稳定性评估结果
3.3游离碱无定型的评估
各称取100mg本发明所得游离碱无定型样品加溶剂形成悬浊液，室温搅拌2天，结果如下：
由上述结果可知，游离碱无定型样品不稳定，极易在溶剂中转化为其他晶型。
3.4药物代谢研究
将富马酸盐晶型Ⅰ、苹果酸盐晶型Ⅰ、游离碱晶型Ⅰ及游离碱晶型Ⅲ，进行大鼠和犬的体内药物代谢实验。
将SD大鼠分别单次灌胃给予富马酸盐晶型Ⅰ、苹果酸盐晶型Ⅰ、游离碱晶型Ⅰ及游离碱晶型Ⅲ，于不同时间点采集血样，测定给予受试物后大鼠血浆中药物浓度并计算相关药代参数。
将犬分别单次灌胃给予富马酸盐晶型Ⅰ、游离碱晶型Ⅰ及游离碱晶型Ⅲ，于不同时间点采集血样，测定给予受试物后犬血浆中药物浓度并计算相关药代参数。
3.4.1、受试样品溶液制备
配制方法如下：
代号为富马酸盐晶型Ⅰ、苹果酸盐晶型Ⅰ、游离碱晶型Ⅰ及游离碱晶型Ⅲ的溶媒是0.5％MC+1％tween 80。具体配制方法为：取待供试品10mg，加0.5％MC和1％tween 80涡旋混合，得浓度为1mg/mL的溶液；使用前新鲜制备，涡旋混匀，备用。
3.4.2、供试品溶液分析
配制的供试品溶液由本实验机构分析部门采用LC-MS/MS进行检测分析.
3.4.3、动物接收与适应
健康雄性SD大鼠或犬用于该研究。所有动物给药前禁食，给药后4小时恢复给食。
3.4.4、样品采集与处理
采血时间点为：
口服给药：给药后0.25，0.5，1，2，4，6，8，12和24h。(≤2hr的时间点±2min，其余各点±15min。)
每只动物每次经颈静脉采集血液约0.1mL，以EDTA或肝素钠作为抗凝剂。血液样本采集后置于冰上，离心分离血浆(离心条件：10000转/分钟，5分钟，4℃)。
血浆样品由实验机构分析部门采用LC-MS/MS进行分析。
3.4.5、药物代谢动力学分析
根据药物的血药浓度数据，使用药代动力学计算软件WinNonlin非房室模型分别计算供试品的药代动力学参数AUC0→t、AUC0→∞、Cmax、Tmax和T1/2及其平均值和标准差。
3.4.6、实验结果
单次灌胃口服富马酸盐晶型Ⅰ、苹果酸盐晶型Ⅰ、游离碱晶型Ⅰ和游离碱晶型Ⅲ后血浆中的主要药代动力学参数如表40、表41所示。
表40大鼠药代动力学参数
表41犬药代动力学参数
上述药代结果可以看出，无论是大鼠还是犬的药代数据，富马酸盐晶型Ⅰ、苹果酸盐晶型Ⅰ、游离碱晶型Ⅲ的体内暴露量均高于游离碱晶型Ⅰ。游离碱晶型Ⅰ两个批次的游离碱晶型Ⅰ的PLM图见图45和图46，从图中可以看出，均为粒度很小的团聚物，在实际生产过程中过滤非常困难，不利于放大生产。
综上所述，富马酸盐晶型Ⅰ和苹果酸盐晶型Ⅰ的动物体内暴露量优异，固态稳定性良好，且适合放大生产，适合进一步进行研究开发。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (10)

  1. 一种式A化合物的晶型，其特征在于，所述晶型选自下组：游离碱晶型III、富马酸盐晶型I、苹果酸盐晶型I，
    其中，
    所述游离碱晶型III的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.8±0.2°、7.3±0.2°、8.2±0.2°、9.3±0.2°、10.4±0.2°、10.7±0.2°、13.0±0.2°、13.3±0.2°、14.7±0.2°、16.7±0.2°、18.4±0.2°、19.0±0.2°、23.4±0.2°；
    所述富马酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、8.7±0.2°、10.6±0.2°、14.6±0.2°、15.5±0.2°、17.5±0.2°、18.0±0.2°、19.2±0.2°、27.1±0.2°；
    所述苹果酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、9.3±0.2°、10.7±0.2°、11.2±0.2°、11.8±0.2°、15.3±0.2°、17.3±0.2°、17.6±0.2°、18.3±0.2°、19.0±0.2°、23.5±0.2°、25.7±0.2°、28.0±0.2°。
  2. 如权利要求1所述晶型，其特征在于，
    所述游离碱晶型III的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.8±0.2°、7.3±0.2°、7.8±0.2°、8.2±0.2°、9.3±0.2°、10.4±0.2°、10.7±0.2°、11.0±0.2°、13.0±0.2°、13.3±0.2°、14.0±0.2°、14.7±0.2°、15.2±0.2°、16.7±0.2°、17.5±0.2°、18.4±0.2°、19.0±0.2°、21.1±0.2°、22.2±0.2°、23.4±0.2°、29.5±0.2°；
    所述富马酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、8.7±0.2°、10.6±0.2°、11.0±0.2°、11.9±0.2°、12.1±0.2°、12.5±0.2°、14.6±0.2°、15.5±0.2°、15.9±0.2°、17.5±0.2°、18.0±0.2°、19.2±0.2°、26.7±0.2°、27.1±0.2°；
    所述苹果酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、9.3±0.2°、10.2±0.2°、10.7±0.2°、11.2±0.2°、11.8±0.2°、13.2±0.2°、15.3±0.2°、16.2±0.2°、17.3±0.2°、17.6±0.2°、18.3±0.2°、19.0±0.2°、23.5±0.2°、24.8±0.2°、25.7±0.2°、27.6±0.2°、28.0±0.2°。
  3. 如权利要求1所述晶型，其特征在于，
    所述游离碱晶型III的X射线粉末衍射图谱在如下2θ值处具有特征峰：5.1±0.2°、5.8±0.2°、7.3±0.2°、7.8±0.2°、8.2±0.2°、9.3±0.2°、10.4±0.2°、10.7±0.2°、11.0±0.2°、13.0±0.2°、13.3±0.2°、14.0±0.2°、14.7±0.2°、15.2±0.2°、16.7±0.2°、17.5±0.2°、18.4±0.2°、19.0±0.2°、21.1±0.2°、21.6±0.2°、22.2±0.2°、23.4±0.2°、24.9±0.2°、25.9±0.2°、26.3±0.2°、27.9±0.2°、28.4±0.2°、29.5±0.2°、30.0±0.2°、35.6±0.2°；
    所述富马酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、8.2±0.2°、8.7±0.2°、9.9±0.2°、10.6±0.2°、11.0±0.2°、11.9±0.2°、12.1±0.2°、12.5±0.2°、14.6±0.2°、15.5±0.2°、15.9±0.2°、16.5±0.2°、17.5±0.2°、18.0±0.2°、19.2±0.2°、21.2±0.2°、22.2±0.2°、23.9±0.2°、25.5±0.2°、26.7±0.2°、27.1±0.2°、28.8±0.2°；
    所述苹果酸盐晶型I的X射线粉末衍射图谱在如下2θ值处具有特征峰：6.0±0.2°、9.3±0.2°、10.2±0.2°、10.7±0.2°、11.2±0.2°、11.8±0.2°、12.1±0.2°、13.2±0.2°、15.3±0.2°、16.2±0.2°、16.8±0.2°、17.1±0.2°、17.3±0.2°、17.6±0.2°、18.3±0.2°、19.0±0.2°、23.5±0.2°、24.8±0.2°、25.2±0.2°、25.7±0.2°、27.6±0.2°、28.0±0.2°、29.7±0.2°。
  4. 如权利要求1所述晶型，其特征在于，
    所述游离碱晶型III的X射线粉末衍射图谱基本如图5所示；
    所述富马酸盐晶型I的X射线粉末衍射图谱基本如图37所示；
    所述苹果酸盐晶型I的X射线粉末衍射图谱基本如图41所示。
  5. 如权利要求1所述晶型，其特征在于，所述晶型选自下组：富马酸盐晶型I、苹果酸盐晶型I。
  6. 如权利要求1所述晶型，其特征在于，所述晶型为富马酸盐晶型I。
  7. 一种游离碱晶型III的制备方法，其特征在于，所述方法选自下组：
    方法一：将式A所示化合物与溶剂m混合，溶清，结晶析出；所述溶剂m选自正丁酮、丙酮和乙腈中的一种；
    方法二：将式A所示化合物与溶剂n混合，高温打浆，结晶析出；所述溶剂n选自乙酸乙酯、乙酸异丙酯和甲基叔丁基醚中的一种。
  8. 一种富马酸盐晶型I或苹果酸盐晶型I的制备方法，其特征在于，所述方法选自下组：
    方法一：将式A化合物溶解在有机溶剂中，室温搅拌，然后加入富马酸或苹果酸，继续搅拌，固液分离得到所述富马酸盐晶型I或苹果酸盐晶型I；
    方法二：向式A化合物的富马酸盐或苹果酸盐中加入有机溶剂，悬浮搅拌，固液分离得到所述富马酸盐晶型I或苹果酸盐晶型I。
  9. 一种权利要求1所述晶型的用途，其特征在于，用于制备药物，所述药物用于选自下组的用途：
    1)治疗与RET调节有关的疾病或病症；
    2)治疗肿瘤；
    3)抑制细胞增殖。
  10. 如权利要求9所述用途，其特征在于，所述肿瘤选自下组：甲状腺癌、非小细胞肺癌。
PCT/CN2023/095142 2023-05-18 2023-05-18 一种嘧啶并氮杂环类化合物的晶型及其制备方法及应用 Ceased WO2024234384A1 (zh)

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WO2020175968A1 (ko) * 2019-02-28 2020-09-03 주식회사 보로노이 N을 포함하는 헤테로아릴 유도체 및 이를 유효성분으로 포함하는 약학적 조성물
WO2020233641A1 (zh) * 2019-05-20 2020-11-26 浙江同源康医药股份有限公司 用作ret激酶抑制剂的化合物及其应用
CN113698390A (zh) 2020-05-20 2021-11-26 浙江同源康医药股份有限公司 用作ret激酶抑制剂的化合物及其应用

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WO2020175968A1 (ko) * 2019-02-28 2020-09-03 주식회사 보로노이 N을 포함하는 헤테로아릴 유도체 및 이를 유효성분으로 포함하는 약학적 조성물
WO2020233641A1 (zh) * 2019-05-20 2020-11-26 浙江同源康医药股份有限公司 用作ret激酶抑制剂的化合物及其应用
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