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WO2024199167A1 - 三芳香环类衍生物的盐、其晶型、制备方法和用途 - Google Patents

三芳香环类衍生物的盐、其晶型、制备方法和用途 Download PDF

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Publication number
WO2024199167A1
WO2024199167A1 PCT/CN2024/083481 CN2024083481W WO2024199167A1 WO 2024199167 A1 WO2024199167 A1 WO 2024199167A1 CN 2024083481 W CN2024083481 W CN 2024083481W WO 2024199167 A1 WO2024199167 A1 WO 2024199167A1
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Prior art keywords
formula
compound
salt
tartrate
hemifumarate
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PCT/CN2024/083481
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English (en)
French (fr)
Inventor
周文波
陈煌
鲁健
童孝田
彭世鸿
刘明耀
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Shanghai Yuyaobiotech Ltd
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Shanghai Yuyaobiotech Ltd
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Priority to CN202480018966.XA priority Critical patent/CN120981455A/zh
Publication of WO2024199167A1 publication Critical patent/WO2024199167A1/zh
Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings

Definitions

  • the present application relates to the field of pharmaceutical chemistry. Specifically, the present application relates to a triaromatic ring derivative, specifically a salt of (6-((5-(3-(4-(trifluoromethyl)phenyl)-1,2,4-oxadiazol-5-yl)pyrazin-2-yl)oxy)-1-methyl-1H-indol-2-yl)(4-(4-(2,2,2-trifluoroethoxy)benzyl)piperazin-1-yl)methanone, its crystal form and pharmaceutical composition, preparation method and use.
  • a triaromatic ring derivative specifically a salt of (6-((5-(3-(4-(trifluoromethyl)phenyl)-1,2,4-oxadiazol-5-yl)pyrazin-2-yl)oxy)-1-methyl-1H-indol-2-yl)(4-(4-(2,2,2-trifluoroethoxy)benzyl)piperazin-1-yl)me
  • Cide patent application CN112300145A discloses a triaromatic ring compound, whose chemical name is (6-((5-(3-(4-(trifluoromethyl)phenyl)-1,2,4-oxadiazol-5-yl)pyrazin-2-yl)oxy)-1-methyl-1H-indol-2-yl)(4-(4-(2,2,2-trifluoroethoxy)benzyl)piperazin-1-yl)methanone, and its structural formula is shown in formula (I):
  • the compound of formula (I) is an inhibitor targeting the bifunctional phosphorylation site of STAT3 and is used for preventing and/or treating diseases caused or regulated by the activation of the bifunctional phosphorylation site of STAT3, and has good clinical prospects.
  • salts For different active pharmaceutical molecules, especially oral solid drugs, different solid forms (salts, crystal forms) often affect the stability, solubility, purity, dissolution, and bioavailability of the drug, and only based on the molecular structure, it is usually impossible to accurately and reasonably predict whether it can be salified, whether it can be crystallized after salification, and how the properties after crystallization are.
  • some salts are solid at ambient temperature, while other salts are liquid or oily at ambient temperature.
  • some salt forms can remain stable under extreme conditions (such as heating), while other salts are easy to decompose under normal conditions (such as room temperature). Therefore, the development of salt forms for different active pharmaceutical molecules is an unpredictable process.
  • the salt of a compound may exist in one or more crystal forms, but its existence and characteristics cannot be specifically expected.
  • the present application provides salts of the compound of formula (I) and crystal forms of the salts.
  • the salts and crystal forms of the salts provided in the present application have at least one of the following advantages: good stability, high solubility, high bioavailability, good efficacy, good tablet dissolution, good tablet compressibility, good fluidity, good crystal morphology, good crystal size distribution, stable storage, simple and reliable preparation method, and great development value.
  • One aspect of the present application is to provide a salt of a compound of formula (I),
  • the salt is an acid salt, wherein the molar ratio of the compound of formula (I) to the acid is 0.5-2.
  • the salt of the compound of formula (I) is selected from fumarate, L-tartrate, hydrochloride, methanesulfonate and oxalate.
  • the salt of the compound of formula (I) is selected from fumarate.
  • the molar ratio of the compound of formula (I) to fumaric acid is 1:0.5-1; more preferably 1:0.5.
  • the salt of the compound of formula (I) is a hemifumarate, and the structural formula is shown in formula (II):
  • the hemifumarate salt of the compound of formula (I) is substantially in crystalline form or amorphous form; more preferably, it is substantially in crystalline form.
  • the hemifumarate salt of the compound of formula (I) may be in one or more crystalline forms; specifically, it may be at least one of an anhydrate, a hydrate or a solvate.
  • Another aspect of the present application is to provide a method for preparing the hemifumarate of the compound of formula (I), which comprises dissolving the compound of formula (I) in solvent 1, and then adding fumaric acid or a fumaric acid solution to react to obtain the hemifumarate.
  • the preparation method of the hemifumarate of the compound of formula (I) comprises the following steps:
  • Step A dissolving an appropriate amount of the compound of formula (I) in solvent 1;
  • Step B dissolving an appropriate amount of fumaric acid in an organic solvent
  • Step C combining the above two solutions, stirring to precipitate a solid or adding solvent 2 and stirring to precipitate a solid;
  • Step D Separate the solid to obtain the hemifumarate of the compound of formula (I).
  • the solvent 1 is selected from any one or a combination of alcohols, ethers, nitriles, esters and ketones; preferably, the alcohol is selected from any one or a combination of methanol, ethanol, n-propanol, sec-butanol, n-butanol and trifluoroethanol; the ether is selected from any one or a combination of diethyl ether and methyl tert-butyl ether; the nitriles are selected from acetonitrile; the esters are selected from any one or a combination of ethyl acetate, isopropyl acetate and n-butyl acetate; the ketones are selected from any one or a combination of acetone and butanone.
  • the alcohol is selected from any one or a combination of methanol, ethanol, n-propanol, sec-butanol, n-butanol and trifluoroethanol
  • the ether is selected from any one or
  • the solvent 1 is selected from any one of methanol, ethyl acetate, acetonitrile or a combination thereof.
  • the organic solvent is selected from alcohols.
  • the organic solvent is selected from methanol.
  • the solvent 2 is selected from any one of water, isopropyl ether, isopropyl alcohol, n-heptane and methylcyclohexane or a combination thereof.
  • the solvent 2 is selected from any one of water and isopropyl ether or a combination thereof.
  • the molar ratio of the compound of formula (I) to fumaric acid is 1:0.5-1.5; preferably, the molar ratio is 1:1.
  • the mass volume ratio (mg/ml) of the compound of formula (I) to solvent 1 is ⁇ 100:1; preferably ⁇ 80:1; more preferably ⁇ 50:1.
  • the mass volume ratio (mg/ml) of compound (I) to solvent 1 is 25:1.
  • the hemifumarate of the compound of formula (I) is in a crystalline form, and using Cu-K ⁇ radiation, the X-ray powder diffraction (XRPD) pattern of the hemifumarate crystalline form expressed in 2 ⁇ angles has a characteristic peak at at least one of 15.7° ⁇ 0.2°, 16.1° ⁇ 0.2° and 9.4° ⁇ 0.2°. Preferably, there are two or three characteristic peaks.
  • the XRPD spectrum of the hemi-fumarate crystalline form expressed in 2 ⁇ angle has characteristic peaks at 15.7° ⁇ 0.2°, 16.1° ⁇ 0.2° and 9.4° ⁇ 0.2°.
  • the XRPD spectrum of the hemi-fumarate crystalline form also has characteristic peaks at at least one of 14.0° ⁇ 0.2°, 17.1° ⁇ 0.2°, 17.9° ⁇ 0.2° and 19.2° ⁇ 0.2° 2 ⁇ .
  • the XRPD spectrum of the hemi-fumarate crystalline form also has characteristic peaks at at least one of 18.7° ⁇ 0.2°, 21.8° ⁇ 0.2°, 23.4° ⁇ 0.2 and 27.1° ⁇ 0.2°2 ⁇ .
  • the hemifumarate crystalline form is the hemifumarate Form 1 of the compound of formula (I) (referred to as hemifumarate Form 1).
  • the XRPD spectrum of the hemi-fumarate salt Form 1 also has characteristic peaks at at least one of 7.0° ⁇ 0.2°, 10.2° ⁇ 0.2°, 24.5° ⁇ 0.2° and 20.1° ⁇ 0.2°2 ⁇ .
  • the XRPD spectrum of Form 1 is 7.0° ⁇ 0.2°, 8.2° ⁇ 0.2°, 9.4° ⁇ 0.2°, 9.6° ⁇ 0.2°, 10.2° ⁇ 0.2°, 12.0° ⁇ 0.2°, 14.0° ⁇ 0.2°, 15.7° ⁇ 0.2°, 16.1° ⁇ 0.2°, 17.1° ⁇ 0.2°, 17.9° ⁇ 0.2°, 18.7° ⁇ 0.2°, 19.2° ⁇ 0.2°, 20.1° ⁇ 0.2°, 20.9° ⁇ 0.2°, 21.
  • the hemifumarate salt Form 1 has an XRPD pattern substantially the same as that of Figure 1.
  • the hemifumarate Form 1 has a TGA spectrum substantially the same as that of Figure 2.
  • the DSC spectrum of the hemi-fumarate Form 1 shows that its melting point is 156°C ⁇ 3°C.
  • the hemifumarate salt Form 1 has a DSC spectrum substantially the same as that of Figure 3.
  • the hemifumarate Form 1 has a DVS spectrum substantially the same as that of Figure 4.
  • the hemifumarate salt Form 1 is anhydrous.
  • the hemifumarate Form 1 has an XRD single crystal simulation pattern substantially as shown in Figure 20.
  • the crystalline form is hemifumarate Form 2 of the compound of formula (I) (referred to as hemifumarate Form 2).
  • the XRPD spectrum of the hemifumarate Form 2 also has characteristic peaks at at least one of 16.5° ⁇ 0.2°, 17.3° ⁇ 0.2°, 19.5° ⁇ 0.2°, 24.8° ⁇ 0.2° and 27.6° ⁇ 0.2°2 ⁇ .
  • the XRPD spectrum of the hemifumarate Form 2 is 4.2° ⁇ 0.2°, 5.0° ⁇ 0.2°, 9.3° ⁇ 0.2°, 9.9° ⁇ 0.2°, 11.2° ⁇ 0.2°, 12.6° ⁇ 0.2°, 14.0° ⁇ 0.2°, 15.1° ⁇ 0.2°, 15.6° ⁇ 0.2°, 16.2° ⁇ 0. 2°, 16.5° ⁇ 0.2°, 17.0° ⁇ 0.2°, 17.3° ⁇ 0.2°, 18.1° ⁇ 0.2°, 18.4° ⁇ 0.2°, 18.7° ⁇ 0.2°, 19.2° ⁇ 0.2°, 19.5° ⁇ 0.2°, 19.8° ⁇ 0.2°, 20.0° ⁇ 0.2°, 20.4° ⁇ 0.
  • the hemifumarate salt Form 2 has an XRPD pattern substantially the same as that of Figure 6.
  • the hemifumarate Form 2 has a TGA spectrum substantially the same as that of Figure 7.
  • the hemi-fumarate salt Form 2 has a DSC spectrum substantially the same as that of Figure 8.
  • the hemifumarate Form 2 has a DVS spectrum substantially the same as that of Figure 9.
  • the hemifumarate Form 2 is anhydrous.
  • Another aspect of the present application is to provide a method for preparing a hemifumarate of a compound of formula (I) in a crystalline form, the preparation method comprising any one of the following methods:
  • Step A dissolving an appropriate amount of the compound of formula (I) in solvent 3;
  • Step B dissolving an appropriate amount of fumaric acid in an organic solvent
  • Step C combining the above two solutions, stirring to precipitate or adding solvent 4 and stirring to precipitate to form a suspension;
  • Step D Continue stirring the suspension, and stir it under low temperature to crystallize, separate and dry to obtain the hemifumarate of the compound of formula (I) in crystalline form.
  • the solvent 3 is selected from one or a combination of alcohols, ethers, nitriles, esters and ketones; preferably, the alcohol is selected from any one or a combination of methanol, ethanol, n-propanol, sec-butanol, n-butanol and trifluoroethanol; the ether is selected from any one or a combination of diethyl ether and methyl tert-butyl ether; the nitrile is selected from acetonitrile; the ester is selected from any one or a combination of ethyl acetate, isopropyl acetate and n-butyl acetate; the ketone is selected from any one or a combination of acetone and butanone.
  • the alcohol is selected from any one or a combination of methanol, ethanol, n-propanol, sec-butanol, n-butanol and trifluoroethanol
  • the ether is selected from any one or a
  • the solvent 3 is selected from any one of methanol, ethyl acetate, acetonitrile or a mixture thereof.
  • the organic solvent is selected from alcohols.
  • the organic solvent is selected from methanol.
  • the solvent 4 is selected from one or a combination of water, isopropyl ether, isopropanol, n-heptane and methylcyclohexane.
  • the solvent 4 is selected from any one of water and isopropyl ether or a combination thereof.
  • the compound of formula (I) is heated and dissolved in solvent 3; the fumaric acid is heated and dissolved in an organic solvent; preferably, the temperature of the heating and dissolving is ⁇ 40°C; more preferably 50°C-60°C;
  • the stirring time in step C is ⁇ 1h.
  • the low temperature environment in step D is ⁇ 30°C; more preferably 0°C-20°C.
  • the low temperature environment in step D is 4°C.
  • the drying time is 8-24 hours; the drying temperature is 40°C.
  • the molar ratio of the compound of formula (I) to fumaric acid is 1:0.5-1.5.
  • the mass volume ratio (mg/ml) of the compound of formula (I) and the solvent 3 is ⁇ 100:1; preferably 80:1; more preferably 50:1.
  • the mass volume ratio (mg/ml) of compound (I) and solvent 3 is 25:1.
  • the solvent 5 is selected from one of alcohols, ketones, esters, ethers, dimethyl sulfoxide and tetrahydrofuran or a mixture thereof; more preferably, the solvent 5 is selected from tetrahydrofuran.
  • the solvent 6 is selected from one of water, n-heptane, isopropyl ether, dichloromethane, chloroform, toluene, methylcyclohexane or a mixture thereof; more preferably, the solvent 6 is selected from n-heptane.
  • the solvent 5 is selected from tetrahydrofuran, and the solvent 6 is selected from n-heptane.
  • the volume ratio of the solvent 6 to the solvent 5 is ⁇ 1:1; more preferably ⁇ 1.5:1.
  • the temperature of the heating dissolution is ⁇ 40°C; more preferably 50°C-60°C.
  • the drying time is ⁇ 4h; and the drying temperature is ⁇ 10°C.
  • the drying time is 8-24 hours; the drying temperature is 40°C.
  • the solvent 7 is selected from one of alcohols, ketones, esters, ethers, dimethyl sulfoxide and tetrahydrofuran or a mixture thereof; more preferably, the solvent 7 is selected from alcohols and ketones; most preferably, the alcohol is selected from one of ethanol and methanol or a combination thereof; the ketone is selected from acetone.
  • the solvent 8 is selected from one of water, n-heptane, isopropyl ether, dichloromethane, chloroform, toluene, methylcyclohexane or a mixture thereof; more preferably, it is one of water and n-heptane or a mixture thereof.
  • the crude product of the hemifumarate salt of the compound of formula (I) is dissolved in ethanol.
  • the compound of formula (I) is dissolved in a mixed solvent system of methanol and water, wherein the volume ratio of methanol to water is 1:0.1-0.2, preferably 1:0.15.
  • the compound of formula (I) is dissolved in a mixed solvent system of acetone and n-heptane, wherein the volume ratio of acetone to n-heptane is 1:0.5-1, preferably 1:0.8.
  • the temperature of the heating and dissolving is ⁇ 40°C; more preferably 50°C-60°C.
  • the temperature of the cooling and stirring is ⁇ 30°C; more preferably 0°C-20°C.
  • the drying time is ⁇ 4h; and the drying temperature is ⁇ 10°C.
  • the drying time is 8-24 hours; the drying temperature is 40°C.
  • the solvent 9 is selected from one or a combination of alcohols, alkanes, esters, nitriles, ethers, benzene rings, ketones and water; more preferably, the alcohol is selected from one or a combination of methanol, ethanol and isopropanol; the alkane is selected from one or a combination of dichloromethane, n-heptane and methylcyclohexane; the ester is selected from one or a combination of ethyl acetate and isopropyl acetate; the nitriles are selected from acetonitrile; the ethers are selected from one or a combination of diethyl ether and isopropyl ether; the benzene rings are selected from toluene; the ketones are selected from one or a combination of acetone and butanone.
  • the alcohol is selected from one or a combination of methanol, ethanol and isopropanol
  • the alkane is selected from one
  • the solvent 9 is selected from a single solvent system, such as one of methanol, water, isopropyl ether, dichloromethane, isopropanol, methyl tert-butyl ether, toluene, acetonitrile, and diethyl ether.
  • the solvent 9 is a mixed solvent system with a specific volume ratio, such as a mixed solvent system of methanol and water, with a volume ratio of 2:1; a mixed solvent system of ethanol and water, with a volume ratio of 3:1; a mixed solvent system of acetone and water, with a volume ratio of 1:2; a mixed solvent system of acetonitrile and water, with a volume ratio of 1:4; a saturated water mixed solvent system of ethyl acetate, a mixed solvent system of ethanol and isopropyl ether, with a volume ratio of 1:1; a mixed solvent system of isopropyl alcohol and n-heptane, with a volume ratio of 1:1; a mixed solvent system of butanone and methylcyclohexane, with a volume ratio of 1:5; isopropyl ether and ethyl acetate
  • the mixed solvent system is 5:1 in volume ratio; the mixed solvent system of ethyl acetate and dichloromethane is
  • the drying time is ⁇ 4h; and the drying temperature is ⁇ 10°C.
  • the drying time is 8-24 hours; the drying temperature is 40°C.
  • the mass volume ratio (mg/ml) of the crude product of the hemifumarate salt of the compound of formula (I) to the solvent 9 is ⁇ 150:1; more preferably ⁇ 120:1.
  • the mass volume ratio of the crude product of the hemifumarate salt of the compound of formula (I) and the solvent 9 is 100:1, 80:1, 60:1, 50:1, 25:1, or 20:1.
  • Another aspect of the present application is to provide a method for preparing hemifumarate Form 1, which comprises exposing the hemifumarate of the compound of formula (I) obtained in crystalline form by any of the above methods (1)-(4) to an environment with a relative humidity of ⁇ 75% for a sufficient period of time to obtain hemifumarate Form 1.
  • the exposure temperature is ⁇ 40°C.
  • the sufficient time refers to the time until the hemi-fumarate Form 1 is determined to be obtained after XRPD detection; in some embodiments, the sufficient time is generally more than 2 days.
  • Another aspect of the present application is to provide a method for preparing hemifumarate Form 2, which comprises exposing the hemifumarate of the compound of formula (I) obtained in crystalline form by any of the above methods (1)-(4) to an environment with a relative humidity of ⁇ 10% for a sufficient period of time to obtain hemifumarate Form 2.
  • the exposure temperature is ⁇ 30°C.
  • the sufficient time refers to the time until the hemi-fumarate Form 2 is determined to be obtained after XRPD detection; in some embodiments, the sufficient time is generally more than 2 days.
  • Another aspect of the present application is to provide a crystalline form of an ethanol solvate Form 3 of a hemifumarate of the compound of formula (I) (hereinafter referred to as hemifumarate Form 3).
  • the XRPD spectrum of the hemifumarate Form 3 expressed in 2 ⁇ angles has a characteristic peak at at least one of 6.8° ⁇ 0.2°, 9.4° ⁇ 0.2°, 15.7° ⁇ 0.2° and 15.9° ⁇ 0.2°; preferably, it has characteristic peaks at two, three or four locations.
  • the XRPD spectrum of the hemifumarate Form 3 also has characteristic peaks at at least one of 17.8° ⁇ 0.2°, 18.5° ⁇ 0.2°, 19.0° ⁇ 0.2° and 22.1° ⁇ 0.2°2 ⁇ .
  • the XRPD spectrum of the hemifumarate Form 3 also has characteristic peaks at at least one of 23.9° ⁇ 0.2°, 25.1° ⁇ 0.2°, 26.2° ⁇ 0.2° and 28.6° ⁇ 0.2°2 ⁇ .
  • the XRPD pattern of the Form 3 is at 4.1° ⁇ 0.2°, 6.8° ⁇ 0.2°, 8.0° ⁇ 0.2°, 9.4° ⁇ 0.2°, 10.0° ⁇ 0.2°, 11.2° ⁇ 0.2°, 13.0° ⁇ 0.2°, 13.6° ⁇ 0.2°, 14.5° ⁇ 0.2°, 15.7° ⁇ 0.2°, 16.0° ⁇ 0.2°, 16.5° ⁇ 0.2°, 17.4° ⁇ 0.2°, 17.8° ⁇ 0.2°, 18.5° ⁇ 0.2°, 19.0° ⁇ 0.2°, 20.0° ⁇ 0.2°, 20.4° ⁇ 0.2°, 20.6° ⁇ 0.2°, There are characteristic peaks at 0.2°, 22.1° ⁇ 0.2°, 22.9° ⁇ 0.2°, 23.6° ⁇ 0.2°, 23.9° ⁇ 0.2°, 24.2° ⁇ 0.2°, 24.5° ⁇ 0.2°, 25.1° ⁇ 0.2°, 26.2° ⁇ 0.2°, 27.4° ⁇ 0.2°, 27.7° ⁇ 0.2°, 28.6° ⁇ 0.2°, 29.2° ⁇ 0.2°, 31.0° ⁇ 0.2°, 31.6
  • the hemifumarate salt Form 3 has an XRPD spectrum substantially the same as that of Figure 11.
  • the hemifumarate Form 3 has a TGA spectrum substantially the same as that of Figure 12.
  • the hemifumarate Form 3 has a DSC spectrum substantially the same as that of Figure 13.
  • the hemifumarate salt Form 3 has a solid-state 1 H-NMR spectrum substantially the same as that of FIG. 14 .
  • the molar ratio of ethanol to the hemifumarate of the compound of formula (I) in the hemifumarate Form 3 is 1:1.
  • Another aspect of the present application is to provide a method for preparing hemifumarate Form 3, the preparation method comprising the following steps:
  • the crystalline form of the hemifumarate of the compound of formula (I) is diffused and crystallized in an ethanol atmosphere to obtain the ethanol solvate Form 3.
  • the diffusion crystallization time is ⁇ 1 day; more preferably ⁇ 5 days.
  • the diffusion crystallization time is 6 days.
  • hemifumarate Form 4 Another aspect of the present application is to provide a crystalline form of a hydrate Form 4 of the hemifumarate of the compound of formula (I) (hereinafter referred to as hemifumarate Form 4).
  • the XRPD spectrum of the hemifumarate Form 4 expressed in 2 ⁇ angles has a characteristic peak at at least one of 9.3° ⁇ 0.2°, 15.6° ⁇ 0.2°, 16.0° ⁇ 0.2°, and 21.9° ⁇ 0.2°; preferably, there are two, three, or four characteristic peaks.
  • the XRPD spectrum of the hemifumarate Form 4 also has characteristic peaks at at least one of 17.1° ⁇ 0.2°, 22.8° ⁇ 0.2°, 24.1° ⁇ 0.2° and 25.0° ⁇ 0.2°2 ⁇ .
  • the XRPD spectrum of the hemifumarate Form 4 also has characteristic peaks at at least one of 10.0° ⁇ 0.2°, 9.5° ⁇ 0.2°, 19.0° ⁇ 0.2° and 20.3° ⁇ 0.2°2 ⁇ .
  • the XRPD spectrum of the hemifumarate Form 4 is at 3.6° ⁇ 0.2°, 3.8° ⁇ 0.2°, 6.9° ⁇ 0.2°, 8.1° ⁇ 0.2°, 8.8° ⁇ 0.2°, 9.3° ⁇ 0.2°, 9.5° ⁇ 0.2°, 10.0° ⁇ 0.2°, 11.7° ⁇ 0.2°, 13.7° ⁇ 0.2°, 15.6° ⁇ 0.2°, 16.0° ⁇ 0.2°, 16.4° ⁇ 0.2°, 17.1° ⁇ 0.2°, 17.8° ⁇ 0.2°, 18.6° ⁇ 0.2°, 19.0° ⁇ 0.2°, 20.3° ⁇ 0.2°, 21.4° ⁇ 0.2°, 23.
  • the hemifumarate salt Form 4 has an XRPD pattern substantially the same as that of Figure 15.
  • the hemifumarate Form 4 has a TGA spectrum substantially the same as that of Figure 16.
  • the hemifumarate Form 4 has a DSC spectrum substantially the same as that of Figure 17.
  • the hemifumarate Form 4 has a DVS spectrum substantially the same as that of Figure 18.
  • the hemifumarate salt Form 4 has a solid-state 1 H-NMR spectrum substantially the same as that of FIG. 19 .
  • Another aspect of the present application is to provide a method for preparing hemifumarate Form 4, the preparation method comprising any one of the following methods:
  • the esters are selected from ethyl acetate; the alcohols are selected from isopropanol and sec-butyl alcohol or a mixture thereof; the ethers are selected from diethyl ether; the nitriles are selected from acetonitrile; and the ketones are selected from acetone.
  • the solvent 10 is ethyl acetate.
  • the solvent 10 is a mixed solvent system with a specific volume ratio, such as a mixed solvent system of acetone and isopropyl ether, with a volume ratio of 1:2; a mixed solvent system of isopropanol and acetonitrile, with a volume ratio of 5:2; a mixed solvent system of sec-butanol and ethyl acetate, with a volume ratio of 5:4.
  • a mixed solvent system of acetone and isopropyl ether with a volume ratio of 1:2
  • a mixed solvent system of isopropanol and acetonitrile with a volume ratio of 5:2
  • a mixed solvent system of sec-butanol and ethyl acetate with a volume ratio of 5:4.
  • the mass volume ratio (mg/ml) of the hemi-fumarate salt of the compound of formula (I) to the solvent 10 is ⁇ 150:1; more preferably ⁇ 120:1.
  • the mass volume ratio of the hemifumarate salt of the compound of formula (I) and the solvent 10 is 100:1, 70:1 or 50:1.
  • the solvent 11 is selected from one or a combination of ester and ketone solvents; more preferably, the ester is ethyl acetate; and the ketone is acetone.
  • the solvent 11 is ethyl acetate.
  • the solvent 11 is acetone.
  • the evaporation temperature is ⁇ 10°C.
  • the evaporation temperature is room temperature or 40°C.
  • the volume ratio of ethyl acetate to toluene is 5:1.
  • the temperature of the heating and dissolving is ⁇ 40°C; more preferably 50°C-60°C.
  • the temperature of the cooling and stirring is ⁇ 30°C; more preferably 0°C-20°C.
  • the drying time is ⁇ 4h; and the drying temperature is ⁇ 10°C.
  • the drying time is 8-24 hours; the drying temperature is 40°C.
  • the hemifumarate Form 1 of the present application is placed under high temperature (40°C and 60°C), high humidity (25°C/90% RH) and accelerated (40°C/75% RH) conditions for 30 days, and the crystal form remains basically unchanged, and the increase of related substances is less than 0.01%, which has good crystal stability and chemical stability; the hemifumarate Form 2 is placed under high temperature (open, 60°C, ⁇ 30% RH) conditions for 10 days, and the crystal form remains basically unchanged, and the increase of related substances is less than 0.05%, which has good crystal stability and chemical stability.
  • the hemifumarate Form 1 of the present application has good blood concentration and curve area and excellent oral absolute bioavailability in SD rats.
  • its blood concentration and curve area are 1.9 times and 1.8 times that of the free state of the compound of formula (I), respectively, and its oral absolute availability is 3.2 times that of the free state of the compound of formula (I); the oral absolute availability of the hemifumarate Form 2 is 2.7 times that of the free state of the compound of formula (I).
  • the hemi-fumarate Form 1 of the present application can significantly inhibit the growth of human breast cancer MDA-MB-231 cell Balb/c nude mouse xenograft tumors, the tumor volume inhibition rate TGI (%) is 75.46%, and the relative tumor proliferation rate T/C after administration is 23.91%; the hemi-fumarate Form 1 of the present application can significantly inhibit the growth of human leukemia MOLM16 cell CB-17SCID mouse xenograft tumors and significantly inhibit the growth of human leukemia MOLM16 cell CB-17SCID mouse xenograft tumors.
  • the tablets have good dissolution. Compared with the oxalate Form 2, the tablets with the hemi-fumarate Form 1 and hemi-fumarate Form 2 as active ingredients of the present application have better in vitro dissolution.
  • the salt of the compound of formula (I) is selected from L-tartrate.
  • the molar ratio of the compound of formula (I) to L-tartaric acid is 1:0.5-2; more preferably 1:1.
  • the salt of the compound of formula (I) is selected from L-tartrate, and the structural formula is shown in formula (III):
  • the L-tartrate salt of the compound of formula (I) is substantially in crystalline form or amorphous form; more preferably, it is substantially in crystalline form.
  • the crystalline form of the L-tartrate of the compound of formula (I) may be one or more; specifically, it may be at least one of an anhydrate, a hydrate or a solvate.
  • Another aspect of the present application is to provide a method for preparing L-tartrate of the compound of formula (I), which comprises dissolving the compound of formula (I) in a solvent 12, and then adding L-tartaric acid or an L-tartaric acid solution to react.
  • the preparation method of the L-tartrate of the compound of formula (I) comprises the following steps:
  • Step A dissolving an appropriate amount of the compound of formula (I) in solvent 12;
  • Step B dissolving an appropriate amount of L-tartaric acid in solvent 13;
  • Step C combining the above two solutions, stirring to precipitate solids to form a suspension
  • Step D separating the solid in the suspension to obtain L-tartrate of the compound of formula (I);
  • the solvent 12 is selected from any one or a combination of benzene rings, alkanes, esters, alcohols, ethers, ketones, halogens, tetrahydrofuran, 1,4-dioxane and dimethyl sulfoxide; more preferably, the benzene rings are selected from toluene; the alkanes are selected from dichloromethane; the alcohols are selected from any one or a combination of trifluoroethanol, methanol, ethanol, n-propanol, sec-butanol and n-butanol; the ethers are selected from any one or a combination of diethyl ether and methyl tert-butyl ether; the esters are selected from any one or a combination of ethyl acetate, isopropyl acetate and n-butyl acetate; the ketones are selected from any one or a combination of acetone and butanone; the halogens
  • the solvent 12 is selected from any one of toluene, dichloromethane, trifluoroethanol, ether, ethyl acetate, isopropyl acetate, acetone, butanone, chloroform, tetrahydrofuran and 1,4-dioxane, or a combination thereof.
  • the solvent 12 is selected from toluene.
  • the solvent 13 is selected from any one of alcohols, alkanes, ketones and tetrahydrofuran or a combination thereof; more preferably, the alcohol is selected from ethanol; the alkane is selected from dichloromethane; and the ketone is selected from acetone.
  • the solvent 13 is selected from ethanol or tetrahydrofuran.
  • the solvent 13 is selected from a mixed solvent of acetone/ethanol or a mixed solvent of dichloromethane/ethanol.
  • the solvent 14 may be added to the suspension in step D before solid separation.
  • the solvent 14 is selected from any one of isopropyl ether, n-heptane, water, isopropanol, and methylcyclohexane, or a combination thereof.
  • the solvent 14 is selected from any one of isopropyl ether and n-heptane or a combination thereof.
  • the molar ratio of the compound of formula (I) to L-tartaric acid is 1:1-2.5; more preferably, the molar ratio is 1:1.
  • the mass volume ratio (mg/ml) of the compound of formula (I) to the solvent 12 is ⁇ 300:1; more preferably ⁇ 200:1; most preferably ⁇ 150:1.
  • the mass volume ratio (mg/ml) of the compound of formula (I) and solvent 12 is 100:1.
  • L-tartrate Form 1 a crystalline form of Form 1 of L-tartrate of the compound of formula (I) (hereinafter referred to as L-tartrate Form 1).
  • L-tartrate Form 1 Using Cu-K ⁇ radiation, the XRPD spectrum of the L-tartrate Form 1 expressed in 2 ⁇ angles has a characteristic peak at at least one of 7.8° ⁇ 0.2°, 14.6° ⁇ 0.2°, 18.3° ⁇ 0.2° and 23.6° ⁇ 0.2°; preferably, there are two, three or four characteristic peaks.
  • the XRPD spectrum of the L-tartrate salt Form 1 also has characteristic peaks at at least one of 20.0° ⁇ 0.2°, 14.1° ⁇ 0.2°, 13.5° ⁇ 0.2° and 16.1° ⁇ 0.2°2 ⁇ .
  • the XRPD spectrum of the L-tartrate salt Form 1 also has characteristic peaks at at least one of 19.8° ⁇ 0.2°, 13.3° ⁇ 0.2°, 17.0° ⁇ 0.2° and 15.3° ⁇ 0.2°2 ⁇ .
  • the XRPD pattern of the L-tartrate salt Form 1 is at 3.9° ⁇ 0.2°, 6.1° ⁇ 0.2°, 7.8° ⁇ 0.2°, 10.2° ⁇ 0.2°, 11.7° ⁇ 0.2°, 13.3° ⁇ 0.2°, 13.5° ⁇ 0.2°, 14.1° ⁇ 0.2°, 14.6° ⁇ 0.2°, 15.3° ⁇ 0.2°, 16.1° ⁇ 0.2°, 17.0° ⁇ 0.2°, 17.7° ⁇ 0.2°, 18.3° ⁇ 0.2°, 18.6° ⁇ 0.2°, 18.8° ⁇ 0.2°, 19.1° ⁇ 0.
  • the L-tartrate salt Form 1 has an XRPD pattern substantially the same as that of Figure 21.
  • the L-tartrate salt Form 1 has a TGA spectrum substantially the same as that of Figure 22.
  • the L-tartrate salt Form 1 has a DSC spectrum substantially the same as that of Figure 23.
  • the L-tartrate salt Form 1 has a DVS spectrum substantially the same as that of Figure 24.
  • the L-tartrate salt Form 1 is a hydrate.
  • Another aspect of the present application is to provide a method for preparing L-tartrate Form 1, the preparation method comprising any one of the following methods:
  • Step A dissolving an appropriate amount of the compound of formula (I) in a solvent 15;
  • Step B dissolving an appropriate amount of L-tartaric acid in solvent 16;
  • Step C combining the above two solutions, stirring to precipitate solids to form a suspension
  • Step D Continue stirring or add solvent 17 to the suspension and continue stirring, crystallize, centrifuge, and dry to obtain L-tartrate salt Form 1.
  • the solvent 15 is selected from benzene rings; preferably, the benzene rings are selected from toluene.
  • the solvent 15 is selected from toluene.
  • the solvent 16 is selected from alcohols; more preferably, the alcohol is selected from ethanol.
  • the solvent 16 is selected from ethanol.
  • the solvent 17 is selected from isopropyl ether.
  • the solution merging in step C is performed by dropwise adding the solution of step B into the solution of step A.
  • the drying time in step D is ⁇ 5 h; preferably ⁇ 10 h.
  • the drying time is 12-24 hours.
  • the drying time is 16 hours.
  • the drying temperature is room temperature.
  • the stirring temperature in step D is low temperature, which is ⁇ 30°C; more preferably 0°C-20°C;
  • the stirring temperature in step D is 4°C.
  • the molar ratio of the compound of formula (I) to L-tartaric acid is 1:1-2.5; more preferably, the molar ratio is 1:1.
  • the mass volume ratio (mg/ml) of the compound of formula (I) and the solvent 15 is ⁇ 300:1; preferably ⁇ 200:1; and more preferably ⁇ 150:1.
  • the mass volume ratio (mg/ml) of the compound of formula (I) and solvent 15 is 100:1.
  • the solvent 18 is selected from any one or a combination of esters, alkanes, alcohols, ethers and water; the esters are selected from isopropyl acetate; the alkanes are selected from any one or a combination of dichloromethane, n-heptane, n-hexane and methylcyclohexane; the alcohols are selected from any one or a combination of methanol and ethanol; The ethers are selected from methyl tert-butyl ether.
  • the solvent 18 is a mixed solvent system with a specific volume ratio, such as a mixed solvent of isopropyl acetate and n-heptane, with a volume ratio of 1:4; or a mixed solvent of dichloromethane and n-hexane, with a volume ratio of 1:4; or a mixed solvent of ethanol and n-heptane, with a volume ratio of 1:4; or a mixed solvent of methanol and water, with a volume ratio of 1:2; or a mixed solvent of dichloromethane and methylcyclohexane, with a volume ratio of 1:6; or a mixed solvent of methyl tert-butyl ether and ethanol, with a volume ratio of 4:1; or a mixed solvent of isopropyl acetate and n-hexane, with a volume ratio of 1:6.
  • a specific volume ratio such as a mixed solvent of isopropyl acetate and n-heptane, with a
  • the stirring temperature is low temperature, room temperature or high temperature.
  • the low temperature is ⁇ 30°C; more preferably 0°C-20°C.
  • the low temperature is 4°C-8°C.
  • the high temperature is ⁇ 30°C; more preferably 40°C-60°C.
  • the elevated temperature is 50°C.
  • the mass volume ratio (mg/mL) of the L-tartrate salt of the compound of formula (I) and the solvent 18 is 40:1.
  • L-tartrate Form 2 a crystalline form of Form 2 of L-tartrate of the compound of formula (I) (hereinafter referred to as L-tartrate Form 2), using Cu-K ⁇ radiation, and the L-tartrate Form 2 has an XRPD spectrum substantially the same as that of Figure 26.
  • the L-tartrate salt Form 2 has a TGA spectrum substantially the same as that of Figure 27.
  • the L-tartrate salt Form 2 has a DSC spectrum substantially the same as that of Figure 28.
  • the L-tartrate salt Form 2 is a hydrate.
  • Another aspect of the present application is to provide a method for preparing L-tartrate Form 2, which comprises exposing L-tartrate Form 1 to an environment with a relative humidity ⁇ 75% for a sufficient period of time to obtain L-tartrate Form 2.
  • the exposure temperature is ⁇ 40°C.
  • the relative humidity is 75% or 97%; and the exposure temperature is room temperature.
  • the sufficient time refers to the time until L-tartrate Form 2 is determined to be obtained after XRPD detection.
  • the sufficient period of time is 7 days.
  • L-tartrate Form 3 Another aspect of the present application is to provide a crystalline form of Form 3 of the L-tartrate salt of the compound of formula (I) (hereinafter referred to as L-tartrate Form 3), using Cu-K ⁇ radiation, and the L-tartrate Form 3 has an XRPD spectrum substantially the same as that of Figure 30.
  • the L-tartrate salt Form 3 has a TGA spectrum substantially the same as that of Figure 31.
  • the L-tartrate salt Form 3 has a DSC spectrum substantially the same as that of Figure 32.
  • the L-tartrate salt Form 3 is a hydrate.
  • Another aspect of the present application is to provide a method for preparing L-tartrate Form 3, the preparation method comprising:
  • L-tartrate salt Form 1 is suspended in solvent 19, stirred at low temperature to precipitate, and the crystals are separated to obtain L-tartrate salt Form 3; wherein solvent 19 is a mixed solvent of dimethyltetrahydrofuran and an alkane solvent.
  • the solvent 19 is a mixed solvent of dimethyltetrahydrofuran and methylcyclohexane, with a volume ratio of 1:4.
  • the temperature of the low-temperature stirring is ⁇ 30°C; preferably 0°C-20°C.
  • the low temperature stirring temperature is 4°C-8°C.
  • the stirring time is ⁇ 0.5 days; more preferably 1-10 days.
  • the stirring time is 3 days.
  • the mass volume ratio (mg/mL) of the L-tartrate salt Form 1 and the solvent 19 is 40:1.
  • L-tartrate Form 4 Another aspect of the present application is to provide a crystalline form of Form 4 of L-tartrate of the compound of formula (I) (hereinafter referred to as L-tartrate Form 4).
  • L-tartrate Form 4 Using Cu-K ⁇ radiation, the XRPD spectrum of the L-tartrate Form 4 expressed in 2 ⁇ angle has a characteristic peak at at least one of 3.6° ⁇ 0.2°, 11.0° ⁇ 0.2° and 14.3° ⁇ 0.2°; preferably, there are two or three characteristic peaks.
  • the XRPD spectrum of the L-tartrate salt Form 4 also has characteristic peaks at at least one of 6.6° ⁇ 0.2°, 7.3° ⁇ 0.2°, 8.4° ⁇ 0.2°, 17.7° ⁇ 0.2° and 18.0° ⁇ 0.2°2 ⁇ .
  • the XRPD spectrum of the L-tartrate salt Form 4 also has characteristic peaks at at least one of 13.2° ⁇ 0.2°, 13.8° ⁇ 0.2°, 18.5° ⁇ 0.2°19.9° ⁇ 0.2° and 21.8° ⁇ 0.2°2 ⁇ .
  • the XRPD pattern of the L-tartrate salt Form 4 is at 3.6° ⁇ 0.2°, 6.6° ⁇ 0.2°, 7.3° ⁇ 0.2°, 8.4° ⁇ 0.2°, 11.0° ⁇ 0.2°, 12.5° ⁇ 0.2°, 13.2° ⁇ 0.2°, 13.8° ⁇ 0.2°, 14.0° ⁇ 0.2°, 14. 3° ⁇ 0.2°, 14.7° ⁇ 0.2°, 15.6° ⁇ 0.2°, 16.0° ⁇ 0.2°, 16.9° ⁇ 0.2°, 17.5° ⁇ 0.2°, 17.7° ⁇ 0.2°, 18.0° ⁇ 0.2°, 18.5° ⁇ 0.2°, 18.8° ⁇ 0.2°, 19.2° ⁇ 0.2°, 19. 5° ⁇ 0.
  • the L-tartrate salt Form 4 has an XRPD pattern substantially the same as that of Figure 34.
  • the L-tartrate salt Form 4 has a TGA spectrum substantially the same as that of Figure 35.
  • the L-tartrate salt Form 4 has a DSC spectrum substantially the same as that of Figure 36.
  • the L-tartrate salt Form 4 is an anhydrate or a channel hydrate.
  • Another aspect of the present application is to provide a method for preparing L-tartrate Form 4, the preparation method comprising any one of the following methods:
  • the solvent 20 is selected from any one of ketones, esters, ethers and benzene rings or a mixture thereof; more preferably, the ketones are selected from any one of acetone and methyl isobutyl ketone or a combination thereof; the esters are selected from any one of ethyl acetate and isopropyl acetate or a combination thereof; the ethers are selected from methyl tert-butyl ether; and the benzene rings are selected from toluene.
  • the solvent 20 is a single solvent system, such as isopropyl acetate.
  • the solvent 20 is a mixed solvent system with a specific volume ratio, such as a mixed solvent of acetone and methyl tert-butyl ether, with a volume ratio of 1:1; or a mixed solvent system of ethyl acetate and toluene, with a volume ratio of 2:1.
  • the volatilization time is ⁇ 0.5 days; in some embodiments, the volatilization time is 1 day.
  • the slow volatilization is small pore volatilization.
  • the slow volatilization temperature is room temperature.
  • the solvent 21 is any one of ketones, alkanes, esters and ethers or a combination thereof; more preferably, the ketone is selected from any one of acetone and methyl isobutyl ketone or a combination thereof; the alkane is selected from n-heptane; the ester is selected from ethyl acetate; and the ether is selected from methyl tert-butyl ether.
  • the solvent 21 is a mixed solvent system with a specific volume ratio, such as a mixed solvent of acetone and n-heptane, with a volume ratio of 1:6; or a mixed solvent of ethyl acetate and n-heptane, with a volume ratio of 1:6; or a mixed solvent of isopropyl acetate and n-heptane, with a volume ratio of 1:4, or a mixed solvent of acetone and methyl tert-butyl ether, with a volume ratio of 1:4.
  • a specific volume ratio such as a mixed solvent of acetone and n-heptane, with a volume ratio of 1:6; or a mixed solvent of ethyl acetate and n-heptane, with a volume ratio of 1:6; or a mixed solvent of isopropyl acetate and n-heptane, with a volume ratio of 1:4, or a mixed solvent of acetone and methyl
  • the mass volume ratio (mg/mL) of the L-tartrate salt Form 1 and the solvent 21 is 40:1.
  • the solvent 22 is selected from any one or a combination of ketones, alkanes, esters and ethers; more preferably, the ketones are selected from any one or a combination of acetone and methyl isobutyl ketone; the alkanes are selected from n-heptane; the esters are selected from ethyl acetate; and the ethers are selected from methyl tert-butyl ether.
  • the solvent 22 is a mixed solvent system with a specific volume ratio, such as a mixed solvent of acetone and n-heptane, with a volume ratio of 1:6.
  • the molar ratio of the compound of formula (I) to L-tartaric acid is 1:1.
  • the mass volume ratio (mg/mL) of the compound of formula (I) and solvent 22 is 30:1.
  • the solvent 23 is selected from any one or a combination of alkanes, ketones and ethers; more preferably, the alkane is selected from n-heptane; the ketone is selected from butanone; and the ether is selected from methyl tert-butyl ether.
  • the solvent 23 is methyl tert-butyl ether.
  • the solvent 23 is a mixed solvent system with a specific volume ratio, such as a mixed solvent of butanone and n-heptane, with a volume ratio of 1:6.
  • the stirring temperature is low temperature or room temperature.
  • the low temperature is ⁇ 30°C; more preferably 0°C-20°C.
  • the low temperature is 4°C-8°C.
  • the mass volume ratio (mg/mL) of the L-tartrate salt of the compound of formula (I) and the solvent 23 is 40:1.
  • L-tartrate Form 5 Another aspect of the present application is to provide a crystalline form of Form 5 of L-tartrate of the compound of formula (I) (hereinafter referred to as L-tartrate Form 5), using Cu-K ⁇ radiation, the L-tartrate Form 5 has an XRPD pattern substantially the same as that of Figure 38.
  • Another aspect of the present application is to provide a method for preparing L-tartrate Form 5, which comprises forming a suspension of L-tartrate Form 1 in a mixed solvent of isopropanol and methyl tert-butyl ether, stirring to precipitate, separating the crystals, and obtaining L-tartrate Form 5.
  • the volume ratio of isopropanol to methyl tert-butyl ether is 1:1.
  • the stirring is stirring at room temperature.
  • the stirring time is 3 days.
  • the mass volume ratio (mg/mL) of the L-tartrate salt Form 1 and the mixed solvent is 40:1.
  • L-tartrate Form 6 Another aspect of the present application is to provide a crystalline form of L-tartrate Form 6 of the compound of formula (I) (hereinafter referred to as L-tartrate Form 6), using Cu-K ⁇ radiation, and the L-tartrate Form 6 has an XRPD spectrum substantially the same as that of Figure 39.
  • Another aspect of the present application is to provide a method for preparing L-tartrate Form 6, which comprises heating L-tartrate Form 1 to 70°C and maintaining the temperature for 10 minutes to obtain L-tartrate Form 6.
  • L-tartrate amorphous an amorphous form of L-tartrate of the compound of formula (I) (hereinafter referred to as L-tartrate amorphous), wherein the L-tartrate amorphous has no sharp diffraction peaks in the XRPD spectrum represented by 2 ⁇ using Cu-K ⁇ radiation.
  • the L-tartrate amorphous form has an XRPD pattern substantially the same as 40.
  • the L-tartrate amorphous form has a TGA pattern substantially the same as that of 41.
  • the L-tartrate amorphous form has a DSC spectrum substantially the same as that of 42.
  • the L-tartrate amorphous form has a DVS pattern substantially the same as that of Figure 43.
  • Another aspect of the present application is to provide a method for preparing amorphous L-tartrate, which comprises dissolving L-tartrate of the compound of formula (I) in dichloromethane, filtering, and concentrating the filtrate at high temperature to obtain amorphous L-tartrate.
  • the concentration is concentration under reduced pressure.
  • the high temperature is 40-70°C.
  • the elevated temperature is 60°C.
  • the mass volume ratio (mg/ml) of the L-tartrate salt of the compound of formula (I) and dichloromethane is 10:1.
  • the L-tartrate Form 1 of the present application was placed under long-term (25°C/60%RH) and accelerated (40°C/75%RH) conditions for 1 week, and the crystal form and purity remained basically unchanged. It was placed under high temperature (60°C, ⁇ 30%RH) conditions for 10 days, and its crystal form remained basically unchanged, and the increase of related substances was ⁇ 0.05%, which had good crystal stability and chemical stability; the L-tartrate Form 4 of the present application was placed under long-term (25°C/60%RH) and accelerated (40°C/75%RH) conditions for 1 week, and the crystal form and purity remained basically unchanged. It was placed under 40°C, 50°C and 55°C conditions for 30 days, and its crystal form remained basically unchanged, which had good crystal stability and chemical stability.
  • the L-tartrate salt Form 1 of the present application has good blood drug concentration and curve area and excellent oral absolute bioavailability in SD rats.
  • its blood drug concentration and curve area are 1.5 times and 1.8 times that of the free state of the compound of formula (I), respectively, and its oral absolute availability is 2.6 times that of the free state of the compound of formula (I);
  • the L-tartrate salt Form 4 of the present application has good blood drug concentration and curve area and excellent oral absolute bioavailability in SD rats.
  • the L-tartrate Form 1 of the present application has a significantly better tumor inhibition effect than the solvent in the NOD-SCID immunodeficient mouse Mol m16 subcutaneous tumor model, and at the same dose, its tumor inhibition effect is better than the free state of the compound of formula (I);
  • the L-tartrate Form 4 of the present application can significantly inhibit the growth of human breast cancer MDA-MB-231 cell Balb/c nude mouse xenograft tumors, the tumor volume inhibition rate TGI (%) is 79.43%, and the relative tumor proliferation rate T/C after administration is 20.37%;
  • the L-tartrate Form 4 of the present application significantly inhibits the growth of human breast cancer MDA-MB-231 cell Balb/c nude mouse xenograft tumors and the growth of human non-small cell lung cancer NCI-H460 cell Balb/c nude mouse xenograft tumors.
  • L-tartrate Form 4 and L-tartrate Form 1 of the present application have good fluidity and are both better than the oxalate Form 2.
  • the salt of the compound of formula (I) is selected from hydrochloride, and the structural formula is shown in formula (IV):
  • the hydrochloride salt of the compound of formula (I) is substantially in crystalline form or amorphous form; more preferably, it is substantially in crystalline form.
  • the hydrochloride of the compound of formula (I) may be in one or more crystalline forms; specifically, it may be at least one of an anhydrate, a hydrate or a solvate.
  • hydrochloride salt Form 1 Another aspect of the present application is to provide a crystalline form of a hydrochloride salt of the compound of formula (I) Form 1 (hereinafter referred to as hydrochloride salt Form 1).
  • hydrochloride salt Form 1 Using Cu-K ⁇ radiation, the XRPD spectrum of the hydrochloride salt Form 1 expressed in 2 ⁇ angles has a characteristic peak at at least one of 13.5° ⁇ 0.2°, 15.4° ⁇ 0.2°, 15.7° ⁇ 0.2°, 32.6° ⁇ 0.2° and 35.3° ⁇ 0.2°; preferably, there are two, three, four or five characteristic peaks.
  • the hydrochloride salt Form 1 has an XRPD pattern substantially the same as that of Figure 45.
  • the hydrochloride salt Form 1 has a TGA spectrum substantially the same as that of Figure 46.
  • the hydrochloride salt Form 1 has a DSC spectrum substantially the same as that of Figure 47.
  • Another aspect of the present application is to provide a method for preparing hydrochloride Form 1, the preparation method comprising the following steps:
  • Step A dissolving an appropriate amount of the compound of formula (I) in an ester solvent
  • Step B dissolving an appropriate amount of hydrochloric acid in an alcohol solvent
  • Step C combining the two solutions, stirring to precipitate solids to form a suspension
  • Step D Stir the suspension to crystallize, centrifuge and dry to obtain hydrochloride Form 1.
  • the ester solvent is selected from butyl acetate.
  • the alcohol solvent is selected from methanol.
  • the stirring temperature in step D is 4°C.
  • the drying time is 16 hours.
  • the drying temperature is room temperature.
  • the molar ratio of the compound of formula (I) to hydrochloric acid is 1:1.2.
  • the mass volume ratio (mg/ml) of the compound of formula (I) and the ester solvent is 100:1.
  • the salt of the compound of formula (I) is selected from methanesulfonate, and the structural formula is shown in formula (V):
  • the methanesulfonate salt of the compound of formula (I) is substantially in crystalline form or amorphous form; more preferably, it is substantially in crystalline form.
  • the crystalline form of the methanesulfonate salt of the compound of formula (I) may be one or more; specifically, it may be at least one of an anhydrate, a hydrate or a solvate.
  • mesylate salt Form 1 a crystalline form of Form 1 of the mesylate salt of the compound of formula (I) (hereinafter referred to as mesylate salt Form 1), using Cu-K ⁇ radiation, and the mesylate salt Form 1 has an XRPD spectrum substantially the same as that of Figure 49.
  • methanesulfonate Form 2 Another aspect of the present application is to provide a crystalline form of Form 2 of the methanesulfonate of the compound of formula (I) (hereinafter referred to as methanesulfonate Form 2).
  • methanesulfonate Form 2 Using Cu-K ⁇ radiation, the XRPD spectrum of the methanesulfonate Form 2 expressed in 2 ⁇ angles has a characteristic peak at at least one of 6.5° ⁇ 0.2°, 13.8° ⁇ 0.2°, 15.2° ⁇ 0.2° and 17.0° ⁇ 0.2°; preferably, there are characteristic peaks at two, three or four locations.
  • the XRPD spectrum of the mesylate salt Form 2 is also at 3.9° ⁇ 0.2°, 4.6° ⁇ 0.2°, 4.9° ⁇ 0.2°, 10.5° ⁇ 0.2°, 10.7° ⁇ 0.2°, 12.8° ⁇ 0.2°, 13.0° ⁇ 0.2°, 16.3° ⁇ 0.2°, 16.7° ⁇ 0.2°, 17.4° ⁇ 0.2°, 18.3° ⁇ 0.2°, There is a characteristic peak at least one of 18.8° ⁇ 0.2°, 19.6° ⁇ 0.2°, 20.6° ⁇ 0.2°, 21.2° ⁇ 0.2°, 22.4° ⁇ 0.2°, 23.0° ⁇ 0.2°, 23.9° ⁇ 0.2°, 25.6° ⁇ 0.2°, 25.8° ⁇ 0.2°, 26.6° ⁇ 0.2°, 29.6° ⁇ 0.2° and 31.4° ⁇ 0.2°2 ⁇ .
  • the mesylate salt Form 2 has an XRPD spectrum substantially the same as that of Figure 50.
  • the mesylate salt Form 2 has a TGA spectrum substantially the same as that of Figure 51.
  • the mesylate salt Form 2 has a DSC spectrum substantially the same as that of Figure 52.
  • Another aspect of the present application is to provide a method for preparing mesylate Form 2, the preparation method comprising:
  • the benzene ring solvent is toluene.
  • the mass volume ratio (mg/ml) of methanesulfonic acid and toluene is 72.5:1.
  • the mass volume ratio (mg/ml) of the compound of formula (I) and toluene is 100:1.
  • the molar ratio of the compound of formula (I) to methanesulfonic acid is 1:1.1.
  • the ether solvent is isopropyl ether.
  • the low temperature stirring temperature is 4°C.
  • mesylate salt Form 3 Another aspect of the present application is to provide a crystalline form of Form 3 of the mesylate salt of the compound of formula (I) (hereinafter referred to as mesylate salt Form 3), using Cu-K ⁇ radiation, and the mesylate salt Form 3 has an XRPD spectrum substantially the same as that of Figure 54.
  • the salt of the compound of formula (I) is selected from oxalate, and the structural formula is shown in formula (VI):
  • the oxalate salt of the compound of formula (I) is substantially present in crystalline or amorphous form.
  • the crystalline form of the oxalate of the compound of formula (I) may be one or more; specifically, it may be at least one of an anhydrate, a hydrate or a solvate.
  • Another aspect of the present application is to provide a crystalline form of oxalate Form 1 of the compound of formula (I) (hereinafter referred to as oxalate Form 1), using Cu-K ⁇ radiation, and the oxalate Form 1 has an XRPD spectrum substantially the same as that of Figure 55.
  • Another aspect of the present application is to provide a crystalline form of oxalate Form 2 of the compound of formula (I) (hereinafter referred to as oxalate Form 2).
  • oxalate Form 2 a crystalline form of oxalate Form 2 of the compound of formula (I) (hereinafter referred to as oxalate Form 2).
  • the XRPD spectrum of the oxalate Form 2 expressed in 2 ⁇ angle has a characteristic peak at at least one of 15.5° ⁇ 0.2°, 17.2° ⁇ 0.2° and 23.8° ⁇ 0.2°; preferably, there are two or three characteristic peaks.
  • the XRPD spectrum of the oxalate Form 2 also has characteristic peaks at at least one of 13.9° ⁇ 0.2°, 16.9° ⁇ 0.2°, 19.8° ⁇ 0.2° and 16.0° ⁇ 0.2°2 ⁇ .
  • the oxalate Form 2 has an XRPD pattern substantially the same as that of Figure 56.
  • Another aspect of the present application is to provide a method for preparing oxalate Form 2, which comprises: dissolving the compound of formula (I) and oxalic acid in ketone solvents respectively, mixing the two solutions, stirring at room temperature, precipitating, adding an ether solvent, stirring, centrifuging, and drying to obtain oxalate Form 2.
  • the ketone solvent is acetone
  • the ether solvent is isopropyl ether.
  • the mass volume ratio (mg/ml) of the compound of formula (I) to acetone is >100:1.
  • the molar ratio of the compound of formula (I) to oxalic acid is 1:1-1.5.
  • the low temperature stirring temperature is 4°C.
  • Another aspect of the present application is to provide a pharmaceutical composition, which comprises a therapeutically effective amount of a salt of a compound of formula (I) or a crystalline form thereof, and at least one pharmaceutically acceptable carrier.
  • Another aspect of the present application is to provide a preparation prepared from the above-mentioned pharmaceutical composition, wherein the preparation form includes but is not limited to oral solid preparations, injections, and external preparations.
  • the preparation is in the form of tablets, capsules, powder injections, powders, syrups, solutions, suspensions, aerosols or suppositories.
  • the various dosage forms of the present application can be prepared according to conventional preparation methods in the pharmaceutical field.
  • the pharmaceutically acceptable carrier is an excipient commonly used in preparations in the art, including but not limited to any one of adhesives, surfactants, diluents, antiadhesives, hydrophilic or hydrophobic polymers, tranquilizers or stabilizers, disintegrants, antioxidants, defoaming agents, fillers, glidants/lubricants, adsorbents, preservatives, plasticizers, sweeteners, and mixtures of two or more thereof.
  • adhesives including but not limited to any one of adhesives, surfactants, diluents, antiadhesives, hydrophilic or hydrophobic polymers, tranquilizers or stabilizers, disintegrants, antioxidants, defoaming agents, fillers, glidants/lubricants, adsorbents, preservatives, plasticizers, sweeteners, and mixtures of two or more thereof.
  • the filler or diluent is selected from any one or a combination of lactose, D-mannitol, microcrystalline cellulose, starch, pregelatinized starch, calcium sulfate, calcium hydrogen phosphate, and calcium carbonate;
  • the disintegrant is selected from any one or a combination of sodium carboxymethyl starch, sodium hydroxymethyl cellulose, cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, and cross-linked polyvinyl pyrrolidone;
  • the lubricant/glidant is selected from any one or a combination of magnesium stearate, talc, and micro-powdered silica gel.
  • the pharmaceutical composition may also include one or more pH adjusters or buffers, for example: acids, such as any one or a combination of acetic acid, boric acid, citric acid, fumaric acid, maleic acid, tartaric acid, malic acid, lactic acid, phosphoric acid, and hydrochloric acid; or bases, such as any one or a combination of sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane; or buffers, such as citrate/glucose, sodium bicarbonate, ammonium chloride, and the like; such buffers used as bases may have counter ions other than sodium, such as potassium, magnesium, calcium, ammonium, and other counter ions; and other amounts required to maintain the pH of the components within an acceptable range, comprising solutions or solids of such acids, bases, and buffers.
  • acids such as any one or a combination of acetic acid, boric acid, citric acid, fumaric acid, maleic acid
  • Another aspect of the present application is to provide a use of a salt of the compound of formula (I) or a crystal form thereof, or a pharmaceutical composition thereof, in the preparation of a drug for preventing and/or treating diseases caused or regulated by activation of the bifunctional phosphorylation site of STAT3.
  • the application is to inhibit the proliferation, growth, migration, infiltration, cloning and metastasis of cancer cells, promote the apoptosis of cancer cells, and/or prolong the survival of tumor patients.
  • the diseases include but are not limited to examples of malignant tumors, including astroglioma, malignant medulloblastoma, germ cell tumor, craniopharyngioma, ependymoma and other pediatric brain tumors; glioma, meningioma, pituitary adenoma, neurothecoma and other adult brain tumors; maxillary sinus cancer, pharyngeal cancer (nasopharyngeal cancer, middle pharyngeal cancer, laryngeal cancer), oral cancer, lip cancer, tongue cancer, parotid gland cancer and other head and neck cancers; small cell lung cancer, non-small cell lung cancer, thymoma, mesothelioma and other chest cancers and tumors; esophageal cancer, liver cancer, primary liver cancer, gallbladder cancer, bile duct cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, an
  • the disease is selected from pancreatic cancer Capan-2, PANC-1, MIAPACA-2, BXPC3, SW1990, CFPAC-1, Pan02, ASPC-1, gastric cancer MKN45, BGC823, MGC803, liver cancer Huh7, PLC/PRF-5, HepG2, SK-hep1, SMMC7721, HCCC9810, lung cancer HCC827, A549, H460, H23, H1299, H522, H1975, PC-9, prostate cancer DU145, LNCAP, colon cancer HCT8, HT29, HCT-116.
  • Another aspect of the present application is to provide a use of a salt of the compound of formula (I) or a crystal form thereof, or a pharmaceutical composition thereof, in the preparation of an inhibitor that inhibits STAT3 phosphorylation, inhibits STAT3 transcriptional activity and mitochondrial oxidative phosphorylation.
  • Another aspect of the present application is to provide a method for preventing and/or treating diseases related to activation of the bifunctional phosphorylation site of STAT3, which comprises administering an effective amount of a salt of the compound of formula (I) or a crystalline form thereof, or a pharmaceutical composition thereof, to an individual in need.
  • the diseases include but are not limited to examples of malignant tumors, including astroglioma, malignant medulloblastoma, germ cell tumor, craniopharyngioma, ependymoma and other pediatric brain tumors; glioma, meningioma, pituitary adenoma, neurothecoma and other adult brain tumors; maxillary sinus cancer, pharyngeal cancer (nasopharyngeal cancer, middle pharyngeal cancer, laryngeal cancer), oral cancer, lip cancer, tongue cancer, parotid gland cancer and other head and neck cancers; small cell lung cancer, non-small cell lung cancer, thymoma, mesothelioma and other chest cancers and tumors; esophageal cancer, liver cancer, primary liver cancer, gallbladder cancer, bile duct cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, an
  • the disease is selected from pancreatic cancer Capan-2, PANC-1, MIAPACA-2, BXPC3, SW1990, CFPAC-1, Pan02, ASPC-1, gastric cancer MKN45, BGC823, MGC803, liver cancer Huh7, PLC/PRF-5, HepG2, SK-hep1, SMMC7721, HCCC9810, lung cancer HCC827, A549, H460, H23, H1299, H522, H1975, PC-9, prostate cancer DU145, LNCAP, colon cancer HCT8, HT29 and HCT-116.
  • Another aspect of the present application is to provide a salt of the compound of formula (I) or its crystal form and its pharmaceutical composition for combined use with other drugs.
  • the other drug is a drug for preventing and/or treating cancer, renal fibrosis, pulmonary fibrosis, rheumatoid arthritis, psoriasis, lupus erythematosus, inflammatory lung disease, inflammatory bowel disease and the like.
  • the experimental operating temperature generally refers to room temperature, and "room temperature” refers to the temperature between 10°C and 30°C.
  • crystal or “crystalline form” refers to the one confirmed by the X-ray powder diffraction pattern shown. It is well known to those skilled in the art that the experimental error depends on the instrument conditions, sample preparation and sample purity, especially the X-ray powder diffraction pattern will usually change with the instrument conditions. It should be noted that the relative intensity of the X-ray powder diffraction pattern may also change with the experimental conditions, so the order of peak intensity cannot be used as the only or decisive factor. In addition, the experimental error of the peak angle is usually 5% or less, and the error of these angles should also be taken into account, and an error of ⁇ 0.2° is usually allowed.
  • any crystal form having the same or similar characteristic peaks in the X-ray powder diffraction spectrum of the crystal form of the present application belongs to the scope of the present application.
  • “Stirring” can adopt conventional methods in the art, such as stirring methods including magnetic stirring and mechanical stirring, and the stirring speed is 50 to 1800 rpm, preferably 300 to 900 rpm.
  • Separatation can be carried out by conventional methods in the art, such as centrifugation or filtration.
  • the filtration is carried out under reduced pressure, generally at a pressure less than atmospheric pressure, preferably at a pressure less than 0.09 MPa.
  • Drying can be accomplished by conventional techniques in the art, such as room temperature drying, forced air drying or reduced pressure drying; it can be reduced pressure or normal pressure, preferably the pressure is less than 0.09 MPa.
  • the drying apparatus and method are not limited, and can be a fume hood, forced air oven, spray dryer, fluidized bed drying or vacuum oven; it can be carried out under reduced pressure or not, preferably the pressure is less than 0.09 MPa.
  • ratios involved in this application are mass-to-volume ratios between liquid and solid, and volume ratios between liquids.
  • substantially in crystalline form means that the crystallinity determined by X-ray powder diffraction data is greater than about 20%, more preferably greater than about 60%, even greater than about 80%, and more preferably greater than about 90%.
  • Figure 1 is the XRPD spectrum of hemifumarate Form 1 in Example 2;
  • Figure 2 is a TGA spectrum of hemifumarate Form 1 in Example 2;
  • Figure 3 is a DSC spectrum of hemifumarate Form 1 in Example 2;
  • Figure 4 is a DVS spectrum of hemifumarate Form 1 in Example 2;
  • FIG5 is a solid-state 1 H-NMR spectrum of the hemifumarate salt Form 1 in Example 2;
  • Figure 6 is the XRPD spectrum of hemifumarate Form 2 in Example 2.
  • Figure 7 is a TGA spectrum of hemifumarate Form 2 in Example 2.
  • Figure 8 is a DSC spectrum of hemifumarate Form 2 in Example 2.
  • Figure 9 is a DVS spectrum of hemifumarate Form 2 in Example 2.
  • FIG10 is a solid-state 1 H-NMR spectrum of the hemifumarate salt Form 2 in Example 2;
  • Figure 11 is the XRPD spectrum of hemifumarate Form 3 in Example 35;
  • Figure 12 is a TGA spectrum of hemifumarate Form 3 in Example 35;
  • Figure 13 is a DSC spectrum of hemifumarate Form 3 in Example 35;
  • FIG14 is a solid-state 1 H-NMR spectrum of the hemifumarate salt Form 3 in Example 35;
  • Figure 15 is the XRPD spectrum of hemifumarate Form 4 in Example 36;
  • Figure 16 is a TGA spectrum of hemifumarate Form 4 in Example 36;
  • Figure 17 is a DSC spectrum of hemifumarate Form 4 in Example 36;
  • Figure 18 is a DVS spectrum of hemifumarate Form 4 in Example 36;
  • FIG19 is a solid-state 1 H-NMR spectrum of the hemifumarate salt Form 4 in Example 36;
  • FIG20 is a single crystal simulation spectrum of hemifumarate Form 1 in Example 44;
  • Figure 21 is the XRPD spectrum of L-tartrate Form 1 in Example 46;
  • Figure 22 is a TGA spectrum of L-tartrate Form 1 in Example 46;
  • Figure 23 is a DSC spectrum of L-tartrate Form 1 in Example 46;
  • Figure 24 is a DVS spectrum of L-tartrate Form 1 in Example 46;
  • FIG25 is a solid-state 1 H-NMR spectrum of L-tartrate salt Form 1 in Example 46;
  • Figure 26 is the XRPD spectrum of L-tartrate Form 2 in Example 57;
  • Figure 27 is a TGA spectrum of L-tartrate Form 2 in Example 57;
  • Figure 28 is a DSC spectrum of L-tartrate Form 2 in Example 57;
  • FIG29 is a solid-state 1 H-NMR spectrum of L-tartrate salt Form 2 in Example 57;
  • Figure 30 is the XRPD spectrum of L-tartrate Form 3 in Example 58;
  • Figure 31 is a TGA spectrum of L-tartrate Form 3 in Example 58;
  • Figure 32 is a DSC spectrum of L-tartrate Form 3 in Example 58;
  • Figure 33 is a solid-state 1H-NMR spectrum of L-tartrate salt Form 3 in Example 58;
  • Figure 34 is an XRPD spectrum of L-tartrate Form 4 in Example 59;
  • Figure 35 is a TGA spectrum of L-tartrate Form 4 in Example 59;
  • Figure 36 is a DSC spectrum of L-tartrate Form 4 in Example 59;
  • FIG37 is a solid-state 1 H-NMR spectrum of L-tartrate salt Form 4 in Example 59;
  • Figure 38 is the XRPD spectrum of L-tartrate salt Form 5 in Example 74;
  • Figure 39 is an XRPD spectrum of L-tartrate Form 6 in Example 75;
  • FIG40 is an XRPD pattern of amorphous L-tartrate salt in Example 76;
  • FIG41 is a TGA spectrum of amorphous L-tartrate salt in Example 76;
  • FIG42 is a DSC spectrum of amorphous L-tartrate salt in Example 76;
  • FIG43 is a DVS spectrum of amorphous L-tartrate in Example 76;
  • FIG44 is a solid-state 1 H-NMR spectrum of the amorphous L-tartrate salt in Example 76;
  • Figure 45 is the XRPD spectrum of hydrochloride Form 1 in Example 77;
  • Figure 46 is a TGA spectrum of hydrochloride Form 1 in Example 77;
  • Figure 47 is a DSC spectrum of hydrochloride Form 1 in Example 77;
  • Figure 48 is a DVS spectrum of hydrochloride Form 1 in Example 77;
  • Figure 49 is the XRPD spectrum of mesylate Form 1 in Example 78;
  • Figure 50 is the XRPD spectrum of mesylate Form 2 in Example 79;
  • Figure 51 is a TGA spectrum of mesylate Form 2 in Example 79;
  • Figure 52 is a DSC spectrum of mesylate Form 2 in Example 79;
  • Figure 53 is a DVS spectrum of mesylate Form 2 in Example 79;
  • Figure 54 is the XRPD spectrum of mesylate Form 3 in Example 80;
  • Figure 55 is the XRPD spectrum of oxalate Form 1 in Example 81;
  • Figure 56 is the XRPD spectrum of oxalate Form 2 in Example 82;
  • Figure 57 is a TGA spectrum of oxalate Form 2 in Example 82;
  • Figure 58 is a DSC spectrum of oxalate Form 2 in Example 82;
  • Figure 59 is a DVS spectrum of oxalate Form 2 in Example 82;
  • FIG60 is an XRPD overlay of the hemifumarate Form 1 in Experimental Example 1 before and after being placed under high temperature (40°C and 60°C);
  • FIG61 is an XRPD overlay of the hemifumarate Form 1 in Experimental Example 1 before and after being placed under high humidity (25°C/90%RH) and accelerated (40°C/75%RH) conditions;
  • FIG62 is an XRPD overlay of the hemifumarate Form 2 in Experimental Example 1 before and after being placed under high temperature (open, 60°C, ⁇ 30% RH) conditions;
  • FIG63 is an XRPD overlay of L-tartrate Form 1 in Experimental Example 1 before and after long-term (25°C/60%RH) and accelerated (40°C/75%RH) conditions;
  • Figure 64 is an XRPD overlay of L-tartrate Form 1 in Experimental Example 1 before and after being placed under high temperature (60°C, ⁇ 30% RH) conditions;
  • Figure 65 is an XRPD overlay of L-tartrate Form 4 in Experimental Example 1 before and after long-term (25°C/60%RH) and accelerated (40°C/75%RH) conditions;
  • Figure 66 is an XRPD overlay of L-tartrate Form 4 in Experimental Example 1 before and after being placed at 40°C;
  • Figure 67 is an XRPD overlay of L-tartrate Form 4 in Experimental Example 1 before and after being placed at 50°C;
  • Figure 68 is an XRPD overlay of L-tartrate Form 4 in Experimental Example 1 before and after being placed at 55°C;
  • Figure 69 is a PSD graph of hemifumarate Form 1 in Experimental Example 2;
  • Figure 70 is a PSD graph of hemifumarate Form 2 in Experimental Example 2;
  • Figure 71 is the PSD graph of oxalate Form 2 in Experimental Example 2.
  • X-ray powder diffraction (XRPD) data were collected from a Bruker D8 Advance diffractometer; the parameters were as follows: Cu target; wavelength Current and voltage: 40KV, 40mA; Angle range: 3 ⁇ 40°2 ⁇ .
  • thermogravimetric analysis (TGA) data were collected from TA Instruments Q500TGA; the parameters were as follows: mode: high resolution mode; heating rate: 10°C/min; protective gas: N2; sample pan: platinum crucible.
  • DSC differential thermal analysis
  • vacuum drying adopts vacuum drying oven, model is D2F-6020.
  • the analytical balance model is BT125D.
  • HPLC high performance liquid chromatography
  • HPLC high performance liquid chromatography
  • the starting material compound of formula (I) can be prepared by the method mentioned in CN112300145A.
  • the results are as follows:
  • the results are as follows:
  • the results are as follows:
  • the hemifumarate of the compound of formula (I) (the compound of formula (II)) can be prepared in multiple solvent systems, and most of them are in a crystalline form with good crystallinity. Under the same conditions, most other acids cannot form salts, or the salts formed are in an amorphous state or have poor crystallinity.
  • Example 2 Preparation of the hemifumarate salt of the compound of formula (I) in crystalline form
  • the hemifumarate of the compound of formula (I) in crystalline form is exposed to an environment with a relative humidity of 75% RH and a temperature of 30°C for 2 days to obtain hemifumarate Form 1.
  • the hemifumarate of the compound of formula (I) in crystalline form is exposed to an environment with a relative humidity of ⁇ 10% RH and a temperature of 30°C for 2 days to obtain hemifumarate Form 2.
  • Example 3-5 Preparation of the hemifumarate salt of the compound of formula (I) in crystalline form
  • Example 6 Preparation of the hemifumarate salt of the compound of formula (I) in crystalline form
  • Example 7 Preparation of the hemifumarate salt of the compound of formula (I) in crystalline form
  • Example 11-34 Preparation of the hemifumarate salt of the compound of formula (I) in crystalline form
  • hemifumarate Form 2 Take about 100 mg of hemifumarate Form 2, put it in a centrifuge tube, place it in a bottle filled with ethanol at room temperature for 6 days, and after testing, hemifumarate Form 3 was obtained.
  • L-tartrate Form 2 Take about 20 mg of L-tartrate Form 1 into a 3 ml vial and place it under RT-97% relative humidity conditions for 7 days to obtain L-tartrate Form 2.
  • L-tartrate salt Form 1 About 20 mg of L-tartrate salt Form 1 was suspended in 0.5 mL of a mixed solvent of dimethyltetrahydrofuran and an alkane solvent in a volume ratio of 1:4. The suspension was stirred at 4-8°C for about 3 days, and the solid was separated to obtain L-tartrate salt Form 3.
  • L-tartrate Form 1 Take about 200 mg of L-tartrate Form 1 and dissolve it in 1 mL of a mixed solvent of acetone and methyl tert-butyl ether in a volume ratio of 1:1, filter to obtain a clear solution, and then put the clear solution into a 20 mL vial sealed with a sealing film with a pinhole on the sealing film. Leave it at room temperature to allow the pinhole to evaporate slowly. After 1 day, a solid precipitates and is separated to obtain L-tartrate Form 4.
  • L-tartrate Form 1 Take about 1.0 g of L-tartrate Form 1, suspend it in 25 mL of a mixed solvent of acetone/n-heptane with a volume ratio of 1:6, and introduce a small amount of L-tartrate Form 4 as a seed crystal. The suspension agglomerates and stirs at room temperature overnight (about 16 hours). Separate the solid to obtain L-tartrate Form 4.
  • L-tartrate Form 4 About 1.5 g of the compound of formula (I) and about 320.58 mg of L-tartaric acid are suspended in 50 mL of a 1:6 acetone/n-heptane mixed solvent, and a small amount of L-tartrate Form 4 is introduced as a seed crystal. The mixture is stirred at room temperature for about 2 days, and the solid is separated to obtain L-tartrate Form 4.
  • L-tartrate Form 1 Take about 20 mg of L-tartrate Form 1 and suspend it in 0.5 mL of methyl tert-butyl ether. After stirring the suspension at 4-8°C for about 3 days, separate the solid to obtain L-tartrate Form 4.
  • L-tartrate salt Form 1 About 20 mg was suspended in 0.5 mL of a mixed solvent of butanone and n-heptane in a volume ratio of 1:6. After the suspension was stirred at room temperature for about 3 days, the solid was separated to obtain L-tartrate salt Form 4.
  • L-tartrate Form 1 Take about 20 mg of L-tartrate Form 1 into a 3 mL vial, place it in a 20 mL vial containing 3 mL of acetone or ethyl acetate, seal the 20 mL vial with a lid, and keep it at room temperature for 7 days to allow the solvent vapor to interact with the sample and separate the solid to obtain L-tartrate Form 4.
  • L-tartrate Form 1 Take about 20 mg of L-tartrate Form 1 and dissolve it in 0.1-3.0 mL of ethyl acetate. Filter the filtrate and add about 1-2 mg of ethyl cellulose into a vial. Seal the vial containing the filtrate with a sealing film and pierce a small hole. Slowly evaporate at room temperature to separate the solid and obtain L-tartrate Form 4.
  • L-tartrate salt Form 1 About 20 mg of L-tartrate salt Form 1 was suspended in 0.5 mL of a 1:1 volume ratio isopropanol/methyl tert-butyl ether mixed solvent. After the suspension was stirred at room temperature for about 3 days, the solid was separated to obtain L-tartrate salt Form 5.
  • the high performance liquid chromatography (HPLC) detection result shows that the molar ratio of the compound of formula (I) to hydrochloric acid is 1:1.
  • L-tartrate Form 1 is placed under long-term (25°C/60%RH) and accelerated (40°C/75%RH) conditions for 1 week, and the crystal form and purity remain basically unchanged, showing good crystal stability and chemical stability.
  • L-tartrate Form 1 is placed under high temperature (60°C, ⁇ 30%RH) conditions for 10 days, and its crystal form remains basically unchanged (the results are shown in Figure 64), and the increase of related substances is ⁇ 0.05%, showing good crystal stability and chemical stability.
  • L-tartrate Form 4 remains basically unchanged in crystal form and purity after being placed under long-term (25°C/60%RH) and accelerated (40°C/75%RH) conditions for 1 week, and has good crystal stability and chemical stability.
  • an appropriate amount of L-tartrate Form 4 was placed at high temperatures of 40°C, 50°C and 55°C for 30 days in two conditions: with packaging materials and exposed, and its crystal form remained basically unchanged (the results are shown in Figures 66-68), and it has good crystal stability.
  • the repose angle of the crystal form was tested using the HYL-1001 multifunctional powder physical property analyzer of Haoyu Technology Co., Ltd. The results showed that the repose angle of oxalate Form 2 was 58.10°, and the repose angles of L-tartrate Form 1 and L-tartrate Form 4 were 40.20° and 32.73°, respectively. According to the evaluation criteria of powder indicators in Table 6, the fluidity of L-tartrate Form 4 and L-tartrate Form 1 were better than that of oxalate Form 2.
  • Tablets containing each salt form and its crystal form were prepared according to the tablet formulation in Table 8. Specifically, the components were mixed, vortexed and mixed, and all the mixtures were pressed (pressed into tablets at a pressure of 2 MPa for 2 min) using an infrared tablet press to prepare tablets.
  • the prepared hemi-fumarate Form 1 tablets, hemi-fumarate Form 2 tablets and oxalate Form 2 tablets were taken respectively, and the diameter and hardness were tested with a hardness tester, the thickness was tested with a vernier caliper, and the tensile strength was calculated.
  • the results showed that the average tensile strength of hemi-fumarate Form 1 tablets was 1.20MPa, the average tensile strength of hemi-fumarate Form 2 was 1.34MPa, and the average tensile strength of oxalate Form 2 was 1.14MPa. Under a certain pressure, the greater the tensile strength, the better the compressibility. Therefore, compared with the tablets prepared from oxalate Form 2, the tablets prepared from hemi-fumarate Form 1 and the tablets prepared from hemi-fumarate Form 2 have better compressibility.
  • Dissolution medium pH 1.2 solution (containing 0.5% Tween 80)
  • Dissolution method Paddle method, 75rpm
  • Gender and age Male, 6 to 8 weeks.
  • the blood concentration and curve area of hemifumarate Form 1, hemifumarate Form 2 and L-tartrate Form 4 have obvious advantages over the free state of the compound of formula (I), and the oral absolute bioavailability of hemifumarate Form 1, hemifumarate Form 2 and L-tartrate Form 4 has obvious advantages over the free state of the compound of formula (I).
  • Gender and age Male, 6 to 8 weeks.
  • the blood concentration and curve area of L-tartrate Form 1 have obvious advantages over the free state of the compound of formula (I), and the oral absolute bioavailability of L-tartrate Form 1 has obvious advantages over the free state of the compound of formula (I).
  • mice Animal species and strain: Balb/c nude mice
  • Gender and age Female, 7-9 weeks old.
  • Preparation and storage of dosing solution Weigh a certain amount of hemi-fumarate Form 1 (based on the free base content) into a suitable container, add the calculated amount of solvent, stir, vortex or ultrasonically mix to obtain the dosing solution of desired concentration, record the physical properties, and store at 4°C for use immediately after preparation.
  • MDA-MB-231 cells were cultured in L-15 medium containing 10% fetal bovine serum (FBS) (GIBCO, USA) in a closed culture flask without carbon dioxide and placed in an incubator at 37°C.
  • FBS fetal bovine serum
  • MDA-MB-231 cells in the logarithmic growth phase were collected, resuspended in serum-free L-15 basal medium, added to Matrigel at a ratio of 1:1, and the concentration of the cell suspension was adjusted to 6.67 ⁇ 107/ml. Under sterile conditions, 0.15 mL of the cell suspension was inoculated subcutaneously on the right back of the mouse, with an inoculation concentration of 1 ⁇ 107/0.15 mL/mouse.
  • the animals were randomly divided into groups according to the tumor volume so that the difference in tumor volume between groups was less than 10% of the mean.
  • the day of grouping was designated as Day 0, and drug administration was started according to the animal weight or the fixed dosage volume for each animal.
  • the dosing schedule is shown in Table 14.
  • the experimental period was 4 weeks. During the experiment, the animal body weight and tumor volume were measured twice a week, and the clinical symptoms of the animals were observed and recorded once a day.
  • the calculation formula for tumor volume (TV) is: 1/2 ⁇ a ⁇ b2, where a and b are the measured length and width of the tumor, respectively.
  • the relative tumor volume (RTV) calculation formula is: Vt/V0, where V0 is the tumor volume at the time of grouping and Vt is the tumor volume at each measurement.
  • the relative tumor proliferation rate T/C was calculated as follows: TRTV/CRTV ⁇ 100%, where TRTV was the RTV of the treatment group and CRTV was the RTV of the negative control group.
  • TGI tumor volume inhibition rate
  • the tumor inhibition rate (%) was calculated as follows: (TWC-TWT)/TWC ⁇ 100%, where TWC was the average tumor weight of the negative control group and TWT was the average tumor weight of the treatment group.
  • BWC (%) (BWt-BW0)/BW0 ⁇ 100%, where BWt is the animal weight at each measurement and BW0 is the animal weight when grouped.
  • the tumor growth curve was drawn with time point as X-axis and tumor volume (mm 3 ) as Y-axis; the animal weight change curve was drawn with time point as X-axis and animal weight (g) as Y-axis.
  • the two-tailed t-test was used for comparison between groups, with P ⁇ 0.05 as significant difference and P ⁇ 0.01 as extremely significant difference.
  • the average tumor volume of the vehicle control group was 564.69 ⁇ 26.44mm 3
  • the average tumor volume of the hemi-fumarate Form 1 (10mg/kg) group was 138.60 ⁇ 14.36mm 3
  • the tumor volume inhibition rate TGI (%) was 75.46%
  • the relative tumor proliferation rate T/C after administration was 23.91%, which was less than 40%, and it could significantly inhibit the growth of human breast cancer MDA-MB-231 cell Balb/c nude mouse xenograft tumors.
  • the average tumor volume of the L-tartrate Form 4 (10mg/kg) group was 116.17 ⁇ 7.25, the tumor volume inhibition rate TGI (%) was 79.43%, and the relative tumor proliferation rate T/C after administration was 20.37%, which was less than 40%, and it could significantly inhibit the growth of human breast cancer MDA-MB-231 cell Balb/c nude mouse xenograft tumors.
  • Gender and age Female, 6-7 weeks old.
  • Preparation and storage of dosing solution Weigh a certain amount of hemi-fumarate Form 1 (based on the free base content) into a suitable container, add the calculated amount of solvent, stir, vortex or ultrasonically mix to obtain the dosing solution of desired concentration, record the physical properties, and prepare and use at room temperature.
  • MOLM16 cells were cultured in RPMI-1640 medium containing 20% FBS and maintained in a 37°C saturated humidity incubator with 5% CO 2 .
  • MOLM16 cells in logarithmic growth phase were collected, resuspended in RPMI-1640 basal medium, and matrigel was added at a ratio of 1:1 to adjust the cell concentration to 1 ⁇ 10 7 /mL.
  • 0.1mL of cell suspension was inoculated subcutaneously on the right back of nude mice at an inoculation concentration of 1 ⁇ 10 6 /0.1mL/mouse.
  • the animals were randomly divided into groups according to the tumor volume so that the difference in tumor volume between groups was less than 10% of the mean.
  • the day of grouping was designated as Day 0, and drug administration was started according to the animal weight or the fixed administration volume for each animal.
  • the administration schedule is shown in Table 16.
  • the experimental period was 2 weeks. During the experiment, the animal body weight and tumor volume were measured twice a week, and the clinical symptoms of the animals were observed and recorded once a day.
  • the average tumor volume of the vehicle control group was 1788.88 ⁇ 132.86mm 3
  • the average tumor volume of the hemi-fumarate Form 1 (1mg/kg, 3mg/kg and 10mg/kg) groups was 172.85 ⁇ 14.02mm 3 , 120.33 ⁇ 3.15mm 3 and 64.08 ⁇ 7.11mm 3 , respectively
  • mice Animal species and strain: CB-17 SCID mice.
  • Gender and age Female, 6-8 weeks old.
  • Body weight 18-22 g, deviation is ⁇ 20% of the mean body weight.
  • Preparation and storage of dosing solution Weigh a certain amount of hemi-fumarate Form 1 (based on the free base content) into a suitable container, add the calculated amount of solvent, stir, vortex or ultrasonically mix to obtain the dosing solution of desired concentration, record the physical properties, and store at 4°C for use immediately after preparation.
  • SU-DHL-1 cells were cultured in RPMI-1640 medium containing 10% FBS and maintained in a 37°C humidified incubator with 5% CO 2 .
  • SU-DHL-1 cells in the logarithmic growth phase were collected, resuspended in RPMI-1640 basal medium, and matrigel was added at a ratio of 1:1 to adjust the cell concentration to 3 ⁇ 10 7 /mL.
  • matrigel was added at a ratio of 1:1 to adjust the cell concentration to 3 ⁇ 10 7 /mL.
  • 0.1 mL of the cell suspension was inoculated subcutaneously on the right back of nude mice at an inoculation concentration of 3 ⁇ 10 6 /0.1 mL/mouse.
  • the animals were randomly divided into groups according to the tumor volume so that the difference in tumor volume between groups was less than 10% of the mean.
  • the day of grouping was designated as Day 0, and drug administration was started according to the animal weight or the fixed administration volume for each animal.
  • the administration schedule is shown in Table 18.
  • the experimental period was 3 weeks. During the experiment, the animal body weight and tumor volume were measured twice a week, and the clinical symptoms of the animals were observed and recorded once a day.
  • the average tumor volume of the vehicle control group was 2029.90 ⁇ 204.72 mm 3
  • the average tumor volume of the hemi-fumarate Form 1 (10 mg/kg) group was 316.83 ⁇ 97.40 mm 3
  • Hemi-fumarate Form 1 could significantly inhibit the growth of human metastatic large cell lymphoma SU-DHL-1 cell CB-17 SCID mouse xenograft tumors.
  • mice Animal species and strains: mice, NOD-SCID
  • Gender and age Male, 6-8 weeks.
  • Preparation of test compound Preparation of solvent: Take an appropriate amount of Tween80 and add 99ml of 0.5% MC, vortex mix, and set aside;
  • Preparation of dosing solution Weigh a certain amount of the free form of the compound of formula (I) and L-tartrate Form 1 (based on the content of free base) into a suitable container, add a calculated amount of solvent, stir and mix to obtain a dosing solution of the desired concentration, which is prepared and used immediately.
  • Molm16 cells were cultured in 1640 medium containing 20% inactivated fetal bovine serum (Gbico-10099141) and 1% penicillin/streptomycin, and cultured continuously in a cell culture flask in a cell culture incubator at 37°C containing 5% CO2; when the cells grew to the logarithmic growth phase, the cells were collected and washed once with PBS, the cells were counted, centrifuged, and finally resuspended with PBS, 50% Matrigel (Cultrex PathClear BM, 3432-010-01P) was added, and the cell concentration was adjusted for subsequent inoculation; 3 ⁇ 10 6 cells were inoculated in the right axilla of each mouse, and the inoculation volume was 100ul.
  • Gbico-10099141 inactivated fetal bovine serum
  • penicillin/streptomycin penicillin/streptomycin
  • the tumor volume calculation formula: 0.5a ⁇ b 2
  • a and b represent the length and width of the tumor, respectively, and the tumor volume was used to calculate the tumor volume, and the grouping was based on the tumor volume.
  • Tumors with a volume of 80 mm 3 to 120 mm 3 were randomly grouped, and the mean tumor volume of each group was 93 mm 3 , and then the drug was administered.
  • the drug administration scheme is shown in Table 20.
  • the health status and mortality of animals were monitored daily. Routine inspections included observing the effects of the test substances and drugs on the daily behavior of animals, such as behavioral activities, food and water intake, body weight changes (body weight was measured twice a week), physical signs or other abnormal conditions.
  • Tumor diameters were measured twice a week, and tumor volumes were calculated.
  • TGI% (1-T/C) ⁇ 100%
  • Two-way ANOVA was used to compare body weight and tumor volume.
  • One-way ANOVA was used to compare tumor weight at the end point of the experiment. All data will be analyzed using GraphPad Prism 8, and p ⁇ 0.05 indicated statistical significance.
  • Experimental Example 13 Study on the inhibitory effect of L-tartrate Form 4 on the growth of human pancreatic cancer BxPC-3 cell Balb/c nude mouse xenograft tumors.
  • Experimental purpose To evaluate the anti-tumor activity of L-tartrate Form 4 using a human pancreatic cancer BxPC-3 cell Balb/c nude mouse xenograft tumor model.
  • mice Animal species and strain: Balb/c nude mice
  • Gender and age Female, 9-11 weeks old.
  • Body weight 21-24 g, with a deviation of ⁇ 20% from the mean body weight.
  • Solution preparation and storage Preparation of solvent (0.5% MC solution): Pipette an appropriate amount of MC into a suitable container, add a calculated amount of ddH2O, stir, vortex or sonicate to fully dissolve, and seal for later use.
  • Preparation and storage of dosing solution Weigh a certain amount of L-tartrate Form 4 (based on the content of free base) into a suitable container, add the calculated amount of solvent, stir, vortex or ultrasonically mix to obtain the desired concentration of dosing solution, and record the physical properties. 4°C, prepare and use immediately.
  • BxPC-3 cells were cultured in RPMI-1640 medium containing 10% FBS and maintained in a 37°C saturated humidity incubator with 5% CO 2 .
  • BxPC-3 cells in logarithmic growth phase were collected, resuspended in RPMI-1640 basal medium, and matrigel was added at a ratio of 1:1 to adjust the cell concentration to 5 ⁇ 10 7 /mL.
  • matrigel was added at a ratio of 1:1 to adjust the cell concentration to 5 ⁇ 10 7 /mL.
  • 0.1 mL of cell suspension was inoculated subcutaneously on the right back of nude mice at an inoculation concentration of 5 ⁇ 10 6 /0.1 mL/mouse.
  • the animals were randomly divided into groups according to the tumor volume so that the difference in tumor volume between groups was less than 10% of the mean.
  • the day of grouping was designated as Day 0, and drug administration was started according to the animal weight or the fixed administration volume for each animal.
  • the administration schedule is shown in Table 22.
  • the experimental period was 4 weeks. During the experiment, the animal body weight and tumor volume were measured twice a week, and the clinical symptoms of the animals were observed and recorded once a day.
  • the average tumor volume of the vehicle control group was 1175.00 ⁇ 147.32mm 3
  • the average tumor volume of the 3mg/kg group and the 10mg/kg group of L-tartrate Form 4 was 379.47 ⁇ 65.56mm 3 and 307.27 ⁇ 49.38mm 3 , respectively.
  • the tumor volume inhibition rate TGI (%) was 67.70% and 73.85%, respectively.
  • the relative tumor proliferation rate T/C after administration was 32.44% and 26.78%, respectively, which was less than 40%.
  • L-tartrate Form 4 can significantly inhibit the growth of human pancreatic cancer BxPC-3 cell Balb/c nude mouse xenograft tumors.
  • mice Animal species and strain: Balb/c nude mice
  • Gender and age female, 8-9 weeks old.
  • Body weight 18-22 g, deviation is ⁇ 20% of the mean body weight.
  • Solution preparation and storage Preparation of solvent (0.5% MC solution): Pipette an appropriate amount of MC into a suitable container, add a calculated amount of ddH 2 O, stir, vortex or ultrasonicate to fully dissolve, and seal for later use.
  • Preparation and storage of dosing solution Weigh a certain amount of L-tartrate Form 4 (based on the content of free base) into a suitable container, add the calculated amount of solvent, stir, vortex or ultrasonically mix to obtain the desired concentration of dosing solution, and record the physical properties. 4°C, prepare and use immediately.
  • NCI-H460 cells were cultured in RPMI-1640 medium containing 10% FBS and maintained in a 37°C saturated humidity incubator with 5% CO2.
  • NCI-H460 cells in the logarithmic growth phase, resuspend them in RPMI-1640 basal medium, add matrix gel at a ratio of 1:1, and adjust the cell concentration to 2 ⁇ 107/mL. Under sterile conditions, inoculate 0.1mL of the cell suspension subcutaneously on the right back of nude mice at an inoculation concentration of 2 ⁇ 106/0.1mL/mouse.
  • the animals were randomly divided into groups according to the tumor volume so that the difference in tumor volume between groups was less than 10% of the mean.
  • the day of grouping was designated as Day 0, and drug administration was started according to the animal weight or the fixed administration volume for each animal.
  • the administration schedule is shown in Table 24.
  • the experimental period was 17 days. During the experiment, the animal body weight and tumor volume were measured twice a week, and the clinical symptoms of the animals were observed and recorded once a day.
  • the average tumor volume of the vehicle control group was 2109.69 ⁇ 113.70 mm 3
  • the average tumor volume of the 3 mg/kg group and the 10 mg/kg group of L-tartrate Form 4 were 803.58 ⁇ 42.42 and 710.49 ⁇ 52.36, respectively
  • the tumor volume inhibition rate TGI (%) was 61.91% and 66.32%, respectively
  • the relative tumor growth rate TGI (%) after administration was 38.29% and 33.68%, respectively, which was less than 40%.
  • L-tartrate Form 4 can significantly inhibit the growth of human non-small cell lung cancer NCI-H460 cell Balb/c nude mouse xenograft tumors.

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Abstract

本申请涉及药物化学领域。具体而言，本申请涉及一种三芳香环类衍生物，具体为(6-((5-(3-(4-(三氟甲基)苯基)-1,2,4-噁二唑-5-基)吡嗪-2-基)氧基)-1-甲基-1H-吲哚-2-基)(4-(4-(2,2,2-三氟乙氧基)苄基)哌嗪-1-基)甲酮的盐、其晶型及其药物组合物、制备方法和用途。本申请提供的盐及其晶型稳定性好，溶解度高，生物利用度高，药效好，片剂溶出好，片剂可压性好，流动性好，晶体形貌好，晶体粒度分布好，能稳定储存，制备方法简单可靠，具有较大的开发价值。

Description

三芳香环类衍生物的盐、其晶型、制备方法和用途
相关申请的引用
本申请要求于2023年3月27日向中华人民共和国国家知识产权局提交的申请号为202310307084.2、202310307025.5和202310306887.6的发明专利申请的全部权益，并通过引用的方式将其全部内容并入本申请。
技术领域
本申请涉及药物化学领域。具体而言，本申请涉及一种三芳香环类衍生物，具体为(6-((5-(3-(4-(三氟甲基)苯基)-1,2,4-噁二唑-5-基)吡嗪-2-基)氧基)-1-甲基-1H-吲哚-2-基)(4-(4-(2,2,2-三氟乙氧基)苄基)哌嗪-1-基)甲酮的盐、其晶型及其药物组合物、制备方法和用途。
背景技术
中国专利申请CN112300145A公开了一种三芳香环类化合物，其化学名称为(6-((5-(3-(4-(三氟甲基)苯基)-1,2,4-噁二唑-5-基)吡嗪-2-基)氧基)-1-甲基-1H-吲哚-2-基)(4-(4-(2,2,2-三氟乙氧基)苄基)哌嗪-1-基)甲酮，结构式如式(I)所示：
式(I)化合物作为靶向STAT3双功能磷酸化位点的抑制剂，用于预防和/或治疗与STAT3双功能磷酸化位点活化引起或调控的疾病，具有良好的临床前景。
对于不同的药物活性分子，特别是口服固体药物，不同的固体形式(盐、晶型)往往影响到药物的稳定性、溶解度、纯度、溶出、以及生物利用度等，并且仅仅根据分子结构，通常不能精确合理的预测其是否可以成盐、成盐后是否可以结晶、以及结晶后的性质如何等。例如，一些盐在环境温度下是固体，而其他盐在环境温度下是液体或油状物。此外，一些盐形式在极端条件(比如加热)可以保持稳定，而其他盐在常规条件(比如室温)易于分解。因此，研发用于不同药物活性分子的成盐形式是不可预测的过程。另外，化合物的盐可能以一种或多种晶型存在，但是也无法具体预期其存在与特性。
因此在后续的开发过程中，为了使产品易于生产、便于储存、产品长期稳定、且可药用等，有必要对式(I)化合物的盐及盐的晶型进行深入全面的研究。
发明内容
中国专利申请CN112300145A所涉及的所有内容均以引证的方式添加到本申请中。
本申请提供了式(I)化合物的盐和盐的晶型。本申请提供的盐和盐的晶型至少具有以下一个优点：稳定性好，溶解度高，生物利用度高，药效好，片剂溶出好，片剂可压性好，流动性好，晶体形貌好，晶体粒度分布好，能稳定储存，制备方法简单可靠，具有较大的开发价值。
本申请的一个方面，在于提供一种式(I)化合物的盐，
所述盐为酸式盐，其中，所述式(I)化合物和酸的摩尔比为0.5-2。
本申请优选地技术方案中，所述式(I)化合物的盐选自富马酸盐、L-酒石酸盐、盐酸盐、甲磺酸盐和草酸盐。
本申请优选地技术方案中，所述式(I)化合物的盐选自富马酸盐。
优选地，所述式(I)化合物和富马酸的摩尔比为1:0.5-1；更优选为1:0.5。
本申请优选地技术方案中，所述式(I)化合物的盐为半富马酸盐，结构式如式(II)所示：
优选地，所述式(I)化合物的半富马酸盐基本上以结晶形式或无定型形式存在；更优选为基本上以结晶形式存在。
优选地，所述式(I)化合物半富马酸盐的结晶形式可以是一种或多种；具体地，可以为无水物、水合物或溶剂合物中的至少一种。
本申请的另一个方面，还在于提供一种所述式(I)化合物半富马酸盐的制备方法，其包括将式(I)化合物溶于溶剂1，然后加入富马酸或富马酸溶液反应得到。
本申请优选地技术方案中，所述式(I)化合物半富马酸盐的制备方法包括以下步骤：
步骤A:将适量式(I)化合物在溶剂1中溶解；
步骤B:将适量富马酸在有机溶剂中溶解；
步骤C:将上述两种溶液合并，搅拌析出固体或加入溶剂2后搅拌析出固体；
步骤D:分离出固体得到式(I)化合物半富马酸盐。
优选地，所述溶剂1选自醇类、醚类、腈类、酯类和酮类中的任一种或其组合；优选地，所述醇类选自甲醇、乙醇、正丙醇、仲丁醇、正丁醇、三氟乙醇中的任一种或其组合；所述醚类选自乙醚、甲基叔丁基醚中的任一种或其组合；所述腈类选自乙腈；所述酯类选自乙酸乙酯、乙酸异丙酯和乙酸正丁酯中的任一种或其组合；所述酮类选自丙酮、丁酮中的任一种或其组合。
在一些实施例中，所述溶剂1选自甲醇、乙酸乙酯、乙腈中的任一种或其组合。
优选地，所述有机溶剂选自醇类。
在一些实施例中，所述有机溶剂选自甲醇。
优选地，所述溶剂2选自水、异丙醚、异丙醇、正庚烷和甲基环己烷中的任一种或其组合。
在一些实施例中，所述溶剂2选自水和异丙醚中的任一种或其组合。
优选地，所述式(I)化合物和富马酸的投料摩尔比为1：0.5-1.5；优选地，所述投料摩尔比为1:1。
优选地，所述式(I)化合物和溶剂1的质量体积比(mg/ml)≤100:1；优选≤80:1；更优选≤50:1。
在一些实施例中，(I)化合物和溶剂1的质量体积比(mg/ml)为25:1。
本申请优选地技术方案中，所述式(I)化合物半富马酸盐为结晶形式，使用Cu-Kα辐射，所述半富马酸盐结晶形式以2θ角度表示的X-射线粉末衍射(XRPD)图谱在15.7°±0.2°、16.1°±0.2°和9.4°±0.2°中的至少一处具有特征峰。优选为二处或三处有特征峰。
优选地，所述半富马酸盐结晶形式以2θ角度表示的XRPD图谱在15.7°±0.2°、16.1°±0.2°和9.4°±0.2°处有特征峰。
优选地，所述半富马酸盐结晶形式的XRPD图谱还在14.0°±0.2°、17.1°±0.2°、17.9°±0.2°和19.2°±0.2°2θ中的至少一处具有特征峰。
优选地，所述半富马酸盐结晶形式的XRPD图谱还在18.7°±0.2°、21.8°±0.2°、23.4°±0.2和27.1°±0.2°2θ中的至少一处具有特征峰。
本申请优选地技术方案中，所述半富马酸盐结晶形式为式(I)化合物半富马酸盐Form 1(简称半富马酸盐Form 1)。
优选地，所述半富马酸盐Form 1的XRPD图谱还在7.0°±0.2°、10.2°±0.2°、24.5°±0.2°和20.1°±0.2°2θ中的至少一处具有特征峰。
优选地，所述Form 1的XRPD图谱在7.0°±0.2°、8.2°±0.2°、9.4°±0.2°、9.6°±0.2°、10.2°±0.2°、12.0°±0.2°、14.0°±0.2°、15.7°±0.2°、16.1°±0.2°、17.1°±0.2°、17.9°±0.2°、18.7°±0.2°、19.2°±0.2°、20.1°±0.2°、20.9°±0.2°、21.2°±0.2°、21.8°±0.2°、22.3°±0.2°、22.9°±0.2°、23.4°±0.2°、23.8°±0.2°、24.1°±0.2°、24.5°±0.2°、25.1°±0.2°、25.3°±0.2°、26.5°±0.2°、27.1°±0.2°、27.8°±0.2°、28.5°±0.2°、28.9°±0.2°、29.4°±0.2°、 29.7°±0.2°、31.1°±0.2°、31.2°±0.2°、33.3°±0.2°、34.4°±0.2°和37.6°±0.2°2θ处具有特征峰。
优选地，所述半富马酸盐Form 1具有基本上与图1相同的XRPD图谱。
优选地，所述半富马酸盐Form 1具有基本上与图2相同的TGA图谱。
优选地，所述半富马酸盐Form 1的DSC图谱显示其熔点为156℃±3℃。
优选地，所述半富马酸盐Form 1具有基本上与图3相同的DSC图谱。
优选地，所述半富马酸盐Form 1具有基本上与图4相同的DVS图谱。
优选地，所述半富马酸盐Form 1为无水物。
本申请优选地技术方案中，所述半富马酸盐Form 1的单晶晶体学参数为：单斜晶系，空间群为P21/n；晶胞参数为： α＝γ＝90°，β＝104.031(2)°。
非限制性地，所述半富马酸盐Form 1具有基本如图20所示的XRD单晶模拟图谱。
本申请优选地另一个技术方案中，所述结晶形式为式(I)化合物半富马酸盐Form 2(简称半富马酸盐Form 2)。
优选地，所述半富马酸盐Form 2的XRPD图谱还在16.5°±0.2°、17.3°±0.2°、19.5°±0.2°、24.8°±0.2°和27.6°±0.2°2θ中的至少一处具有特征峰。
优选地，所述半富马酸盐Form 2的XRPD图谱在4.2°±0.2°、5.0°±0.2°、9.3°±0.2°、9.9°±0.2°、11.2°±0.2°、12.6°±0.2°、14.0°±0.2°、15.1°±0.2°、15.6°±0.2°、16.2°±0.2°、16.5°±0.2°、17.0°±0.2°、17.3°±0.2°、18.1°±0.2°、18.4°±0.2°、18.7°±0.2°、19.2°±0.2°、19.5°±0.2°、19.8°±0.2°、20.0°±0.2°、20.4°±0.2°、21.1°±0.2°、21.6°±0.2°、21.9°±0.2°、22.4°±0.2°、22.7°±0.2°、23.5°±0.2°、24.0°±0.2°、24.8°±0.2°、25.1°±0.2°、25.6°±0.2°、26.3°±0.2°、26.9°±0.2°、27.6°±0.2°、28.1°±0.2°、29.0°±0.2°、29.4°±0.2°、30.3°±0.2°、31.3°±0.2°、31.7°±0.2°、33.7°±0.2°、34.6°±0.2°、36.0°±0.2°和38.3±0.2°2θ处具有特征峰。
优选地，所述半富马酸盐Form 2具有基本上与图6相同的XRPD图谱。
优选地，所述半富马酸盐Form 2具有基本上与图7相同的TGA图谱。
优选地，所述半富马酸盐Form 2具有基本上与图8相同的DSC图谱。
优选地，所述半富马酸盐Form 2具有基本上与图9相同的DVS图谱。
优选地，所述半富马酸盐Form 2为无水物。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物半富马酸盐的制备方法，所述制备方法包含以下方法中的任一种：
1)步骤A:将适量式(I)化合物在溶剂3中溶解；
步骤B:将适量富马酸在有机溶剂中溶解；
步骤C:将上述两种溶液合并，搅拌析出或加入溶剂4后搅拌析出，形成混悬液；
步骤D:继续搅拌上述混悬液，并将其在低温环境下搅拌析晶，分离、干燥，得到结晶形式的式(I)化合物半富马酸盐。
优选地，所述溶剂3选自醇类、醚类、腈类、酯类和酮类中的一种或其组合；优选地，所述醇类选自甲醇、乙醇、正丙醇、仲丁醇、正丁醇、三氟乙醇中的任一种或其组合；所述醚类选自乙醚、甲基叔丁基醚中的任一种或其组合；所述腈类选自乙腈；所述酯类选自乙酸乙酯、乙酸异丙酯和乙酸正丁酯中的任一种或其组合；所述酮类选自丙酮、丁酮中的任一种或其组合。
在一些实施例中，所述溶剂3选自甲醇、乙酸乙酯、乙腈中的任一种或其混合。
本申请优选地技术方案中，所述有机溶剂选自醇类。
在一些实施例中，所述有机溶剂选自甲醇。
本申请优选地技术方案中，所述溶剂4选自水、异丙醚、异丙醇、正庚烷和甲基环己烷中的一种或其组合。
在一些实施例中，所述溶剂4选自水和异丙醚中的任一种或其组合。
本申请优选地技术方案中，所述式(I)化合物在溶剂3中加热溶清；所述富马酸在有机溶剂中加热溶清；优选地，所述加热溶清的温度≥40℃；更优选50℃-60℃；
本申请优选地技术方案中，所述步骤C中的搅拌时间≥1h。
本申请优选地技术方案中，所述步骤D中的低温环境为≤30℃；更优选为0℃-20℃。
在一些实施例中，所述步骤D中的低温环境为4℃。
在一些实施例中，所述干燥时间为8-24h；所述干燥的温度为40℃。
本申请优选地技术方案中，所述式(I)化合物和富马酸的投料摩尔比为1：0.5-1.5。
本申请优选地技术方案中，所述式(I)化合物和溶剂3的质量体积比(mg/ml)≤100:1；优选为80:1；更优选为50:1。
在一些实施例中，(I)化合物和溶剂3的质量体积比(mg/ml)为25:1。
2)将式(I)化合物半富马酸盐的粗产物在溶剂5中加热溶清，然后和溶剂6混合后，搅拌析晶，分离，干燥，得所述结晶形式的式(I)化合物半富马酸盐。
优选地，所述溶剂5选自醇类、酮类、酯类、醚类、二甲亚砜和四氢呋喃中一种或其混合；更优选地，所述溶剂5选自四氢呋喃。
优选地，所述溶剂6选自水、正庚烷、异丙醚、二氯甲烷、氯仿、甲苯、甲基环己烷中的一种或其混合；更优选地，所述溶剂6选自正庚烷。
在一些实施例中，所述溶剂5选自四氢呋喃，所述溶剂6选自正庚烷。
优选地，所述溶剂6和溶剂5的体积比≥1:1；更优选为≥1.5:1。
优选地，所述加热溶清的温度≥40℃；更优选为50℃-60℃。
优选地，所述干燥时间为≥4h；所述干燥的温度≥10℃。
在一些实施例中，所述干燥时间为8-24h；所述干燥的温度为40℃。
3)将式(I)化合物半富马酸盐在溶剂7或溶剂7和溶剂8的混合溶剂体系中加热溶清，降温搅拌，析晶，分离，干燥，得所述结晶形式的式(I)化合物半富马酸盐。
优选地，所述溶剂7选自醇类、酮类、酯类、醚类、二甲亚砜和四氢呋喃中一种或其混合；更优选地，所述溶剂7选自醇类、酮类；最优选地，所述醇类选自乙醇和甲醇中的一种或其组合；所述酮类选自丙酮。
优选地，所述溶剂8选自水、正庚烷、异丙醚、二氯甲烷、氯仿、甲苯、甲基环己烷中的一种或其混合；更优选水和正庚烷中的一种或其混合。
在一些实施例中，所述式(I)化合物半富马酸盐的粗产物在乙醇中溶清。
在一些实施例中，所述式(I)化合物在甲醇和水的混合溶剂体系中溶清，其中甲醇和水的体积比为1:0.1-0.2，优选为1:0.15。
在一些实施例中，所述式(I)化合物在丙酮和正庚烷的混合溶剂体系中溶清，其中丙酮和正庚烷的体积比为1:0.5-1，优选为1:0.8。
优选地，所述加热溶清的温度≥40℃；更优选为50℃-60℃。
优选地，所述降温搅拌的温度≤30℃；更优选为0℃-20℃。
优选地，所述干燥时间为≥4h；所述干燥的温度≥10℃。
在一些实施例中，所述干燥时间为8-24h；所述干燥的温度为40℃。
4)在式(I)化合物半富马酸盐的粗产物中加入溶剂9形成混悬液，搅拌析晶，分离，干燥，得所述结晶形式的结晶形式的式(I)化合物半富马酸盐；
优选地，所述溶剂9选自醇类、烷烃类、酯类、腈类、醚类、苯环类、酮类和水中的一种或其组合；更优选地，所述醇类选自甲醇、乙醇和异丙醇中的一种或其组合；所述烷烃类选自二氯甲烷、正庚烷、甲基环己烷中的一种或其组合；所述酯类选自乙酸乙酯、乙酸异丙酯中一种或其组合；所述腈类选自乙腈；所述醚类选自乙醚、异丙醚中的一种或其组合；所述苯环类选自甲苯；所述酮类选自丙酮和丁酮中的一种或其组合。
在一些实施例中，所述溶剂9选自单一溶剂体系，比如甲醇、水、异丙醚、二氯甲烷、异丙醇、甲基叔丁基醚、甲苯、乙腈、乙醚中的一种。
在一些实施例中，所述溶剂9为特定体积比的混合溶剂体系，比如甲醇和水的混合溶剂体系，其体积比为2：1；乙醇和水的混合溶剂体系，其体积比为3：1；丙酮和水的混合溶剂体系，其体积比为1：2；乙腈和水的混合溶剂体系，其体积比为1：4；乙酸乙酯的饱和水混合溶剂体系、乙醇和异丙醚的混合溶剂体系，其体积比为1：1；异丙醇和正庚烷的混合溶剂体系，其体积比为1：1；丁酮和甲基环己烷的混合溶剂体系，其体积比为1：5；异丙醚和乙酸乙酯的混合溶剂体系，其体积比为5：1、乙酸乙酯和二氯甲烷的混合溶剂体系，其体积比为1：5；乙酸异丙酯和甲基环己烷的混合溶剂体系，其体积比为1：5、乙醇和丙酮的混合溶剂体系，其体积比为4：1；异丙醚和乙腈的混合溶剂体系，其体积比为5：2、乙醚和甲苯的混合溶剂体系，其体积比为2：1、乙腈和甲苯的混合溶剂体系，其体积比为2：1、乙腈和乙酸乙酯的混合溶剂体系，其体积比为1：2；二氯甲烷和甲苯的混合溶剂体系，其体积比为1：10。
优选地，所述干燥时间≥4h；所述干燥的温度≥10℃。
在一些实施例中，所述干燥时间为8-24h；所述干燥的温度为40℃。
优选地，所述式(I)化合物半富马酸盐的粗产物和溶剂9的质量体积比(mg/ml)≤150:1；更优选为≤120:1。
在一些实施例中，所述式(I)化合物半富马酸盐的粗产物和溶剂9的质量体积为100:1、80:1、60:1、50:1、25:1、或20:1。
本申请的另一个方面，还在于提供一种半富马酸盐Form 1的制备方法，其包括将上述(1)-(4)任意方法中得到结晶形式的式(I)化合物半富马酸盐暴露于相对湿度≥75％的环境中足够的时间，得到式半富马酸盐Form 1。
优选地，所述暴露温度≤40℃。
优选地，所述足够的时间是指XRPD检测后确定得到半富马酸盐Form 1为止；在一些实施例中，足够的时间一般为2天以上。
本申请的另一个方面，还在于提供一种半富马酸盐Form 2的制备方法，其包括将上述(1)-(4)任意方法中得到结晶形式的式(I)化合物半富马酸盐暴露于相对湿度≤10％的环境中足够的时间，得到半富马酸盐Form 2。
优选地，所述暴露温度≥30℃。
优选地，所述足够的时间是指XRPD检测后确定得到半富马酸盐Form 2为止；在一些实施例中，足够的时间一般为2天以上。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物半富马酸盐的乙醇溶剂合物Form 3(以下简称半富马酸盐Form 3)，使用Cu-Kα辐射，所述半富马酸盐Form 3以2θ角度表示的XRPD图谱在6.8°±0.2°、9.4°±0.2°、15.7°±0.2°和15.9°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处具有特征峰。
优选地，所述半富马酸盐Form 3的XRPD图谱还在17.8°±0.2°、18.5°±0.2°、19.0°±0.2°和22.1°±0.2°2θ中的至少一处具有特征峰。
优选地，所述半富马酸盐Form 3的XRPD图谱还在23.9°±0.2°、25.1°±0.2°、26.2°±0.2°和28.6°±0.2°2θ中的至少一处具有特征峰。
非限制性地，所述Form 3的XRPD图谱在4.1°±0.2°、6.8°±0.2°、8.0°±0.2°、9.4°±0.2°、10.0°±0.2°、11.2°±0.2°、13.0°±0.2°、13.6°±0.2°、14.5°±0.2°、15.7°±0.2°、16.0°±0.2°、16.5°±0.2°、17.4°±0.2°、17.8°±0.2°、18.5°±0.2°、19.0°±0.2°、20.0°±0.2°、20.4°±0.2°、20.6°±0.2°、22.1°±0.2°、22.9°±0.2°、23.6°±0.2°、23.9°±0.2°、24.2°±0.2°、24.5°±0.2°、25.1°±0.2°、26.2°±0.2°、27.4°±0.2°、27.7°±0.2°、28.6°±0.2°、29.2°±0.2°、31.0°±0.2°、31.6°±0.2°、32.5°±0.2°、34.5°±0.2°、34.8°±0.2°、35.8°±0.2°和37.4°±0.2°2θ处具有特征峰。
优选地，所述半富马酸盐Form 3具有基本上与图11相同的XRPD图谱。
优选地，所述半富马酸盐Form 3具有基本上与图12相同的TGA图谱。
优选地，所述半富马酸盐Form 3具有基本上与图13相同的DSC图谱。
优选地，所述半富马酸盐Form 3具有基本上与图14相同的固态1H-NMR谱。
优选地，所述半富马酸盐Form 3中乙醇与式(I)化合物半富马酸盐的摩尔比为1:1。
本申请的另一个方面，还在于提供一种半富马酸盐Form 3的制备方法，所述制备方法包括以下步骤：
将结晶形式的式(I)化合物半富马酸盐在乙醇氛围中扩散结晶，得到乙醇溶剂合物Form 3。
优选地，所述扩散结晶的时间≥1天；更优选为≥5天。
在一些实施例中，所述扩散结晶的时间为6天。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物半富马酸盐的水合物Form 4(以下简称半富马酸盐Form 4)，使用Cu-Kα辐射，所述半富马酸盐Form 4以2θ角度表示的XRPD图谱在9.3°±0.2°、15.6°±0.2°、16.0°±0.2°、和21.9°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处有特征峰。
优选地，所述半富马酸盐Form 4的XRPD图谱还在17.1°±0.2°、22.8°±0.2°、24.1°±0.2°和25.0°±0.2°2θ中的至少一处具有特征峰。
优选地，所述半富马酸盐Form 4的XRPD图谱还10.0°±0.2°、9.5°±0.2°、19.0°±0.2°和20.3°±0.2°2θ中的至少一处具有特征峰。
优选地，所述半富马酸盐Form 4的XRPD图谱在3.6°±0.2°、3.8°±0.2°、6.9°±0.2°、8.1°±0.2°、8.8°±0.2°、9.3°±0.2°、9.5°±0.2°、10.0°±0.2°、11.7°±0.2°、13.7°±0.2°、15.6°±0.2°、16.0°±0.2°、16.4°±0.2°、17.1°±0.2°、17.8°±0.2°、18.6°±0.2°、19.0°±0.2°、20.3°±0.2°、20.8°±0.2°、21.9°±0.2°、22.4°±0.2°、22.8°±0.2°、23.3°±0.2°、23.6°±0.2°、24.1°±0.2°、24.4°±0.2°、25.0°±0.2°、26.6°±0.2°、27.0°±0.2°、27.7°±0.2°、28.0°±0.2°、28.7°±0.2°、31.0°±0.2°、32.8°±0.2°、34.8°±0.2°和38.6°±0.2°2θ处具有特征峰。
优选地，所述半富马酸盐Form 4具有基本上与图15相同的XRPD图谱。
优选地，所述半富马酸盐Form 4具有基本上与图16相同的TGA图谱。
优选地，所述半富马酸盐Form 4具有基本上与图17相同的DSC图谱。
优选地，所述半富马酸盐Form 4具有基本上与图18相同的DVS图谱。
优选地，所述半富马酸盐Form 4具有基本上与图19相同的固态1H-NMR谱。
本申请的另一个方面，还在于提供一种半富马酸盐Form 4的制备方法，所述制备方法包括以下方法中的任一种：
1)将式(I)化合物半富马酸盐在溶剂10中形成混悬液，搅拌析晶，分离，干燥，制得所述半富马酸盐Form 4；其中，所述溶剂10选自酯类、醇类、醚类、腈类或酮类中任一种或其组合。
优选地，所述酯类选自乙酸乙酯；所述醇类选自异丙醇和仲丁醇中的一种或其混合；所述醚类选自乙醚；所述腈类选自乙腈；所述酮类选自丙酮。
在一些实施例中，所述溶剂10为乙酸乙酯。
在一些实施例中，所述溶剂10为特定体积比的混合溶剂体系，比如丙酮和异丙醚的混合溶剂体系，其体积比为1：2；异丙醇和乙腈的混合溶剂体系，其体积比为5：2；仲丁醇和乙酸乙酯的混合溶剂体系，其体积比为5：4。
优选地，所述式(I)化合物半富马酸盐和溶剂10的质量体积比(mg/ml)≤150:1；更优选为≤120:1。
在一些实施例中，所述式(I)化合物半富马酸盐和溶剂10的质量体积为100:1、70:1或50:1。
2)将式(I)化合物半富马酸盐在溶剂11中加热溶清；挥干，得所述半富马酸盐Form 4。
优选地，所述溶剂11选自酯类、酮类溶剂中的一种或其组合；更优选地，所述酯类为乙酸乙酯；所述酮类为丙酮。
在一些实施例中，所述溶剂11为乙酸乙酯。
在一些实施例中，所述溶剂11为丙酮。
优选地，所述挥干温度≥10℃。
在一些实施例中，所述挥干温度为室温或40℃。
3)将式(I)化合物半富马酸盐在乙酸乙酯和甲苯的混合溶剂体系中加热溶清，降温搅拌，析晶，分离，干燥，制得所述半富马酸盐Form 4；其中，所述乙酸乙酯和甲苯的体积比为≥1:1。
在一些实施例中，所述乙酸乙酯和甲苯的体积比为5:1。
优选地，所述加热溶清的温度≥40℃；更优选为50℃-60℃。
优选地，所述降温搅拌的温度≤30℃；更优选为0℃-20℃。
优选地，所述干燥时间为≥4h；所述干燥的温度≥10℃。
在一些实施例中，所述干燥时间为8-24h；所述干燥的温度为40℃。
本申请所述的式(I)化合物半富马酸盐及其结晶形式具有以下有益效果：
1)稳定性良好。本申请的半富马酸盐Form 1在高温(40℃和60℃)、高湿(25℃/90％RH)和加速(40℃/75％RH)条件下放置30天，晶型基本保持不变，有关物质增加＜0.01％，具有良好的晶型稳定性和化学稳定性；半富马酸盐Form2在高温(敞口，60℃，<30％RH)条件下放置10天，晶型基本保持不变，有关物质增加＜0.05％，具有良好的晶型稳定性和化学稳定性。
2)生物利用度高。本申请的半富马酸盐Form 1在SD大鼠中具有良好的血药浓度和曲线面积以及优异的口服绝对生物利用度，在以2mg/kg的剂量口服给药时，其血药浓度和曲线面积分别是式(I)化合物游离态的1.9倍和1.8倍，其口服绝对利用度是式(I)化合物游离态的3.2倍；半富马酸盐Form 2口服绝对利用度是式(I)化合物游离态的2.7倍。
3)抑瘤效果好。本申请的半富马酸盐Form 1能显著抑制人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤的生长，肿瘤体积抑瘤率TGI(％)为75.46％，给药后的相对肿瘤增殖率T/C为23.91％；本申请的半富马酸盐Form 1能显著抑制人白血病MOLM16细胞CB-17SCID小鼠异种移植瘤的生长以及显著抑制人白血病MOLM16细胞CB-17SCID小鼠异种移植瘤的生长。
4)粒度分布好。本申请的半富马酸盐Form 1和半富马酸盐Form 2的粒径分布较好，均接近正态分布。
5)溶解度好。相对于草酸盐Form 2，本申请的半富马酸盐Form 1和半富马酸盐Form 2在pH4.5缓冲溶液中，具有更高的溶解度。
6)可压性好。相对于草酸盐Form 2制备成的片剂，半富马酸盐Form 1制备成的片剂和半富马酸盐Form 2制备成的片剂的具有更有的可压性。
7)片剂溶出好。相对于草酸盐Form 2而言，以本申请的半富马酸盐Form 1和半富马酸盐Form 2作为活性成分的片剂，具有更好的体外溶出。
8)本申请的制备方法简单，重复性高，具有可产业化前景。
本申请优选地技术方案中，所述式(I)化合物的盐选自L-酒石酸盐。
优选地，所述式(I)化合物和L-酒石酸的摩尔比为1:0.5-2；更优选为1:1。
本申请优选地技术方案中，所述式(I)化合物的盐选自L-酒石酸盐，结构式如式(III)所示:
优选地，所述式(I)化合物L-酒石酸盐基本上以结晶形式或无定型形式存在；更优选为基本上以结晶形式存在。
优选地，所述式(I)化合物L-酒石酸盐的结晶形式可以是一种或多种；具体地，可以为无水物、水合物或溶剂合物中的至少一种。
本申请的另一个方面，还在于提供一种式(I)化合物L-酒石酸盐的制备方法，其包括将式(I)化合物溶于溶剂12，然后加入L-酒石酸或L-酒石酸溶液反应得到。
本申请优选地技术方案中，所述式(I)化合物L-酒石酸盐的制备方法包括以下步骤：
步骤A:将适量式(I)化合物在溶剂12中溶解；
步骤B:将适量L-酒石酸在溶剂13中溶解；
步骤C:将上述两种溶液合并，搅拌析出固体形成混悬液；
步骤D:分离混悬液中的固体，得到式(I)化合物L-酒石酸盐；
优选地，所述溶剂12选自苯环类、烷烃类、酯类、醇类、醚类、酮类、卤素类、四氢呋喃、1,4-二氧六环中和二甲亚砜中的任一种或其组合；更优选地，所述苯环类选自甲苯；所述烷烃类选自二氯甲烷；所述醇类选自三氟乙醇、甲醇、乙醇、正丙醇、仲丁醇和正丁醇中的任一种或其组合；所述醚类选自乙醚、甲基叔丁基醚中的任一种或其组合；所述酯类选自乙酸乙酯、乙酸异丙酯和乙酸正丁酯中的任一种或其组合；所述酮类选自丙酮、丁酮中的任一种或其组合；所述卤素类选自氯仿。
在一些实施例中，所述溶剂12选自甲苯、二氯甲烷、三氟乙醇、乙醚、乙酸乙酯、乙酸异丙酯、丙酮、丁酮、氯仿、四氢呋喃和1,4-二氧六环中中的任一种或其组合。
在一些实施例中，所述溶剂12选自甲苯。
优选地，所述溶剂13选自醇类、烷烃类、酮类和四氢呋喃中的任一种或其组合；更优选地，所述醇类选自乙醇；所述烷烃类选自二氯甲烷；所述酮类选自丙酮。
在一些实施例中，所述溶剂13选自乙醇或四氢呋喃。
在一些实施例中，所述溶剂13选自丙酮/乙醇的混合溶剂或二氯甲烷/乙醇的混合溶剂。
优选地，所述步骤D的混悬液中可先加入溶剂14，再进行固体分离。
优选地，所述溶剂14选自异丙醚、正庚烷、水、异丙醇、和甲基环己烷中的任一种或其组合。
在一些实施例中，所述溶剂14选自异丙醚和正庚烷中的任一种或其组合。
优选地，所述式(I)化合物和L-酒石酸的投料摩尔比为1：1-2.5；更优选地，所述投料摩尔比为1:1。
优选地，所述式(I)化合物和溶剂12的质量体积比(mg/ml)≤300:1；更优选≤200:1；最优选≤150:1。
在一些实施例中，所述式(I)化合物和溶剂12的质量体积比(mg/ml)为100:1。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物L-酒石酸盐的Form 1(以下简称L-酒石酸盐Form 1)，使用Cu-Kα辐射，所述L-酒石酸盐Form 1以2θ角度表示的XRPD图谱在7.8°±0.2°、14.6°±0.2°、18.3°±0.2°和23.6°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处有特征峰。
优选地，所述L-酒石酸盐Form 1的XRPD图谱还在20.0°±0.2°、14.1°±0.2°、13.5°±0.2°和16.1°±0.2°2θ中的至少一处具有特征峰。
优选地，所述L-酒石酸盐Form 1的XRPD图谱还在19.8°±0.2°、13.3°±0.2°、17.0°±0.2°和15.3°±0.2°2θ中的至少一处具有特征峰。
非限制性地，所述L-酒石酸盐Form 1的XRPD图谱在3.9°±0.2°、6.1°±0.2°、7.8°±0.2°、10.2°±0.2°、11.7°±0.2°、13.3°±0.2°、13.5°±0.2°、14.1°±0.2°、14.6°±0.2°、15.3°±0.2°、16.1°±0.2°、17.0°±0.2°、17.7°±0.2°、18.3°±0.2°、18.6°±0.2°、18.8°±0.2°、19.1°±0.2°、19.8°±0.2°、20.0°±0.2°、20.3°±0.2°、22.2°±0.2°、22.6°±0.2°、23.6°±0.2°、24.2°±0.2°、24.9°±0.2°、25.4°±0.2°、26.2°±0.2°、26.7°±0.2°、28.4°±0.2°、29.0°±0.2°、30.0°±0.2°、30.8°±0.2°、31.5°±0.2°、32.7°±0.2°和32.9°±0.2°2θ处具有特征峰。
优选地，所述L-酒石酸盐Form 1具有基本上与图21相同的XRPD图谱。
优选地，所述L-酒石酸盐Form 1具有基本上与图22相同的TGA图谱。
优选地，所述L-酒石酸盐Form 1具有基本上与图23相同的DSC图谱。
优选地，所述L-酒石酸盐Form 1具有基本上与图24相同的DVS图谱。
优选地，所述L-酒石酸盐Form 1为水合物。
本申请的另一个方面，还在于提供一种L-酒石酸盐Form 1的制备方法，所述制备方法包括以下方法中的任一种：
1)步骤A:将适量式(I)化合物在溶剂15中溶解；
步骤B:将适量L-酒石酸在溶剂16中溶解；
步骤C:将上述两种溶液合并，搅拌析出固体形成混悬液；
步骤D:继续搅拌或在混悬液中加入溶剂17后继续搅拌，析晶，离心，干燥，得到L-酒石酸盐Form 1。
优选地，所述溶剂15选自苯环类；优选地，所述苯环类选自甲苯。
在一些实施例中，所述溶剂15选自甲苯。
优选地，所述溶剂16选自醇类；更优选地，所述醇类选自乙醇。
在一些实施例中，所述溶剂16选自乙醇。
优选地，所述溶剂17选自异丙醚。
优选地，所述步骤C中溶液合并的方式为将步骤B的溶液滴加到步骤A的溶液中。
优选地，所述步骤D中的干燥时间≥5h；优选≥10h。
在一些实施例中，所述干燥时间为12-24h。
在一些实施例中，所述干燥时间为16h。
优选地，所述干燥的温度为室温。
优选地，所述步骤D中的搅拌温度为低温，所述低温≤30℃；更优选为0℃-20℃；
在一些实施例中，所述步骤D中的搅拌温度为4℃。
优选地，所述式(I)化合物和L-酒石酸的投料摩尔比为1：1-2.5；更优选地，所述投料摩尔比为1:1。
优选地，所述式(I)化合物和溶剂15的质量体积比(mg/ml)≤300:1；优选为≤200:1；进一步优选为≤150:1。
在一些实施例中，所述式(I)化合物和溶剂15的质量体积比(mg/ml)为100:1。
2)取式(I)化合物L-酒石酸盐在溶剂18中形成悬浮液，搅拌析晶，得到L-酒石酸盐Form 1。
优选地，所述溶剂18选自酯类、烷烃类、醇类、醚类和水中的任一种或其组合；所述酯类选自乙酸异丙酯；所述烷烃类选自二氯甲烷、正庚烷、正己烷和甲基环己烷中的任一种或其组合；所述醇类选自甲醇和乙醇中的任一种或其组合； 所述醚类选自甲基叔丁基醚。
在一些实施例中，所述溶剂18为特定体积比的混合溶剂体系，比如乙酸异丙酯和正庚烷的混合溶剂，其体积比为1:4；或二氯甲烷和正己烷的混合溶剂，其体积比为1:4；或乙醇和正庚烷的混合溶剂，其体积比为1:4；或甲醇和水的混合溶剂，其体积比为1：2；或二氯甲烷和甲基环己烷的混合溶剂，其体积比为1:6；或甲基叔丁基醚和乙醇的混合溶剂，其体积比为4:1；或乙酸异丙酯和正己烷的混合溶剂，其体积比为1:6。
优选地，所述搅拌温度为低温、室温或高温。
优选地，所述低温≤30℃；更优选为0℃-20℃。
在一些实施例中，所述低温为4℃-8℃。
优选地，所述高温≥30℃；更优选为40℃-60℃。
在一些实施例中，所述高温为50℃。
在一些实施例中，所述式(I)化合物L-酒石酸盐和溶剂18的质量体积比(mg/mL)为40:1。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物L-酒石酸盐的Form 2(以下简称L-酒石酸盐Form 2)，使用Cu-Kα辐射，所述L-酒石酸盐Form 2具有基本上与图26相同的XRPD图谱。
优选地，所述L-酒石酸盐Form 2具有基本上与图27相同的TGA图谱。
优选地，所述L-酒石酸盐Form 2具有基本上与图28相同的DSC图谱。
优选地，所述L-酒石酸盐Form 2为水合物。
本申请的另一个方面，还在于提供一种L-酒石酸盐Form 2的制备方法，所述制备方法为将L-酒石酸盐Form 1暴露于相对湿度≥75％的环境中足够的时间，得到L-酒石酸盐Form 2。
优选地，所述暴露温度≤40℃。
在一些实施例中，所述相对湿度为75％或97％；所述暴露温度为室温。
优选地，所述足够的时间是指XRPD检测后确定得到L-酒石酸盐Form 2为止。
在一些实施例中，足够的时间为7天。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物L-酒石酸盐的Form 3(以下简称L-酒石酸盐Form 3)，使用Cu-Kα辐射，所述L-酒石酸盐Form 3具有基本上与图30相同的XRPD图谱。
优选地，所述L-酒石酸盐Form 3具有基本上与图31相同的TGA图谱。
优选地，所述L-酒石酸盐Form 3具有基本上与图32相同的DSC图谱。
优选地，所述L-酒石酸盐Form 3为水合物。
本申请的另一个方面，还在于提供一种L-酒石酸盐Form 3的制备方法，所述制备方法包括：
将L-酒石酸盐Form 1在溶剂19中形成悬浮液，低温搅拌析出，分离晶体，得L-酒石酸盐Form 3；其中溶剂19为二甲基四氢呋喃和烷烃类溶剂的混合溶剂。
在一些实施例中，所述溶剂19为二甲基四氢呋喃和甲基环己烷的混合溶剂，其体积比为1：4。
优选地，所述低温搅拌的温度≤30℃；优选为0℃-20℃。
在一些实施例中，所述低温搅拌的温度为4℃-8℃。
优选地，所述搅拌时间≥0.5天；更优选为1-10天。
在一些实施例中，所述搅拌时间为3天。
优选地，所述L-酒石酸盐Form 1和溶剂19的质量体积比(mg/mL)为40:1。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物L-酒石酸盐的Form 4(以下简称L-酒石酸盐Form 4)，使用Cu-Kα辐射，所述L-酒石酸盐Form 4以2θ角度表示的XRPD图谱在3.6°±0.2°、11.0°±0.2°和14.3°±0.2°中的至少一处具有特征峰；优选为二处或三处有特征峰。
优选地，所述L-酒石酸盐Form 4的XRPD图谱还在6.6°±0.2°、7.3°±0.2°、8.4°±0.2°、17.7°±0.2°和18.0°±0.2°2θ中的至少一处具有特征峰。
优选地，所述L-酒石酸盐Form 4的XRPD图谱还在13.2°±0.2°、13.8°±0.2°、18.5°±0.2°19.9°±0.2°和21.8°±0.2°2θ中的至少一处具有特征峰。
非限制性地，所述L-酒石酸盐Form 4的XRPD图谱在3.6°±0.2°、6.6°±0.2°、7.3°±0.2°、8.4°±0.2°、11.0°±0.2°、12.5°±0.2°、13.2°±0.2°、13.8°±0.2°、14.0°±0.2°、14.3°±0.2°、14.7°±0.2°、15.6°±0.2°、16.0°±0.2°、16.9°±0.2°、17.5°±0.2°、17.7°±0.2°、18.0°±0.2°、18.5°±0.2°、18.8°±0.2°、19.2°±0.2°、19.5°±0.2°、19.9°±0.2°、21.8°±0.2°、22.1°±0.2°、22.3°±0.2°、22.9°±0.2°、23.3°±0.2°、24.9°±0.2°、25.0°±0.2°、25.4°±0.2°、25.6°±0.2°、25.9°±0.2°、26.2°±0.2°、26.7°±0.2°、27.2°±0.2°、28.5°±0.2°、29.9°±0.2°、30.7°±0.2°、30.9°±0.2°、31.8°±0.2°、32.2°±0.2°、32.5°±0.2°和36.0°±0.2°2θ处具有特征峰。
优选地，所述L-酒石酸盐Form 4具有基本上与图34相同的XRPD图谱。
优选地，所述L-酒石酸盐Form 4具有基本上与图35相同的TGA图谱。
优选地，所述L-酒石酸盐Form 4具有基本上与图36相同的DSC图谱。
在一些实施例中，所述L-酒石酸盐Form 4为无水物或通道水合物。
本申请的另一个方面，还在于提供一种L-酒石酸盐Form 4的制备方法，所述制备方法包括以下方法中的任一种：
1)将L-酒石酸盐Form 1在溶剂20中溶清，然后缓慢挥发析晶，分离晶体，得到L-酒石酸盐Form 4。
优选地，所述溶剂20选自酮类中、酯类、醚类和苯环类的任一种或其混合；更优选地，所述酮类选自丙酮和甲基异丁酮中的任一种或其组合；所述酯类选自乙酸乙酯和乙酸异丙酯中的任一种或其组合；所述醚类选自甲基叔丁基醚；所述苯环类选自甲苯。
在一些实施例中，所述溶剂20为单一溶剂体系，比如乙酸异丙酯。
在一些实施例中，所述溶剂20为特定体积比的混合溶剂体系，比如丙酮和甲基叔丁基醚的混合溶剂，其体积比为1:1；或者乙酸乙酯和甲苯的混合溶剂体系，其体积比为2:1。
优选地，所述挥发的时间≥0.5天；在一些实施例中，所述挥发时间为1天。
优选地，所述缓慢挥发为小孔挥发。
优选地，所述缓慢挥发的温度为室温。
2)将L-酒石酸盐Form 1在溶剂21中形成悬浮液，加入L-酒石酸盐Form 4晶种，室温搅拌析出，分离晶体，得到L-酒石酸盐Form 4。
优选地，所述溶剂21为酮类、烷烃类、酯类和醚类中的任一种或其组合；更优选地，所述酮类选自丙酮和甲基异丁酮中的任一种或其组合；所述烷烃类选自正庚烷；所述酯类选自乙酸乙酯；所述醚类选自甲基叔丁基醚。
在一些实施例中，所述溶剂21为特定体积比的混合溶剂体系，比如丙酮和正庚烷的混合溶剂，其体积比为1:6；或乙酸乙酯和正庚烷的混合溶剂，其体积比为1:6；或乙酸异丙酯和正庚烷的混合溶剂，其体积比为1:4，或丙酮和甲基叔丁基醚的混合溶剂，其体积比为1:4。
在一些实施例中，所述L-酒石酸盐Form 1和溶剂21的质量体积比(mg/mL)为40:1。
3)将适量式(I)化合物和适量L-酒石酸在溶剂22中形成悬浮液，加入L-酒石酸盐Form 4晶种，搅拌析出，分离晶体，得L-酒石酸盐Form 4。
优选地，所述溶剂22选自为酮类、烷烃类、酯类和醚类中的任一种或其组合；更优选地，所述酮类选自丙酮和甲基异丁酮中的任一种或其组合；所述烷烃类选自正庚烷；所述酯类选自乙酸乙酯；所述醚类选自甲基叔丁基醚。
在一些实施例中，所述溶剂22为特定体积比的混合溶剂体系，比如丙酮和正庚烷的混合溶剂，其体积比为1:6。
在一些实施例中，所述式(I)化合物和L-酒石酸的投料摩尔比为1:1。
在一些实施例中，所述式(I)化合物和溶剂22的质量体积比(mg/mL)为30:1。
4)取式(I)化合物L-酒石酸盐，在溶剂23中形成悬浮液，搅拌析晶，得到L-酒石酸盐Form 4。
优选地，所述溶剂23选自烷烃类、酮类和醚类中的任一种或其组合；更优选地，所述烷烃类选自正庚烷；所述酮类选自丁酮；所述醚类选自甲基叔丁基醚。
在一些实施例中，所述溶剂23为甲基叔丁基醚。
在一些实施例中，所述溶剂23为特定体积比的混合溶剂体系，比如丁酮和正庚烷的混合溶剂，其体积比为1:6。
优选地，所述搅拌温度为低温或室温。
优选地，所述低温≤30℃；更优选为0℃-20℃。
在一些实施例中，所述低温为4℃-8℃。
在一些实施例中，所述式(I)化合物L-酒石酸盐和溶剂23的质量体积比(mg/mL)为40:1。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物L-酒石酸盐的Form 5(以下简称L-酒石酸盐Form 5)，使用Cu-Kα辐射，所述L-酒石酸盐 Form 5具有基本上与图38相同的XRPD图谱。
本申请的另一个方面，还在于提供一种L-酒石酸盐Form 5的制备方法，所述制备方法为将L-酒石酸盐Form 1在异丙醇和甲基叔丁基醚的混合溶剂中形成悬浮液，搅拌析出，分离晶体，得到L-酒石酸盐Form 5。
在一些实施例中，所述异丙醇和甲基叔丁基醚的体积比为1:1。
在一些实施例中，所述搅拌为室温搅拌。
在一些实施例中，所述搅拌时间为3天。
在一些实施例中，所述L-酒石酸盐Form 1和混合溶剂的质量体积比(mg/mL)为40:1。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物L-酒石酸盐Form 6(以下简称L-酒石酸盐Form 6)，使用Cu-Kα辐射，所述L-酒石酸Form6具有基本上与图39相同的XRPD图谱。
本申请的另一个方面，还在于提供一种L-酒石酸盐Form 6的制备方法，所述制备方法为将L-酒石酸盐Form 1加热至70℃，并保持10分钟，得到L-酒石酸盐Form 6。
本申请的另一个方面，还在于提供一种无定型形式的式(I)化合物L-酒石酸盐(以下简称L-酒石酸盐无定型)，使用Cu-Kα辐射，所述L-酒石酸盐无定型以2θ表示的XRPD图谱中无尖锐的衍射峰。
优选地，所述L-酒石酸盐无定型具有基本上与40相同的XRPD图谱。
优选地，所述L-酒石酸盐无定型具有基本上与41相同的TGA图谱。
优选地，所述L-酒石酸盐无定型具有基本上与42相同的DSC图谱。
优选地，所述L-酒石酸盐无定型具有基本上与图43相同的DVS图谱。
本申请的另一个方面，还在于提供一种L-酒石酸盐无定型的制备方法，所述制备方法为将式(I)化合物L-酒石酸盐在二氯甲烷中溶解，过滤，将滤液高温浓缩，得L-酒石酸盐无定型。
优选地，所述浓缩为减压浓缩。
优选地，所述高温为40-70℃。
在一些实施例中，所述高温为60℃。
在一些实施例中，所述式(I)化合物L-酒石酸盐和二氯甲烷的质量体积比(mg/ml)为10:1。
本申请所述的式(I)化合物L-酒石酸盐及其结晶形式具有以下有益效果：
1)稳定性良好。本申请的L-酒石酸盐Form 1在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置1周，晶型和纯度基本保持不变，在高温(60℃,<30％RH)条件下放置10天，其晶型基本保持不变，有关物质增加＜0.05％，具有良好的晶型稳定性和化学稳定性；本申请的L-酒石酸盐Form 4在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置1周，晶型和纯度基本保持不变，分别在40℃、50℃和55℃条件下放置30天，其晶型基本保持不变，具有良好的晶型稳定性和化学稳定性。
2)生物利用度高。本申请的L-酒石酸盐Form 1在SD大鼠中具有良好的血药浓度和曲线面积以及优异的口服绝对生物利用度，在以10mg/kg的剂量口服给药时，其血药浓度和曲线面积分别是式(I)化合物游离态的1.5倍和1.8倍，其口服绝对利用度是式(I)化合物游离态的2.6倍；本申请的L-酒石酸盐Form 4在SD大鼠中具有良好的血药浓度和曲线面积以及优异的口服绝对生物利用度，在以2mg/kg的剂量口服给药时，其血药浓度和曲线面积分别是式(I)化合物游离态的3倍和2.7倍，其口服绝对利用度是式(I)化合物游离态的3.8倍。
3)抑瘤效果好。本申请的L-酒石酸盐Form 1在NOD-SCID免疫缺陷鼠Mol m16皮下肿瘤模型中具有显著优于溶媒的抑瘤效果，且同等剂量下，其抑瘤效果优于式(I)化合物游离态；本申请的L-酒石酸盐Form 4能显著抑制人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤的生长，肿瘤体积抑瘤率TGI(％)为79.43％，给药后的相对肿瘤增殖率T/C为20.37％；本申请的L-酒石酸盐Form 4显著抑制人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤的生长和人非小细胞肺癌NCI-H460细胞Balb/c裸鼠异种移植瘤的生长。
4)流动性好。本申请的L-酒石酸盐Form 4和L-酒石酸盐Form 1的流动性好，且均优于草酸盐Form 2。
5)片剂溶出好。
6)本申请的制备方法简单，重复性高，具有可产业化前景。
本申请优选地技术方案中，所述式(I)化合物的盐选自盐酸盐，结构式如式(IV)所示：
优选地，所述式(I)化合物的盐酸盐基本上以结晶形式或无定型形式存在；更优选为基本上以结晶形式存在。
优选地，所述式(I)化合物盐酸盐的结晶形式可以是一种或多种；具体地，可以为无水物、水合物或溶剂合物中的至少一种。
本申请的另一个方面，在于提供一种结晶形式的式(I)化合物盐酸盐的Form1(以下简称盐酸盐Form 1)，使用Cu-Kα辐射，所述盐酸盐Form 1以2θ角度表示的XRPD图谱13.5°±0.2°、15.4°±0.2°、15.7°±0.2°、32.6°±0.2°和35.3°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处或五处有特征峰。
优选地，所述盐酸盐Form 1具有基本上与图45相同的XRPD图谱。
优选地，所述盐酸盐Form 1具有基本上与图46相同的TGA图谱。
优选地，所述盐酸盐Form 1具有基本上与图47相同的DSC图谱。
本申请的另一个方面，还在于提供一种盐酸盐Form 1的制备方法，所述制备方法包括以下：
步骤A:将适量式(I)化合物在酯类溶剂中溶解；
步骤B:将适量盐酸在醇类溶剂中溶解；
步骤C:合并两种溶液，搅拌析出固体形成混悬液；
步骤D:将混悬液搅拌析晶，离心，干燥，得式盐酸盐Form 1。
在一些实施例中，所述酯类溶剂选自乙酸丁酯。
在一些实施例中，所述醇类溶剂选自甲醇。
在一些实施例中，所述步骤D中的搅拌温度为4℃。
在一些实施例中，所述干燥时间为16h。
在一些实施例中，所述干燥的温度为室温。
在一些实施例中，所述式(I)化合物和盐酸的投料摩尔比为1:1.2。
在一些实施例中，所述式(I)化合物和酯类溶剂的质量体积比(mg/ml)为100:1。
本申请优选地技术方案中，所述式(I)化合物的盐选自甲磺酸盐，结构式如式(V)所示：
优选地，所述式(I)化合物的甲磺酸盐基本上以结晶形式或无定型形式存在；更优选为基本上以结晶形式存在。
优选地，所述式(I)化合物甲磺酸盐的结晶形式可以是一种或多种；具体地，可以为无水物、水合物或溶剂合物中的至少一种。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物甲磺酸盐的Form 1(以下简称甲磺酸盐Form 1)，使用Cu-Kα辐射，所述甲磺酸盐Form 1具有基本上与图49相同的XRPD图谱。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物甲磺酸盐的Form 2(以下简称甲磺酸盐Form 2)，使用Cu-Kα辐射，所述甲磺酸盐Form 2以2θ角度表示的XRPD图谱在6.5°±0.2°、13.8°±0.2°、15.2°±0.2°和17.0°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处有特征峰。
优选地，所述甲磺酸盐Form 2的XRPD图谱还在3.9°±0.2°、4.6°±0.2°、4.9°±0.2°、10.5°±0.2°、10.7°±0.2°、12.8°±0.2°、13.0°±0.2°、16.3°±0.2°、16.7°±0.2°、17.4°±0.2°、18.3°±0.2°、18.8°±0.2°、19.6°±0.2°、20.6°±0.2°、21.2°±0.2°、22.4°±0.2°、23.0°±0.2°、23.9°±0.2°、25.6°±0.2°、25.8°±0.2°、26.6°±0.2°、29.6°±0.2°和31.4°±0.2°2θ中的至少一处有特征峰。
优选地，所述甲磺酸盐Form 2具有基本上与图50相同的XRPD图谱。
优选地，所述甲磺酸盐Form 2具有基本上与图51相同的TGA图谱。
优选地，所述甲磺酸盐Form 2具有基本上与图52相同的DSC图谱。
本申请的另一个方面，还在于提供一种甲磺酸盐Form 2的制备方法，所述制备方法包括：
将式(I)化合物和甲磺酸分别在苯环类溶剂中溶解，将两种溶液混合，低温搅拌，析出，加入醚类溶剂，低温搅拌，离心，干燥，得甲磺酸盐Form 2。
在一些实施例中，所述苯环类溶剂为甲苯。
在一些实施例中，所述甲磺酸和甲苯的质量体积比(mg/ml)为72.5:1。
在一些实施例中，所述式(I)化合物和甲苯的质量体积比(mg/ml)为100:1。
在一些实施例中，所述式(I)化合物和甲磺酸的投料摩尔比为1:1.1。
在一些实施例中，所述醚类溶剂为异丙醚。
在一些实施例中，所述低温搅拌的温度为4℃。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物甲磺酸盐的Form 3(以下简称甲磺酸盐Form 3)，使用Cu-Kα辐射，所述甲磺酸盐Form 3具有基本上与图54相同的XRPD图谱。
本申请优选地技术方案中，所述式(I)化合物的盐选自草酸盐，结构式如式(VI)所示:
优选地，所述式(I)化合物草酸盐基本上以结晶形式或无定型形式存在。
优选地，所述式(I)化合物草酸盐的结晶形式可以是一种或多种；具体地，可以为无水物、水合物或溶剂合物中的至少一种。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物草酸盐的Form 1(以下简称草酸盐Form 1),使用Cu-Kα辐射，所述草酸盐Form 1具有基本上与图55相同的XRPD图谱。
本申请的另一个方面，还在于提供一种结晶形式的式(I)化合物草酸盐的Form 2(以下简称草酸盐Form 2)，使用Cu-Kα辐射，所述草酸盐Form 2以2θ角度表示的XRPD图谱在15.5°±0.2°、17.2°±0.2°和23.8°±0.2°中的至少一处具有特征峰；优选为二处或三处有特征峰。
优选地，所述草酸盐Form 2的XRPD图谱还在13.9°±0.2°、16.9°±0.2°、19.8°±0.2°和16.0°±0.2°2θ中的至少一处具有特征峰。
优选地，所述草酸盐Form 2具有基本上与图56相同的XRPD图谱。
本申请的另一个方面，还在于提供一种草酸盐Form 2的制备方法，所述制备方法包括：将式(I)化合物和草酸分别在酮类溶剂中溶解，将两种溶液混合，室温搅拌，析出，加入醚类溶剂，搅拌，离心，干燥，得草酸盐Form 2。
在一些实施例中，所述酮类溶剂为丙酮。
在一些实施例中，所述醚类溶剂为异丙醚。
在一些实施例中，所述式(I)化合物和丙酮的质量体积比(mg/ml)＞100:1。
在一些实施例中，所述式(I)化合物和草酸的投料摩尔比为1:1-1.5。
在一些实施例中，所述低温搅拌的温度为4℃。
本申请的另一个方面，还在于提供一种药物组合物，所述药物组合物包含治疗有效量的式(I)化合物的盐或其晶型，以及至少一种药学上可接受的载体。
本申请的另一个方面，还在于提供一种上述药物组合物制备成的制剂，所述制剂形式包括但不限于口服固体制剂、注射剂、外用制剂。
本申请优选技术方案中，所述制剂形式为片剂、胶囊，粉针剂，粉剂、糖浆、溶液状、悬浮液、气雾剂或栓剂。
本申请的各种剂型可按照药学领域的常规制备方法制备。
本申请优选技术方案中，所述药学上可接受的载体为本领域制剂中常用的辅料，包括但不限于粘合剂、表面活性剂、稀释剂、抗粘附剂、亲水性或疏水性高分子聚合物、安定剂或稳定剂、崩解剂、抗氧化剂、消泡剂、填充剂、助流剂/润滑剂、吸附剂、防腐剂、增塑剂、甜味剂中的任一种及其两种及以上的混合物。
本申请优选技术方案中，当所述制剂为口服固体制剂时，所述填充剂或稀释剂选自乳糖、D-甘露糖醇、微晶纤维素、淀粉、预胶化淀粉、硫酸钙、磷酸氢钙、碳酸钙中的任一种或其组合；所述崩解剂选自羧甲基淀粉钠、羟甲基纤维素钠、交联羧甲基纤维素钠、低取代羟丙基纤维素、交联聚乙烯吡咯烷酮中的任一种或其组合；所述润滑剂/助流剂选自硬脂酸镁、滑石粉、微粉硅胶中的一种或其组合。
本申请优选技术方案中，所述药物组合物还可以包含一种或多种pH调整剂或缓冲剂，举例来说：酸，例如乙酸、硼酸、柠檬酸、富马酸、马来酸、酒石酸、苹果酸、乳酸、磷酸、盐酸的任一种或其组合；或者碱，例如氢氧化钠、磷酸钠、硼酸钠、柠檬酸钠、乙酸钠、乳酸钠、三羟甲基氨基甲烷的任一种或其组合物；或者缓冲剂，例如柠檬酸盐/葡萄糖、碳酸氢钠、氯化铵和类似物；用作碱的此类缓冲剂可具有除钠以外的平衡离子，例如钾、镁、钙、铵和其它平衡离子；以及其他将组分的pH维持在可接受范围内所需的量，包含此类酸、碱和缓冲剂的溶液或固体。
本申请的另一个方面，还在于提供一种上述式(I)化合物的盐或其晶型、或其药物组合物在制备预防和/或治疗STAT3双功能磷酸化位点活化引起或调控的疾病的药物中的应用。
本申请优选地技术方案中，所述应用为抑制癌细胞的增殖、生长、迁移、浸润、克隆形成和转移，促进癌细胞的凋亡，和/或延长肿瘤患者的生存期。
本申请优选地技术方案中，所述疾病包括但不限于恶性肿瘤的例子包括星形神经胶质瘤、恶性成神经管细胞瘤、胚细胞肿瘤，颅咽管瘤、室管膜瘤等小儿脑肿瘤；神经胶质瘤、脑膜瘤、脑下垂体腺瘤，神经鞘瘤等成人脑肿瘤；上颌窦癌、咽喉癌(鼻咽癌、中咽喉癌、喉咽癌)、喉癌、口腔癌、唇癌、舌癌、腮腺癌等头颈部癌；小细胞肺癌、非小细胞肺癌、胸腺瘤、间皮瘤等胸部癌及肿瘤；食道癌、肝癌、原发性肝癌、胆囊癌、胆管癌、胃癌、大肠癌、结肠癌、直肠癌、肛门癌、胰癌、胰内分泌肿瘤等消化道癌及肿瘤；阴茎癌、肾盂·输尿管癌、肾细胞癌、睾丸肿瘤、前列腺癌、膀胱癌、肾母细胞瘤、泌尿道上皮癌等泌尿器癌及肿瘤；外阴癌、子宫颈癌、子宫体癌、子宫内膜癌、子宫肉瘤、绒毛膜癌、阴道癌、乳腺癌、卵巢癌、卵巢胚细胞肿瘤等妇科癌及肿瘤；成人及小儿软组织肉瘤；骨肉瘤、尤因瘤等骨肿瘤；肾上腺皮质癌、甲状腺癌等内分泌组织癌及肿瘤；恶性淋巴瘤、非何杰金氏淋巴瘤、何杰金氏病、多发性骨髓瘤、浆细胞性肿瘤、急性骨髓性白血病、急性淋巴性白血病、成人T细胞白血病淋巴瘤、慢性骨髓性白血病、慢性淋巴性白血病等恶性淋巴瘤及白血病；慢性骨髓增生性疾病、恶性黑素瘤、鳞状细胞癌、基底细胞癌、蕈样肉芽肿病等皮肤癌及肿瘤；上述肿瘤及癌的转移病灶等。
优选地，所述疾病选自胰腺癌Capan-2、PANC-1、MIAPACA-2、BXPC3、SW1990、CFPAC-1、Pan02、ASPC-1，胃癌MKN45、BGC823、MGC803，肝癌Huh7、PLC/PRF-5、HepG2、SK-hep1、SMMC7721、HCCC9810、肺癌HCC827、A549、H460、H23、H1299、H522、H1975、PC-9，前列腺癌DU145、LNCAP，结肠癌HCT8、HT29、HCT-116。
本申请的另一个方面，还在于提供一种上述式(I)化合物的盐或其晶型、或其药物组合物在制备抑制STAT3的磷酸化、抑制STAT3的转录活性和线粒体的氧化磷酸化的抑制剂中的应用。
本申请的另一个方面，还在于提供一种预防和/或治疗与STAT3双功能磷酸化位点活化有关的疾病的方法，其包括向有需要的个体中施于有效量的上述式(I)化合物的盐或其晶型、或其药物组合物。
本申请优选地技术方案中，所述疾病包括但不限于恶性肿瘤的例子包括星形神经胶质瘤、恶性成神经管细胞瘤、胚细胞肿瘤，颅咽管瘤、室管膜瘤等小儿脑肿瘤；神经胶质瘤、脑膜瘤、脑下垂体腺瘤，神经鞘瘤等成人脑肿瘤；上颌窦癌、咽喉癌(鼻咽癌、中咽喉癌、喉咽癌)、喉癌、口腔癌、唇癌、舌癌、腮腺癌等头颈部癌；小细胞肺癌、非小细胞肺癌、胸腺瘤、间皮瘤等胸部癌及肿瘤；食道癌、肝癌、原发性肝癌、胆囊癌、胆管癌、胃癌、大肠癌、结肠癌、直肠癌、肛门癌、胰癌、胰内分泌肿瘤等消化道癌及肿瘤；阴茎癌、肾盂·输尿管癌、肾细胞癌、睾丸肿瘤、前列腺癌、膀胱癌、肾母细胞瘤、泌尿道上皮癌等泌尿器癌及肿瘤；外阴癌、子宫颈癌、子宫体癌、子宫内膜癌、子宫肉瘤、绒毛膜癌、阴道癌、乳腺癌、卵巢癌、卵巢胚细胞肿瘤等妇科癌及肿瘤；成人及小儿软组织肉瘤；骨肉瘤、尤因瘤等骨肿瘤；肾上腺皮质癌、甲状腺癌等内分泌组织癌及肿瘤；恶性淋巴瘤、非何杰金氏淋巴瘤、何杰金氏病、多发性骨髓瘤、浆细胞性肿瘤、急性骨髓性白血病、急性淋巴性白血病、成人T细胞白血病淋巴瘤、慢性骨髓性白血病、慢性淋巴性白血病等恶性淋巴瘤及白血病；慢性骨髓增生性疾病、恶性黑素瘤、鳞状细胞癌、基底细胞癌、蕈样肉芽肿病等皮肤癌及肿瘤；上述肿瘤及癌的转移病灶等。
优选地，所述疾病选自胰腺癌Capan-2、PANC-1、MIAPACA-2、BXPC3、SW1990、CFPAC-1、Pan02、ASPC-1、胃癌MKN45、BGC823、MGC803，肝癌Huh7、PLC/PRF-5、HepG2、SK-hep1、SMMC7721、HCCC9810，肺癌HCC827、A549、H460、H23、H1299、H522、H1975、PC-9，前列腺癌DU145、LNCAP，结肠癌HCT8、HT29和HCT-116。
本申请的另一个方面，还在于提供一种上述式(I)化合物的盐或其晶型其药物组合物与其他药物的联合应用。
优选地，所述其它药物是用于预防和/或治疗癌症、肾纤维化、肺纤维化、类风湿性关节炎、银屑病、红斑狼疮、炎性肺病和炎性肠病等的药物。
除非特殊注明：
实验操作温度一般指室温，“室温”是指10℃～30℃的温度。
当由本领域技术人员考虑时，术语“约”意指落入可接受的平均值误差标准内的值。
本申请中，“晶体”或“晶型”指的是被所示的X-射线粉末衍射图表征所证实的。本领域技术人员公知，其中的实验误差取决于仪器条件、样品的准备和样品的纯度，特别是X-射线粉末衍射图通常会随着仪器的条件而有所改变。需要特别指出的是，X-射线粉末衍射图的相对强度也可能随着实验条件的变化而变化，所以峰强度的顺序不能作为唯一或决定性因素。另外，峰角度的实验误差通常在5％或更少，这些角度的误差也应该被考虑进去，通常允许有±0.2°的误差。另外，由于样品高度等实验因素的影响，会造成峰角度的整体偏移，通常允许一定的偏移。因而，本领域技术人员可以理解的是，任何具有和本申请晶型的X-射线粉末衍射图谱中的特征峰相同或相似的晶型均属于本申请的范畴。“搅拌”，可以采用本领域的常规方法，例如搅拌方式包括磁力搅拌、机械搅拌，搅拌速度为50～1800转/分，优选为300～900转/分。
“分离”可以采用本领域的常规方法，例如离心或过滤。优选减压过滤，一般是以小于大气压的压力进行抽滤，优选压力小于0.09MPa。
干燥”，可以采用本领域的常规技术完成，例如常温干燥、鼓风干燥或减压干燥；可以减压或常压，优选压力小于0.09MPa。干燥仪器和方法不受限制，可以是通风橱、鼓风烘箱、喷雾干燥器、流化床干燥或真空烘箱；可以在减压或不减压下进行，优选为压力小于0.09Mpa。
如无特殊说明，本申请涉及的比例，液体与固体之间，为质量体积比，液体与液体之间，为体积比。
本申请中，基本上以结晶形式存在，是指X射线粉末衍射数据所测定的结晶度大于约20％，更优选大于约60％、甚至大于约80％，更优选大于约90％。
附图说明
图1为实施例2中半富马酸盐Form 1的XRPD图谱；
图2为实施例2中半富马酸盐Form 1的TGA图谱；
图3为实施例2中半富马酸盐Form 1的DSC图谱；
图4为实施例2中半富马酸盐Form 1的DVS图谱；
图5为实施例2中半富马酸盐Form 1的固态1H-NMR谱；
图6为实施例2中半富马酸盐Form 2的XRPD图谱；
图7为实施例2中半富马酸盐Form 2的TGA图谱；
图8为实施例2中半富马酸盐Form 2的DSC图谱；
图9为实施例2中半富马酸盐Form 2的DVS图谱；
图10为实施例2中半富马酸盐Form 2的固态1H-NMR谱；
图11为实施例35中半富马酸盐Form 3的XRPD图谱；
图12为实施例35中半富马酸盐Form 3的TGA图谱；
图13为实施例35中半富马酸盐Form 3的DSC图谱；
图14为实施例35中半富马酸盐Form 3的固态1H-NMR谱；
图15为实施例36中半富马酸盐Form 4的XRPD图谱；
图16为实施例36中半富马酸盐Form 4的TGA图谱；
图17为实施例36中半富马酸盐Form 4的DSC图谱；
图18为实施例36中半富马酸盐Form 4的DVS图谱；
图19为实施例36中半富马酸盐Form 4的固态1H-NMR谱；
图20为实施例44中半富马酸盐Form 1的单晶模拟图谱；
图21为实施例46中L-酒石酸盐Form 1的XRPD图谱；
图22为实施例46中L-酒石酸盐Form 1的TGA图谱；
图23为实施例46中L-酒石酸盐Form 1的DSC图谱；
图24为实施例46中L-酒石酸盐Form 1的DVS图谱；
图25为实施例46中L-酒石酸盐Form 1的固态1H-NMR谱；
图26为实施例57中L-酒石酸盐Form 2的XRPD图谱；
图27为实施例57中L-酒石酸盐Form 2的TGA图谱；
图28为实施例57中L-酒石酸盐Form 2的DSC图谱；
图29为实施例57中L-酒石酸盐Form 2的固态1H-NMR谱；
图30为实施例58中L-酒石酸盐Form 3的XRPD图谱；
图31为实施例58中L-酒石酸盐Form 3的TGA图谱；
图32为实施例58中L-酒石酸盐Form 3的DSC图谱；
图33为实施例58中L-酒石酸盐Form 3的固态1H-NMR谱；
图34为实施例59中L-酒石酸盐Form 4的XRPD图谱；
图35为实施例59中L-酒石酸盐Form 4的TGA图谱；
图36为实施例59中L-酒石酸盐Form 4的DSC图谱；
图37为实施例59中L-酒石酸盐Form 4的固态1H-NMR谱；
图38为实施例74中L-酒石酸盐Form 5的XRPD图谱；
图39为实施例75中L-酒石酸盐Form 6的XRPD图谱；
图40为实施例76中L-酒石酸盐无定型的XRPD图谱；
图41为实施例76中L-酒石酸盐无定型的TGA图谱；
图42为实施例76中L-酒石酸盐无定型的DSC图谱；
图43为实施例76中L-酒石酸盐无定型的DVS图谱；
图44为实施例76中L-酒石酸盐无定型的固态1H-NMR谱；
图45为实施例77中盐酸盐Form 1的XRPD图谱；
图46为实施例77中盐酸盐Form 1的TGA图谱；
图47为实施例77中盐酸盐Form 1的DSC图谱；
图48为实施例77中盐酸盐Form 1的DVS图谱；
图49为实施例78中甲磺酸盐Form 1的XRPD图谱；
图50为实施例79中甲磺酸盐Form 2的XRPD图谱；
图51为实施例79中甲磺酸盐Form 2的TGA图谱；
图52为实施例79中甲磺酸盐Form 2的DSC图谱；
图53为实施例79中甲磺酸盐Form 2的DVS图谱；
图54为实施例80中甲磺酸盐Form 3的XRPD图谱；
图55为实施例81中草酸盐Form 1的XRPD图谱；
图56为实施例82中草酸盐Form 2的XRPD图谱；
图57为实施例82中草酸盐Form 2的TGA图谱；
图58为实施例82中草酸盐Form 2的DSC图谱；
图59为实施例82中草酸盐Form 2的DVS图谱；
图60为实验例1中半富马酸盐Form 1在高温(40℃和60℃)条件下放置前后的XRPD叠图；
图61为实验例1中半富马酸盐Form 1在高湿(25℃/90％RH)、加速(40℃/75％RH)条件下放置前后的XRPD叠图；
图62为实验例1中半富马酸盐Form 2在高温(敞口，60℃，<30％RH)条件下放置前后的XRPD叠图；
图63为实验例1中L-酒石酸盐Form 1在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置前后的XRPD叠图；
图64为实验例1中L-酒石酸盐Form 1在高温(60℃,<30％RH)条件下放置前后的XRPD叠图；
图65为实验例1中L-酒石酸盐Form 4在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置前后的XRPD叠图；
图66为实验例1中L-酒石酸盐Form 4在40℃条件下放置前后的XRPD叠图；
图67为实验例1中L-酒石酸盐Form 4在50℃条件下放置前后的XRPD叠图；
图68为实验例1中L-酒石酸盐Form 4在55℃条件下放置前后的XRPD叠图；
图69为实验例2中半富马酸盐Form 1的PSD图；
图70为实验例2中半富马酸盐Form 2的PSD图；
图71为实验例2中草酸盐Form 2的PSD图。
具体实施方式
实施例
下面结合附图和实施例对本申请的技术方案进行详细描述，但并不因此将本申请限制在所述的实施例范围之中。
在本申请中，X-射线粉末衍射(XRPD)数据采自于BrukerD8 Advance diffractometer；参数如下：Cu靶；波长为电流电压：40KV，40mA；角度范围：3～40°2θ。
在本申请中，热重分析(TGA)数据采自于TA Instruments Q500TGA；参数如下：模式：高分辨模式；升温速率：10℃/min；保护气体：N2；样品盘：铂金坩埚。
在本申请中，差热分析(DSC)数据采自于TA Instruments Q200 DSC；参数如下：升温速率：10℃/min；保护气体：N2；样品盘：加盖的铝坩埚。
在本申请中，动态水份吸附分析(DVS)数据和等温吸附分析数据采自于TA Instruments Q5000 SA；参数如下：温度：25℃；相对湿度范围：0％RH-80％RH；dm/dt＝0.001％/min；平衡时间：90min；保护气体：N2；样品盘：铂金坩埚。
在本申请中，固态1H-NMR谱数据采集于WNMR-I 400。
在本申请中，真空干燥采用真空干燥箱，型号为D2F-6020。
在本申请中，分析天平型号为BT125D。
本申请中，稳定性研究中高效液相色谱仪(HPLC)检测参数如下：色谱仪：Ultimate3000，色谱柱：C18 3/5μm(150*4.6mm)，柱温：35℃，流速：1.0mL/min，检测波长：254nm，流动相：流动相A：水，流动相B：甲醇，流动相运行条件如下：

在本申请中，晶型的溶解度和片剂溶出研究中高效液相色谱仪(HPLC)检测参数如下：色谱仪：Ultimate U3000，色谱柱：C18 150×4.6mm,3.5μm，柱温：40℃，流速：1.0mL/min，检测波长：220nm，流动相：流动相A：0.05％的三氟乙酸水溶液，流动相B：乙腈溶液，流动相运行条件如下：
下列实施例中未注明具体条件的实验方法，按照常规方法和条件，或按照商品说明书选择。本申请所用试剂和原料除了特殊说明外，均市售可得。
本申请中，起始物料式(I)化合物可通过CN112300145A中提到的方法制备得到。
实施例1：式(I)化合物盐型的制备
1)以甲醇作为溶剂1进行成盐反应
成盐步骤：取式(I)化合物约50mg,60℃下溶于2.0mL甲醇中，分别加入不同的对离子酸溶液(摩尔比为碱:酸＝1:1.1)，加入溶剂2，室温条件下反应，结果如下：
N/A表示未检测
2)以乙酸乙酯作为溶剂1进行成盐反应
成盐步骤：取式(I)化合物约50mg,60℃下溶于2.0mL乙酸乙酯中，分别加入不同的对离子酸溶液(摩尔比为碱:酸＝1:1.1)，加入溶剂2，室温条件下反应，结果如下：
N/A表示未检测
3)以乙腈作为溶剂1进行成盐反应成盐步骤：取式(I)化合物约50mg，60℃下溶于2.0mL乙腈中，分别加入不同的对离子酸溶液(摩尔比为碱:酸＝1:1.1)，加入溶剂2，室温条件下反应，结果如下：

N/A表示未检测
4)以丙酮为溶剂1进行成盐反应
成盐步骤：取式(I)化合物约50mg,60℃下溶于2.0mL丙酮中，分别加入不同的对离子酸溶液(摩尔比为碱:酸＝1:1.1)，室温条件下反应，结果如下：
N/A表示未检测
通过上述盐的筛选研究发现，式(I)化合物半富马酸盐(式(II)化合物)在多个溶剂体系中均可制备得到，且多呈现结晶形式，结晶度良好。在相同条件体系下，其他酸多无法成盐、或成盐后为无定型状态或结晶度较差。
实施例2：结晶形式的式(I)化合物半富马酸盐的制备
取式(I)化合物约5.3g，加入220mL甲醇，60℃溶清；取约0.93g富马酸，加入10.6mL甲醇，60℃溶清；搅拌条件下将富马酸溶液滴加至式(I)化合物溶液中，室温搅拌1.5h，转移体系至4℃继续搅拌4.5h，抽滤，40℃真空干燥过夜，得到结晶形式的式(I)化合物半富马酸盐。
将上述结晶形式的式(I)化合物半富马酸盐暴露于相对湿度为75％RH、温度为30℃的环境中2天，制得半富马酸盐Form 1。
将上述结晶形式的式(I)化合物半富马酸盐暴露于相对湿度≤10％ RH、温度为30℃的环境中2天，制得半富马酸盐Form 2。
其中，半富马酸盐Form 1的XRPD数据如下表所示：

其XRPD图谱如图1所示。
其TGA图谱如图2所示。
其DSC图谱如图3所示。
其DVS图谱如图4所示。
其固态1H-NMR谱如图5所示，显示式(I)化合物和富马酸的摩尔比为1:0.5。
其中，半富马酸盐Form 2的XRPD数据如下表所示：

其XRPD图谱如图6所示。
其TGA图谱如图7所示。
其DSC图谱如图8所示。
其DVS图谱如图9所示。
其固态1H-NMR谱如图10所示，显示式(I)化合物和富马酸的摩尔比为1:0.5。
实施例3-5：结晶形式的式(I)化合物半富马酸盐的制备
取式(I)化合物约50mg，60℃下溶于2.0mL溶剂3中，取约35mg富马酸，60℃下溶于0.1mL有机溶剂中，将富马酸溶液滴加到式(I)化合物溶液中，室温条件下反应，选择性加入溶剂4，室温搅拌过夜，降温至4℃，继续搅拌3天，离心，真空干燥过夜，结果如下：
实施例6：结晶形式的式(I)化合物半富马酸盐的制备
取式(I)化合物半富马酸盐粗产物约30mg，46℃下加入四氢呋喃后溶清，将正庚烷滴加至溶清液中，置于室温下搅拌，析出固体后，离心，40℃真空干燥过夜，得到结晶形式的式(I)化合物半富马酸盐。
实施例7：结晶形式的式(I)化合物半富马酸盐的制备
将溶清液加入正庚烷中，其他同实施例6，得到结晶形式的式(I)化合物半富马酸盐。
实施例8-10：结晶形式的式(I)化合物半富马酸盐的制备
取式(I)化合物半富马酸盐粗产物约30mg，加入相应体积的溶剂后，升温至50℃溶清，置于4℃下搅拌，析出固体，离心，40℃真空干燥过夜，结果如下：
实施例11-34：结晶形式的式(I)化合物半富马酸盐的制备
取式(I)化合物半富马酸盐粗产物约50mg，加入相应体积的溶剂9后，形成悬浮液，将悬浮液浆液置于相应温度下搅拌5天，离心，40℃真空干燥过夜，结果如下：

实施例35：半富马酸盐Form 3的制备(乙醇溶剂合物)
取半富马酸盐Form 2约100mg，装于离心管中，于室温下置于装有乙醇的瓶中静置6天，经检测，得到半富马酸盐Form 3。
其XRPD数据如下表所示：

其XRPD图谱如图11所示。
其TGA图谱如图12所示，135℃之前失重约4.5％。
其DSC图谱如图13所示。
其固态1H-NMR谱如图14所示，显示式(I)化合物和富马酸的摩尔比为1:0.5。
实施例36：半富马酸盐Form 4的制备(水合物)
取式(I)化合物半富马酸盐粗产物约100mg，加入0.5mL仲丁醇和0.4mL乙酸乙酯，混悬液置于室温下晶浆2天，离心，所得固体在40℃下真空干燥过夜，经检测，得到半富马酸盐Form 4。
其XRPD数据如下表所示：

其XRPD图谱如图15所示。
其TGA图谱如图16所示，130℃之前失重约3.7％，约含1.5个水分子。
其DSC图谱如图17所示。
其DVS图谱如图18所示。
其固态1H-NMR谱如图19所示，显示式(I)化合物和富马酸的摩尔比为1:0.5。
实施例37-40：半富马酸盐Form 4的制备(水合物)
取半富马酸盐Form 2约50mg，加入相应体积的溶剂10后，形成悬浮液，将悬浮液浆液置于相应温度下搅拌5天，离心，40℃真空干燥过夜，结果如下：
实施例41：半富马酸盐Form 4的制备(水合物)
取式(I)化合物半富马酸盐粗产物约30mg，加入1.0mL丙酮，50℃加热溶清，置于室温下敞口挥发干，得到半富马酸盐Form 4。
实施例42：半富马酸盐Form 4的制备(水合物)
取式(I)化合物半富马酸盐粗产物约30mg，加入1.0mL乙酸乙酯，50℃加热溶清，置于40℃条件下敞口挥发干，得到半富马酸盐Form 4。
实施例43：半富马酸盐Form 4的制备(水合物)
取式(I)化合物半富马酸盐粗产物约30mg，加入0.5mL乙酸乙酯和0.1mL甲苯，50℃加热溶清，过滤，转至4℃下过夜，析出固体，离心，所得固体在40℃下真空干燥过夜，得到半富马酸盐Form 4。
实施例44：半富马酸盐Form 1单晶的制备
取半富马酸盐Form 1约30mg，放于玻璃小瓶中，加入0.5mL丁酮和0.5mL乙酸异丙酯，溶清，室温小孔挥发，得到块状晶体，为半富马酸盐Form 1的单晶，其单晶模拟图谱如图20所示。
其单晶结构数据如下表所示：

实施例45：式(I)化合物L-酒石酸盐的制备
取式(I)化合物约200mg溶于2.0mL溶剂12中；取L-酒石酸约45mg溶于0.8mL溶剂13中，室温下将L-酒石酸溶液滴加至搅拌中的式(I)化合物溶液中，转至4℃下搅拌过夜(约16小时)，析出固体得到混悬液，在混悬液中加入2.0mL溶剂14，置于4℃下搅拌过夜(约16小时)，离心，所得固体在室温下真空干燥过夜(约16小时)，结果如下：
实施例46：L-酒石酸盐Form 1的制备
取式(I)化合物约200mg溶于2.0mL甲苯中；取L-酒石酸约45mg溶于0.8mL乙醇中，室温下将L-酒石酸溶液滴加至搅拌中的式(I)化合物溶液中，转至4℃下搅拌过夜(约16小时)，析出固体得到混悬液，在混悬液中加入2.0mL异丙醚，置于4℃下搅拌过夜(约16小时)，离心，所得固体在室温下真空干燥过夜(约16小时)，得的L-酒石酸盐Form 1。
其XRPD数据如下表所示：
其XRPD图谱如图21所示。
其TGA图谱如图22所示。
其DSC图谱如图23所示。
其DVS图谱如图24所示。
其固态1H-NMR谱如图25所示，显示式(I)化合物和L-酒石酸的摩尔比为1:1。
实施例47-53：L-酒石酸盐Form 1的制备
取式(I)化合物L-酒石酸盐粗产物约20mg，悬浮在0.5mL溶剂18中，将悬浮液在适合温度下搅拌约3天后，分离固体，结果如下表：
实施例54：L-酒石酸盐Form 1的制备
取式(I)化合物L-酒石酸盐粗产物约20mg，溶解在0.1-3.0mL甲醇中,过滤后将滤液装入小瓶，用封口膜密封装有滤液的小瓶，放置在室温下针孔缓慢挥发，得到L-酒石酸盐Form 1。
实施例55：L-酒石酸盐Form 1的制备
取式(I)化合物L-酒石酸盐粗产物约20mg，悬浮于1.0mL乙醇中，然后将悬浮液加热至50℃，平衡约两小时，过滤，滤液在水浴中缓慢冷却至室温，分离固体，得到L-酒石酸盐Form 1。
实施例56：L-酒石酸盐Form 1的制备
取式(I)化合物L-酒石酸盐粗产物约20mg，悬浮在0.5mL体积比为1:9的氯仿和正庚烷混合溶剂中，在温度循环条件下(50℃保持120min，然后以0.1℃/min的降温速率将温度降至5℃，接着保持120min，重复2次)对悬浮液进行磁力搅拌，分离固体，得到L-酒石酸盐Form 1。
实施例57：L-酒石酸盐Form 2的制备
取L-酒石酸盐Form 1约20mg至3ml小瓶中，将其置于RT-97％相对湿度条件下7天，得到L-酒石酸盐Form 2。
其XRPD数据如下表所示：

其XRPD图谱如图26所示。
其TGA图谱如图27所示。
其DSC图谱如图28所示。
其固态1H-NMR谱如图29所示，显示式(I)化合物和L-酒石酸的摩尔比为1:1。
实施例58：L-酒石酸盐Form 3的制备
取L-酒石酸盐Form 1约20mg悬浮在0.5mL体积比为1:4的二甲基四氢呋喃和烷烃类溶剂的混合溶剂中，将悬浮液在4-8℃下搅拌约3天后，分离固体，得到L-酒石酸盐Form 3。
其XRPD数据如下表所示：

其XRPD图谱如图30所示。
其TGA图谱如图31所示。
其DSC图谱如图32所示。
其固态1H-NMR谱如图33所示，显示式(I)化合物和L-酒石酸的摩尔比为1:1。
实施例59：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约200mg溶解于1mL体积比为1:1的丙酮和甲基叔丁基醚混合溶剂中，过滤以获得澄清溶液，然后将澄清溶液放入用封口膜密封的20mL小瓶中，封口膜上有针孔，放置在室温下针孔缓慢挥发，1天后，析出固体，分离，得到L-酒石酸盐Form 4。
其XRPD数据如下表所示：

其XRPD图谱如图34所示。 其TGA图谱如图35所示。
其DSC图谱如图36所示。
其固态1H-NMR谱如图37所示，显示式(I)化合物和L-酒石酸的摩尔比为1:1。
实施例60-63：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约20mg，悬浮在0.5mL溶剂21中，并引入少量L-酒石酸盐Form 4作为晶种，将悬浮液在室温下磁力搅拌约3天后，分离固体，结果见下表：
实施例64：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约1.0g，悬浮在25mL体积比为1:6的丙酮/正庚烷混合溶剂中，并引入少量L-酒石酸盐Form 4作为晶种，悬浮液发生团聚现象，室温搅拌过夜(约16小时)，分离固体，得到L-酒石酸盐Form 4。
实施例65：L-酒石酸盐Form 4的制备
将室温搅拌替换为50℃搅拌，其他同实施例64，制备得到L-酒石酸盐Form4。
实施例66：L-酒石酸盐Form 4的制备
取式(I)化合物约1.5g和L-酒石酸约320.58mg悬浮在50mL体积比1:6的丙酮/正庚烷混合溶剂中，并引入少量L-酒石酸盐Form 4作为晶种，室温搅拌约2天，分离固体，得到L-酒石酸盐Form 4。
实施例67：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约20mg悬浮在0.5mL甲基叔丁基醚中，将悬浮液在4-8℃下搅拌约3天后，分离固体，得到L-酒石酸盐Form 4。
实施例68：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约20mg悬浮在0.5mL体积比为1:6的丁酮和正庚烷混合溶剂中，将悬浮液在室温下搅拌约3天后，分离固体，得到L-酒石酸盐Form 4。
实施例69-71：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约20mg溶解在0.1-0.8mL溶剂24中，过滤以获得澄清溶液，然后将该溶液放入含有3mL正己烷的20mL小瓶中，用瓶盖密封20ml小瓶，并保持在室温下，以便有足够的时间让有机蒸汽与溶液相互作用，结果如下表。

实施例72：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约20mg至3mL小瓶中，将其置于含有3mL丙酮或乙酸乙酯的20mL小瓶中，用盖子密封20mL小瓶，并在室温条件下保持7天，使溶剂蒸汽与样品相互作用，分离固体，得到L-酒石酸盐Form 4。
实施例73：L-酒石酸盐Form 4的制备
取L-酒石酸盐Form 1约20mg溶解0.1-3.0mL乙酸乙酯中，过滤后得滤液，在小瓶中加入约1～2mg乙基纤维素，用封口膜密封装有滤液的小瓶，并扎小孔，在室温条件下缓慢蒸发，分离固体，得到L-酒石酸盐Form 4。
实施例74：L-酒石酸盐Form 5的制备
取L-酒石酸盐Form 1约20mg悬浮在0.5mL体积比为1:1的异丙醇/甲基叔丁基醚混合溶剂中，将悬浮液在室温下搅拌约3天后，分离固体，得到L-酒石酸盐Form 5。
其XRPD数据如下表所示：

其XRPD图谱如图38所示。
实施例75：L-酒石酸盐Form 6的制备
取L-酒石酸盐Form 1约20mg，加热升温至70℃并持续10分钟，得到式L-酒石酸盐Form 6。
其XRPD数据如下表所示：
其XRPD图谱如图39所示。
实施例76：L-酒石酸盐无定型的制备
取L-酒石酸盐Form 4约300mg，加入30.0mL二氯甲烷，超声，过滤，将滤液在60℃下减压浓缩干，得到L-酒石酸盐无定型。
其XRPD图谱如图40所示。
其TGA图谱如图41所示。
其DSC图谱如图42所示。
其DVS图谱如图43所示。
其固态1H-NMR谱如图44所示，显示式(I)化合物和L-酒石酸的摩尔比为1:1。
实施例77：盐酸盐Form 1的制备
取式(I)化合物约100mg溶于1.0mL乙酸丁酯中，得到化合物溶液；取盐酸约14μL溶于0.2mL甲醇中，得到盐酸溶液；室温下将甲醇溶液滴加至搅拌中的乙酸丁酯溶液中，析出固体得到混悬液；将混悬液置于4℃下搅拌过夜，离心，所得固体在室温下真空干燥过夜得到盐酸盐Form 1。
其XRPD数据如下表所示：
其XRPD图谱如图45所示。
其TGA图谱如图46所示。
其DSC图谱如图47所示。
其DVS图谱如图48所示。
其高效液相(HPLC)检测结果显示式(I)化合物和盐酸的摩尔比为1:1。
实施例78：甲磺酸盐Form 1的制备
取式(I)化合物约50mg溶于0.5mL甲苯中，得到化合物溶液；取甲磺酸约7mg用0.1mL甲苯稀释，得到甲磺酸溶液；室温下将甲磺酸溶液滴加至搅拌中的化合物溶液中，转至4℃下搅拌过夜，析出固体得到混悬液；在混悬液中加入4.0mL异丙醚，继续在4℃下搅拌过夜(约16小时)，离心，所得固体在室温下真空干燥过夜，得到甲磺酸盐Form 1。
其XRPD数据如下表所示：

其XRPD图谱如图49所示。
实施例79：甲磺酸盐Form 2的制备
取式(I)化合物约200mg溶于2.0mL甲苯中，得到化合物溶液；取甲磺酸约29mg用0.4mL甲苯稀释，得到甲磺酸溶液；室温下将甲磺酸溶液滴加至搅拌中的化合物溶液中，转至4℃下搅拌过夜，析出固体得到混悬液；在混悬液中加入2.0mL异丙醚，继续在4℃下搅拌过夜(约16小时)，离心，所得固体在室温下真空干燥过夜，得到甲磺酸盐Form 2。
其XRPD数据如下表所示：

其XRPD图谱如图50所示。
其TGA图谱如图51所示。
其DSC图谱如图52所示。
其DVS图谱如图53所示。
其固态1H-NMR谱显示式(I)化合物和甲磺酸的摩尔比为1:1。
实施例80：甲磺酸盐Form 3的制备
取式(I)化合物约500mg溶于5.0mL甲苯中，得到化合物溶液；取甲磺酸约72mg用1.0mL甲苯稀释，得到甲磺酸溶液；室温下将甲磺酸溶液滴加至搅拌中的化合物溶液中，转至4℃下搅拌过夜，析出固体得到混悬液；在混悬液中加入5.0mL异丙醚，继续在4℃下搅拌过夜(约16小时)，离心，所得固体在室温下真空干燥过夜，得到甲磺酸盐Form 3。
其XRPD数据如下表所示：

其XRPD图谱如图54所示。
实施例81：草酸盐Form 1的制备
取式(I)化合物约50mg，溶于0.2mL丙酮中，得到化合物溶液；取草酸约6.7mg，溶于0.1mL丙酮中，得到草酸溶液；室温下，将草酸溶液滴加至化合物溶液中，置于室温下搅拌过夜(约16小时)，析出固体，得到混悬液；将混悬液中加入2.0mL异丙醚，4℃搅拌过夜，离心，所得固体在室温下真空干燥过夜，得到草酸盐Form 1。
其XRPD数据如下表所示：

其XRPD图谱如图55所示。
实施例82：草酸盐Form 2的制备
取式(I)化合物约200mg，溶于0.8mL丙酮中，得到化合物溶液；取草酸约38mg，溶于0.4mL丙酮中，得到草酸溶液；室温下，将草酸溶液滴加至化合物溶液中，置于室温下搅拌约0.5小时，析出固体，得到混悬液；将混悬液室温搅拌过夜，然后在混悬液中加入3.0mL异丙醚，4℃搅拌3天，离心，所得固体在室温下真空干燥过夜，得到草酸盐Form 2。
其XRPD数据如下表所示：

其XRPD图谱如图56所示。
其TGA图谱如图57所示。
其DSC图谱如图58所示。
其DVS图谱如图59所示。
其离子色谱(IC)显示式(I)化合物和草酸的摩尔比为1:1。
实验例
实验例1：稳定性研究
1.1半富马酸盐Form 1的稳定研究
取半富马酸盐Form 1适量，按照带包材和裸露两种情况，分别在高温(40℃和60℃)、高湿(25℃/90％RH)、加速(40℃/75％RH)条件下放置，30天后检测其XRPD，测试其晶型稳定性，结果如图61-62所示；并采用HPLC检测其含量，并计算有关物质的变化，稳定性结果如表1所示。
表1
结果表明：半富马酸盐Form 1在高温(40℃和60℃)、高湿(25℃/90％RH)和加速(40℃/75％RH)条件下放置30天，晶型基本保持不变，有关物质增加＜0.01％，具有良好的晶型稳定性和化学稳定性。
1.2半富马酸盐Form 2的稳定性研究
取半富马酸盐Form 2适量，在高温(敞口，60℃，<30％RH)条件下放置，10天后检测其XRPD，测试其晶型稳定性，结果如图62所示；并采用HPLC检测其含量，并计算有关物质的变化，稳定性结果如表2所示。
表2
结果表明，半富马酸盐Form 2在高温(敞口，60℃，<30％RH)条件下放置10天，晶型基本保持不变，有关物质增加＜0.05％，具有良好的晶型稳定性和化学稳定性。
1.3 L-酒石酸盐Form 1的稳定性研究
取L-酒石酸盐Form 1适量，分别在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置，1周后检测其XRPD，测试其晶型稳定性，结果如图63所示，并采用HPLC检测其含量，稳定性结果如表3所示。
表3
结果表明：L-酒石酸盐Form 1在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置1周，晶型基本保持不变，纯度基本保持不变，具有良好的晶型稳定性和化学稳定性。另外，将L-酒石酸盐Form 1在高温(60℃,<30％RH)条件下放置10天，其晶型基本保持不变(结果如图64所示)，有关物质增加＜0.05％，具有良好的晶型稳定性和化学稳定性。
1.4 L-酒石酸盐Form 4的稳定性研究
取L-酒石酸盐Form 4适量，分别在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置，1周后检测其XRPD，测试其晶型稳定性，结果如图65示，并采用HPLC检测其含量，稳定性结果如表4所示。
表4
结果表明：L-酒石酸盐Form 4在长期(25℃/60％RH)和加速(40℃/75％RH)条件下放置1周，晶型基本保持不变，纯度基本保持不变，具有良好的晶型稳定性和化学稳定性。另外，取L-酒石酸盐Form 4适量，按照带包材和裸露两种情况，分别在高温为40℃、50℃和55℃条件下放置30天，其晶型基本保持不变(结果如图66-68所示)，具有良好的晶型稳定性。
实验例2：粒度分布测试
分别称取适量待测样品半富马酸盐Form 1、半富马酸盐Form 2和草酸盐Form 2，加入10mL纯化水和两滴吐温80，摇匀后，超声10s，然后加入到马尔文激光粒度仪内(MS3000)检测。粒度分布测试结果如表5所示。半富马酸盐Form 1、半富马酸盐Form 2和草酸盐Form 2的PSD图分别如图69、图70和图71所示。
表5
结果表明，半富马酸盐Form 1和半富马酸盐Form 2的粒径分布较好，均接近正态分布。草酸盐Form 2的粒径偏大，分布较差。
实验例3：流动性测试
使用皓宇科技有限公司的HYL-1001型多功能粉体物理特性仪检测晶型的休止角。结果显示，草酸盐Form 2的休止角为58.10°，L-酒石酸盐Form 1和L-酒石酸盐Form 4的休止角分别为40.20°和32.73°，按照表6中粉体学指标的评价标准可知，L-酒石酸盐Form 4和L-酒石酸盐Form 1的流动性均优于草酸盐Form 2。
表6
实验例4：溶解度研究
分别称取半富马酸盐Form 1、半富马酸盐Form 2和草酸盐Form 2各约50mg，分别加入50mL pH4.5缓冲液(含0.5％吐温80)，置于37℃的摇床中振摇，24小时后采用HPLC法检测其平衡溶解度，24小时溶解度结果如表7。
表7
结果表明：相对于草酸盐Form 2，半富马酸盐Form 1和半富马酸盐Form 2在pH4.5缓冲溶液中，具有更高的溶解度。
实验例5：片剂的制备
按照表8的片剂处方，制备分别包含各盐型和其晶型的片剂。具体地，将各组分混合，涡旋混匀，使用红外压片机压(2Mpa的压力下保持2min压成片)制所有混合物以制备片剂。
表8
实验例6：片剂的可压性
分别取制备好的半富马酸盐Form 1片剂、半富马酸盐Form 2片剂和草酸盐Form 2片剂，用硬度仪检测直径和硬度，用游标卡尺检测厚度，并计算抗张强度。结果显示，半富马酸盐Form 1片剂的平均抗张强度为1.20MPa、半富马酸盐Form 2的平均抗张强度为1.34MPa，而草酸盐Form 2的平均抗张强度为1.14MPa。在一定的压力下，抗张强度越大，可压性越好。因此相对于草酸盐Form 2制备成的片剂，半富马酸盐Form 1制备成的片剂和半富马酸盐Form 2制备成的片剂的具有更有的可压性。
实验例7：片剂的溶出
分别取制备好的半富马酸盐Form 1片剂、半富马酸盐Form 2片剂和草酸盐Form 2片剂，测试其体外溶出度。
溶出介质：pH1.2溶液(含0.5％吐温80)
介质体积：900mL
溶出温度：37℃
溶出方法：浆法，75rpm
检测时间：10min、15min、45min、60min、120min、150min(150rpm)
溶出结果如下表9所示。
表9

结果表明，相对于草酸盐Form 2而言，以半富马酸盐Form 1和半富马酸盐Form 2作为活性成分的片剂，具有更好的体外溶出。
实验例8：不同形式的式(I)化合物在SD大鼠体内的药代动力学研究
8.1式(I)化合物游离态、式(I)化合物半富马酸盐和式(I)化合物L-酒石酸盐在SD大鼠体内的药代动力学研究
实验目的：通过动物PK研究，比较式(I)化合物游离态和半富马酸盐Form 1、半富马酸盐Form 2和L-酒石酸盐Form 4在SD体内的药代动力参数。
实验动物：动物种属及品系:大鼠，SD
性别及年龄:雄性，6～8周。
实验方法及数据处理：
取适量式(I)化合物游离态和半富马酸盐Form 1、半富马酸盐Form 2和L-酒石酸盐Form 4(均以游离碱的含量计)，将其在适量溶媒中溶解混合，其中注射给药溶媒(IV)为5％ DMSO+10％ Solutol+85％生理盐水，口服给药溶媒(PO)为1％吐温80+99％的0.5％ MC(4000cP)水溶液，按照表10给药方案进行给药，定时采集样本，并计算药代动力学参数。SD大鼠体内的药代动力学研究结果如表11所示。
表10
表11

F＝(AUC(0-t)(PO)*Dose(IV))/(AUC_(0-t)(IV)*Dose(PO))*100％)
结果表明，以2mg/kg的剂量，分别单次经口灌胃给予正常大鼠半富马酸盐Form 1和式(I)化合物游离态，半富马酸盐Form 1的血药浓度(135.19ng/mL)为式(I)化合物游离态血药浓度(71.70ng/mL)的1.9倍；半富马酸盐Form 1的曲线面积(2276.92h*ng/mL)为式(I)化合物游离态曲线面积(1236.31h*ng/mL)的1.8倍，半富马酸盐Form 1的口服绝对生物利用度(36.01％)为式(I)化合物游离态(11.21％)的3.2倍。
同样条件下，半富马酸盐Form 2的口服绝对生物利用度(30.40％)为式(I)化合物游离态(11.21％)的2.7倍。
同样条件下，L-酒石酸盐Form 4的口服绝对生物利用度(42.58％)为式(I)化合物游离态(11.21％)的3.8倍。
综上，以2mg/kg的剂量口服给药时，半富马酸盐Form 1、半富马酸盐Form 2和L-酒石酸盐Form 4的血药浓度和曲线面积分别相对于式(I)化合物游离态具有明显的优势，且半富马酸盐Form 1、半富马酸盐Form 2和L-酒石酸盐Form 4的口服绝对生物利用度相对于式(I)化合物游离态具有明显的优势。
8.2式(I)化合物游离态和L-酒石酸盐Form 1在SD大鼠体内的药代动力学研究
实验目的：通过动物PK研究，比较式(I)化合物游离态和L-酒石酸盐Form 1在SD体内的药代动力参数。
实验动物：动物种属及品系:大鼠，SD
性别及年龄:雄性，6～8周。
体重：200～250g。
实验方法及数据处理：取适量式(I)化合物游离态和L-酒石酸盐Form 1(以游离碱的含量计)，将其在适量溶媒中溶解混合，其中注射给药溶媒(IV)为5％DMSO+10％ Solutol+85％生理盐水，口服给药溶媒(PO)为1％吐温80+99％的0.5％ MC(4000cP)水溶液，按照表12的给药方案进行给药，定时采集样本，并计算药代动力学参数。SD大鼠体内的药代动力学研究结果如表13所示。
表12
表13
F＝(AUC(0-t)(PO)*Dose(IV))/(AUC_(0-t)(IV)*Dose(PO))*100％)
结果表明，以10mg/kg的剂量，分别单次经口灌胃给予正常大鼠L-酒石酸盐Form 1和式(I)化合物游离态，L-酒石酸盐Form 1的血药浓度(1114.38ng/mL)为式(I)化合物游离态血药浓度(748.41ng/mL)的1.5倍，L-酒石酸盐Form 1的曲线面积(34189.97h*ng/mL)为式(I)化合物游离态曲线面积(18972.79h*ng/mL)的1.8倍，L-酒石酸盐Form 1的口服绝对生物利用度(43.17％)为式(I)化合物游离态(16.78％)的2.6倍。
综上，以10mg/kg的剂量口服给药时，L-酒石酸盐Form 1的血药浓度和曲线面积相对于式(I)化合物游离态具有明显的优势，且L-酒石酸盐Form 1的口服绝对生物利用度相对于与式(I)化合物游离态具有明显的优势。
实验例9：半富马酸盐Form 1和L-酒石酸盐Form 4对人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤生长抑制作用的研究
实验目的：使用人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤模型，对半富马酸盐Form 1和L-酒石酸盐Form 4的抗肿瘤活性进行评价。
实验动物：动物种属及品系:Balb/c裸鼠
性别及年龄:雌性，7-9周龄。
受试药物的配制：溶媒：1％吐温80+99％MC溶液(0.5％)。
溶媒的配制(0.5％ MC溶液)及保存：移取适量的MC到合适容器中，加入计算量的ddH2O，搅拌、涡旋或超声使充分溶解，密封备用。
给药溶液的配制及保存：称取一定量半富马酸盐Form 1(以游离碱的含量计)至合适的容器中，加入计算量溶媒，搅拌、涡旋或超声混匀，得所需浓度给药溶液，记录物理性状，4℃，现配现用。
实验方法及数据处理：
1)模型建立
MDA-MB-231细胞培养于L-15培养基，含10％胎牛血清FBS(GIBCO，美国)，细胞在无二氧化碳的闭口培养瓶并放置于培养箱37℃培养。
收集对数生长期MDA-MB-231细胞，重悬于无血清L-15基础培养基中，1:1加入Matrigel中，调整细胞悬液浓度至6.67×107/ml。在无菌条件下，接种0.15mL细胞悬液至小鼠右侧背部皮下，接种浓度为1×107/0.15mL/mouse。
2)分组及给药观察
在肿瘤平均体积达到119mm3左右时，将动物按肿瘤体积随机分组，使各组肿瘤体积差异小于均值的10％，分组当日记为Day0，并按照动物体重或每只动物固定给药体积开始给药，给药方案如表14。
表14
给药期间，个别动物体重与Day0相比下降超过15％(BWL≥15％)，将做停药处理，直至动物体重恢复后(BWL<15％)，恢复给药。
实验周期为4周，实验期间每周2次测定动物体重和肿瘤体积，每日1次观察记录动物临床症状。
3)评价指标和数据分析
肿瘤体积(tumor volume，TV)计算公式为：1/2×a×b2，其中a、b分别为肿瘤测量的长和宽。
相对肿瘤体积(relative tumor volume，RTV)计算公式为：Vt/V0，其中V0为分组时的肿瘤体积，Vt为每一次测量时的肿瘤体积。
相对肿瘤增值率T/C(％)计算公式为：TRTV/CRTV×100％，其中TRTV为治疗组RTV，CRTV为阴性对照组RTV。
肿瘤体积抑瘤率(TGI)计算公式为：(TVC-TVT)/TVC×100％，其中TVC为空白溶剂对照组平均肿瘤体积，TVT为治疗组平均肿瘤体积
抑瘤率(％)计算公式为：(TWC-TWT)/TWC×100％，其中TWC为阴性对照组平均肿瘤重量，TWT为治疗组平均肿瘤重量。
动物体重变化BWC(％)计算公式为：(BWt-BW0)/BW0×100％，其中BWt为每次测量时动物体重，BW0为分组时动物体重。
根据中国NMPA《细胞毒类抗肿瘤药物非临床研究技术指导原则》(2006年11月)，T/C(％)≤40％并经统计学分析P<0.05为有效。
本研究中，实验数据均以Mean±SEM表示。
以时间点为X轴，肿瘤体积(mm3)为Y轴绘制肿瘤生长曲线；以时间点为X轴，动物体重(g)为Y轴绘制动物体重变化曲线。组间比较采用双尾t-检验，P<0.05为显著性差异，P<0.01为极显著差异。
对人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤生长抑制作用的研究结果如表15所示。
表15
*p<0.05，**p<0.01，***p<0.001
结果表明，Day 28实验结束时，溶媒对照组平均肿瘤体积为564.69±26.44mm3，半富马酸盐Form 1(10mg/kg)组的平均肿瘤体积为138.60±14.36mm3，肿瘤体积抑瘤率TGI(％)为75.46％，给药后的相对肿瘤增殖率T/C为23.91％，小于40％,能显著抑制人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤的生长。L-酒石酸盐Form 4(10mg/kg)组的平均肿瘤体积为116.17±7.25，肿瘤体积抑瘤率TGI(％)为79.43％，给药后的相对肿瘤增殖率T/C为20.37％，小于40％,能显著抑制人乳腺癌MDA-MB-231细胞Balb/c裸鼠异种移植瘤的生长。
实验例10：半富马酸盐Form 1对人急性髓系白血病MOLM16细胞CB-17SCID小鼠异种移植瘤生长抑制作用的研究
实验目的：使用人急性髓系白血病MOLM16细胞CB-17SCID小鼠异种移植瘤模型，半富马酸盐Form 1的体内抗肿瘤活性进行评价。
实验动物：动物种属及品系:CB17-SCID
性别及年龄:雌性，6-7周龄。
受试药物的配制：溶媒：1％吐温80+99％MC溶液(0.5％)。
溶媒的配制(0.5％ MC溶液)及保存：移取适量的MC到合适容器中，加入计算量的ddH2O，搅拌、涡旋或超声使充分溶解，密封备用。
给药溶液的配制及保存：称取一定量对半富马酸盐Form 1(以游离碱的含量计)至合适的容器中，加入计算量溶媒，搅拌、涡旋或超声混匀，得所需浓度给药溶液，记录物理性状，室温，现配现用。
实验方法及数据处理：
1)模型建立
MOLM16细胞培养于含20％ FBS的RPMI-1640培养基中，维持在5％ CO2的37℃饱和湿度培养箱中。
收集对数生长期MOLM16细胞，重悬于RPMI-1640基础培养基中，1:1加入基质胶，调整细胞浓度至1×107/mL。在无菌条件下，接种0.1mL细胞悬液至裸鼠右侧背部皮下，接种浓度为1×106/0.1mL/mouse。
2)分组及给药观察
在肿瘤平均体积达到106mm3左右时，将动物按肿瘤体积随机分组，使各组肿瘤体积差异小于均值的10％，分组当日记为Day0，并按照动物体重或每只动物固定给药体积开始给药。给药方案如表16所示。
表16
给药期间，个别动物体重与Day0相比下降超过15％(BWL≥15％)，将做停药处理，直至动物体重恢复后(BWL<15％)，恢复给药。
实验周期为2周，实验期间每周2次测定动物体重和肿瘤体积，每日1次观察记录动物临床症状。
3)评价指标和数据分析
评价指标和数据分析方法同实验例9一致。
对人急性髓系白血病MOLM16细胞CB-17SCID小鼠异种移植瘤生长抑制作用的研究结果如表17所示。
表17

*p<0.05，**p<0.01，***p<0.001
结果表明，Day 14实验结束时，溶媒对照组平均肿瘤体积为1788.88±132.86mm3，半富马酸盐Form 1(1mg/kg、3mg/kg和10mg/kg)组的平均肿瘤体积分别为172.85±14.02mm3、120.33±3.15mm3和64.08±7.11mm3，相对肿瘤增值率T/C分别为9.61％(TGI＝90.34％，P<0.001)、6.77％(TGI＝93.27％，P<0.001)和3.62％(TGI＝96.42％，P<0.001)，小于40％,均能显著抑制人白血病MOLM16细胞CB-17SCID小鼠异种移植瘤的生长。
实验例11：半富马酸盐Form 1对人间变大细胞淋巴瘤SU-DHL-1细胞CB-17SCID小鼠异种移植瘤生长抑制作用的研究
实验目的：使用人间变大细胞淋巴瘤SU-DHL-1细胞CB-17SCID小鼠异种移植瘤模型，对半富马酸盐Form 1的抗肿瘤活性进行评价。
实验动物：动物种属及品系:CB-17SCID小鼠。
性别及年龄:雌性，6-8周龄。
体重：18-22g，偏差为体重均值的±20％。
受试药物的配制：溶媒：1％吐温80+99％MC溶液(0.5％)。
溶媒的配制(0.5％ MC溶液)及保存：移取适量的MC到合适容器中，加入计算量的ddH2O，搅拌、涡旋或超声使充分溶解，密封备用。
给药溶液的配制及保存：称取一定量半富马酸盐Form 1(以游离碱的含量计)至合适的容器中，加入计算量溶媒，搅拌、涡旋或超声混匀，得所需浓度给药溶液，记录物理性状，4℃，现配现用。
实验方法及数据处理：
1)模型建立
SU-DHL-1细胞培养于含10％ FBS的RPMI-1640培养基中，维持在5％ CO2的37℃饱和湿度培养箱中。
收集对数生长期SU-DHL-1细胞，重悬于RPMI-1640基础培养基中，1:1加入基质胶，调整细胞浓度至3×107/mL。在无菌条件下，接种0.1mL细胞悬液至裸鼠右侧背部皮下，接种浓度为3×106/0.1mL/mouse。
2)分组及给药观察
在肿瘤平均体积达到106mm3左右时，将动物按肿瘤体积随机分组，使各组肿瘤体积差异小于均值的10％，分组当日记为Day0，并按照动物体重或每只动物固定给药体积开始给药。给药方案如表18所示。
表18
给药期间，个别动物体重与Day0相比下降超过15％(BWL≥15％)，将做停药处理，直至动物体重恢复后(BWL<15％)，恢复给药。
实验周期为3周，实验期间每周2次测定动物体重和肿瘤体积，每日1次观察记录动物临床症状。
3)评价指标和数据分析
评价指标和数据分析方法同实验例9一致。
对人间变大细胞淋巴瘤SU-DHL-1细胞CB-17 SCID小鼠异种移植瘤生长抑制作用的研究结果如表19所示。
表19
*p<0.05，**p<0.01，***p<0.001
结果表明，Day 21实验结束时，溶媒对照组平均肿瘤体积为2029.90±204.72mm3，半富马酸盐Form 1(10mg/kg)组的平均肿瘤体积为316.83±97.40mm3，Day 21时的相对肿瘤增值率T/C为15.53％(TGI＝84.39％，P<0.001),小于40％，半富马酸盐Form 1能显著抑制人间变大细胞淋巴瘤SU-DHL-1细胞CB-17 SCID小鼠异种移植瘤的生长。
实验例12：不同形式的式(I)化合物对人Molm16细胞在免疫缺陷鼠皮下移植瘤的药效学研究
实验目的：比较式(I)化合物游离态和L-酒石酸盐Form 1在NOD-SCID免疫缺陷鼠Molm16皮下肿瘤模型中的药效。
实验动物：动物种属及品系:小鼠，NOD-SCID
性别及年龄:雄性，6-8周。
受试化合物配制：溶媒的配制：取适量的Tween80加入99ml的0.5％MC，涡旋混匀，备用；
给药溶液的配制：称取一定量式(I)化合物游离态、L-酒石酸盐Form 1(以游离碱的含量计)至合适的容器中，加入计算量溶媒，搅拌混匀，得所需浓度给药溶液，现配现用。
实验方法及数据处理：
1)Molm16皮下异位肿瘤模型建立
Molm16细胞培养于含20％灭活胎牛血清(Gbico-10099141)、1％青/链霉素的1640培养基中，使用细胞培养瓶在含5％ CO2的37℃的细胞培养箱中连续培养；待细胞长至对数生长期时，收集细胞，并用PBS洗涤细胞一次，细胞计数，离心，最后用PBS重悬，加入50％ Matrigel(Cultrex PathClear BM,3432-010-01P)，调整细胞浓度后用于后续接种；每只小鼠右侧腋下接种3×106个细胞，接种体积为100ul。
2)动物分组及给药
细胞接种后第5天，测量肿瘤的长和宽，通过肿瘤体积计算公式：＝0.5a×b2，a和b分别表示肿瘤的长和宽，来计算肿瘤体积，并依据肿瘤体积进行分组。选取体积80mm3到120mm3的肿瘤随机分组，肿瘤体积每组均值为93mm3，然后进行给药，给药方案如表20所示。
表20
3)观察
每天监测动物的健康状况及死亡情况，例行检查包括观察受试物和药物对动物日常行为表现的影响如行为活动，摄食摄水量，体重变化(每周测量两次体重)，外观体征或其它不正常情况。
每周两次测量肿瘤直径，并计算肿瘤体积。
肿瘤体积的计算公式为：V＝0.5a×b2，a和b分别表示肿瘤的长和宽。
相对肿瘤增殖率T/C％＝TRTV/CRTV×100％(RTV＝Vt/V0)
抑瘤率TGI％＝(1-T/C)×100％
4)统计分析
采用Two-way ANOVA来比较体重和肿瘤体积。采用One-way ANOVA来比较实验终点时的肿瘤重量。所有数据将使用GraphPad Prism 8进行分析，p<0.05表示有统计学差异。
受试化合物在NOD-SCID免疫缺陷鼠Molm16皮下肿瘤模型中的药效如表21所示。
表21
注释:a.荷瘤第5天的肿瘤体积
b.荷瘤第21天的肿瘤体积
c.所有组均与溶媒组比，*P<0.05，**P<0.01，***P<0.001，****P<0.0001。
结果表明，口服连续给药21天后，所有给药组与溶媒相比，均产生了极显著性的抑瘤效果(p<0.0001)，TGI值分别为82.28％、95.41％、96.96％、91.79％、96.63％和98.35％；同等剂量下，L-酒石酸盐Form 1抑瘤效果优于式(I)化合物游离态。
实验例13：L-酒石酸盐Form 4对人胰腺癌BxPC-3细胞Balb/c裸鼠异种移植瘤生长抑制作用的研究实验目的：使用人胰腺癌BxPC-3细胞Balb/c裸鼠异种移植瘤模型，对L-酒石酸盐Form 4的抗肿瘤活性进行评价。
实验动物：动物种属及品系:Balb/c裸鼠
性别及年龄:雌性，9-11周龄。
体重：21-24g，偏差为体重均值的±20％。
受试药物的配制：溶媒：1％吐温80+99％MC溶液(0.5％)。
溶液配制及保存：溶媒的配制(0.5％ MC溶液)：移取适量的MC到合适容器中，加入计算量的ddH2O，搅拌、涡旋或超声使充分溶解，密封备用。
给药溶液的配制及保存：称取一定量L-酒石酸盐Form 4(以游离碱的含量计)至合适的容器中，加入计算量溶媒，搅拌、涡旋或超声混匀，得所需浓度给药溶液，记录物理性状。4℃，现配现用。
实验方法及数据处理：
1)模型建立
BxPC-3细胞培养于含10％ FBS的RPMI-1640培养基中，维持在5％ CO2的37℃饱和湿度培养箱中。
收集对数生长期BxPC-3细胞，重悬于RPMI-1640基础培养基中，1:1加入基质胶，调整细胞浓度至5×107/mL。在无菌条件下，接种0.1mL细胞悬液至裸鼠右侧背部皮下，接种浓度为5×106/0.1mL/mouse。
2)分组及给药观察
在肿瘤平均体积达到110mm3左右时，将动物按肿瘤体积随机分组，使各组肿瘤体积差异小于均值的10％，分组当日记为Day0，并按照动物体重或每只动物固定给药体积开始给药。给药方案如表22所示。
表22
给药期间，个别动物体重与Day0相比下降超过15％(BWL≥15％)，将做停药处理，直至动物体重恢复后(BWL<15％)，恢复给药。
实验周期为4周，实验期间每周2次测定动物体重和肿瘤体积，每日1次观察记录动物临床症状。
3)评价指标和数据分析
评价指标和数据分析方法同实验例9一致。
对人胰腺癌BxPC-3细胞Balb/c裸鼠异种移植瘤生长抑制作用的研究结果如表23所示。
表23
*p<0.05，**p<0.01，***p<0.001
结果表明，Day 28实验结束时，溶媒对照组平均肿瘤体积为1175.00±147.32mm3，L-酒石酸盐Form 4的3mg/kg组和10mg/kg组的平均肿瘤体积分别为379.47±65.56mm3和307.27±49.38mm3，肿瘤体积抑瘤率TGI(％)分别为67.70％和73.85％，给药后的相对肿瘤增殖率T/C分别为32.44％和26.78％，小于40％，L-酒石酸盐Form 4能显著抑制人胰腺癌BxPC-3细胞Balb/c裸鼠异种移植瘤的生长。
实验例14：L-酒石酸盐Form 4对人非小细胞肺癌NCI-H460细胞Balb/c裸鼠异种移植瘤生长抑制作用的研究
实验目的：使用人非小细胞肺癌NCI-H460细胞Balb/c裸鼠异种移植瘤模型，对L-酒石酸盐Form 4的抗肿瘤活性进行评价。
实验动物：动物种属及品系:Balb/c裸鼠
性别及周龄：雌性，8-9周龄。
体重：18-22g，偏差为体重均值的±20％。
受试药物的配制：溶媒：1％吐温80+99％MC溶液(0.5％)。
溶液配制及保存：溶媒的配制(0.5％ MC溶液)：移取适量的MC到合适容器中，加入计算量的ddH2O，搅拌、涡旋或超声使充分溶解，密封备用。
给药溶液的配制及保存：称取一定量L-酒石酸盐Form 4(以游离碱的含量计)至合适的容器中，加入计算量溶媒，搅拌、涡旋或超声混匀，得所需浓度给药溶液，记录物理性状。4℃，现配现用。
实验方法及数据处理：
1)模型建立
NCI-H460细胞培养于含10％ FBS的RPMI-1640培养基中，维持在5％ CO2的37℃饱和湿度培养箱中。
收集对数生长期NCI-H460细胞，重悬于RPMI-1640基础培养基中，1:1加入基质胶，调整细胞浓度至2×107/mL。在无菌条件下，接种0.1mL细胞悬液至裸鼠右侧背部皮下，接种浓度为2×106/0.1mL/mouse。
2)分组及给药观察
在肿瘤平均体积达到109mm3左右时，将动物按肿瘤体积随机分组，使各组肿瘤体积差异小于均值的10％，分组当日记为Day0，并按照动物体重或每只动物固定给药体积开始给药。给药方案如表24所示。
表24
给药期间，个别动物体重与Day0相比下降超过15％(BWL≥15％)，将做停药处理，直至动物体重恢复后(BWL<15％)，恢复给药。
实验周期为17天，实验期间每周2次测定动物体重和肿瘤体积，每日1次观察记录动物临床症状。
3)评价指标和数据分析
评价指标和数据分析方法同实验例9一致。
对人非小细胞肺癌NCI-H460细胞Balb/c裸鼠异种移植瘤生长抑制作用的研究结果如表25所示。
表25
p<0.05，**p<0.01，***p<0.001
结果表明，Day 17实验结束时，溶媒对照组平均肿瘤体积为2109.69±113.70mm3，L-酒石酸盐Form 4的3mg/kg组和10mg/kg组的平均肿瘤体积分别为803.58±42.42和710.49±52.36，肿瘤体积抑瘤率TGI(％)分别为61.91％和66.32％，给药后的相对肿瘤增值率TGI(％)分别为38.29％和33.68％，小于40％，L-酒石酸盐Form 4能显著抑制人非小细胞肺癌NCI-H460细胞Balb/c裸鼠异种移植瘤的生长。
上述实施例为本申请较佳的实施方式，但本申请的实施方式并不受上述实施例的限制，其他的任何未背离本申请的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本申请的保护范围之内。

Claims (36)

  1. 一种式(I)化合物的盐，
    所述盐为酸式盐，其中，所述式(I)化合物和酸的摩尔比为0.5-2。
  2. 根据权利要求1所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐选自富马酸盐、L-酒石酸盐、盐酸盐、甲磺酸盐和草酸盐。
  3. 根据权利要求1或2所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐选自富马酸盐。
    优选地，所述式(I)化合物和富马酸的摩尔比为1:0.5-1；更优选为1:0.5。
  4. 根据权利要求1-3中任一项所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐为半富马酸盐，结构式如式(II)所示：
    优选地，所述式(I)化合物半富马酸盐基本上以结晶形式或无定型形式存在。
  5. 根据权利要求4所述的式(I)化合物的盐，其特征在于，所述式(I)化合物半富马酸盐为结晶形式，使用Cu-Kα辐射，所述半富马酸盐的结晶形式以2θ角度表示的XRPD图谱在15.7°±0.2°、16.1°±0.2°和9.4°±0.2°中的至少一处具有特征峰。优选为二处或三处有特征峰。
    优选地，所述半富马酸盐的结晶形式以2θ角度表示的XRPD图谱在15.7°±0.2°、16.1°±0.2°和9.4°±0.2°处有特征峰。
    优选地，所述半富马酸盐的结晶形式的XRPD图谱还在14.0°±0.2°、17.1°±0.2°、17.9°±0.2°和19.2°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述半富马酸盐的结晶形式的XRPD图谱还在18.7°±0.2°、21.8°±0.2°、23.4°±0.2和27.1°±0.2°2θ中的至少一处具有特征峰。
  6. 根据权利要求5所述的式(I)化合物的盐，其特征在于，所述半富马酸盐的结晶形式为式(I)化合物半富马酸盐Form 1。
    优选地，所述半富马酸盐Form 1的XRPD图谱还在7.0°±0.2°、10.2°±0.2°、24.5°±0.2°和20.1°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述半富马酸盐Form 1具有基本上与图1相同的XRPD图谱。
    优选地，所述半富马酸盐Form 1为无水物。
    优选地，所述半富马酸盐Form 1的单晶晶体学参数为：单斜晶系，空间群为P21/n；晶胞参数为：α＝γ＝90°，β＝104.031(2)°。
    优选地，所述半富马酸盐Form 1具有基本如图20所示的XRD单晶模拟图谱。
  7. 根据权利要求5所述的式(I)化合物的盐，其特征在于，所述半富马酸盐的结晶形式为式(I)化合物半富马酸盐Form 2。
    优选地，所述半富马酸盐Form 2的XRPD图谱还在16.5°±0.2°、17.3°±0.2°、19.5°±0.2°、24.8°±0.2°和27.6°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述半富马酸盐Form 2具有基本上与图6相同的XRPD图谱。
    优选地，所述半富马酸盐Form 2为无水物。
  8. 根据权利要求5-7中任一项所述的结晶形式的式(I)化合物半富马酸盐的制备方法，所述制备方法包含以下方法中的任一种：
    1)步骤A:将适量式(I)化合物在溶剂3中溶解；
    步骤B:将适量富马酸在有机溶剂中溶解；
    步骤C:将上述两种溶液合并，搅拌析出或加入溶剂4后搅拌析出，形成混悬液；
    步骤D:继续搅拌上述混悬液，并将其在低温环境下搅拌析晶，分离、干燥，得到结晶形式的式(I)化合物半富马酸盐。
    优选地，所述溶剂3选自甲醇、乙酸乙酯、乙腈中的任一种或其混合。
    优选地，所述有机溶剂选自醇类。
    优选地，所述溶剂4选自水、异丙醚、异丙醇、正庚烷和甲基环己烷中的一种或其组合。
    优选地，所述式(I)化合物和富马酸的投料摩尔比为1：0.5-1.5。
    优选地，所述式(I)化合物和溶剂3的质量体积比(mg/ml)≤100:1；优选≤80:1；更优选≤50:1。
    2)将式(I)化合物半富马酸盐的粗产物在溶剂5中加热溶清，然后和溶剂6混合后，搅拌析晶，分离，干燥，得所述结晶形式的式(I)化合物半富马酸盐。
    优选地，所述溶剂5选自醇类、酮类、酯类、醚类、二甲亚砜和四氢呋喃中一种或其混合。
    优选地，所述溶剂6选自水、正庚烷、异丙醚、二氯甲烷、氯仿、甲苯、甲基环己烷中的一种或其混合。
    优选地，所述溶剂6和溶剂5的体积比≥1:1；更优选为≥1.5:1。
    优选地，所述加热溶清的温度≥40℃；更优选为50℃-60℃。
    3)将式(I)化合物半富马酸盐在溶剂7或溶剂7和溶剂8的混合溶剂体系中加热溶清，降温搅拌，析晶，分离，干燥，得所述结晶形式的式(I)化合物半富马酸盐。
    优选地，所述溶剂7选自醇类、酮类、酯类、醚类、二甲亚砜和四氢呋喃中一种或其混合；更优选地，所述溶剂7选自醇类、酮类；最优选地，所述醇类选自乙醇和甲醇中的一种或其组合；所述酮类选自丙酮。
    优选地，所述溶剂8选自水、正庚烷、异丙醚、二氯甲烷、氯仿、甲苯、甲基环己烷中的一种或其混合；更优选水和正庚烷中的一种或其混合。
    优选地，所述加热溶清的温度≥40℃；更优选为50℃-60℃。
    优选地，所述降温搅拌的温度≤30℃；更优选为0℃-20℃。
    4)在式(I)化合物半富马酸盐的粗产物中加入溶剂9形成混悬液，搅拌析晶，分离，干燥，得所述结晶形式的结晶形式的式(I)化合物半富马酸盐。
    优选地，所述溶剂9选自醇类、烷烃类、酯类、腈类、醚类、苯环类、酮类和水中的一种或其组合；更优选地，所述醇类选自甲醇、乙醇和异丙醇中的一种或其组合；所述烷烃类选自二氯甲烷、正庚烷、甲基环己烷中的一种或其组合；所述酯类选自乙酸乙酯、乙酸异丙酯中一种或其组合；所述腈类选自乙腈；所述醚类选自乙醚、异丙醚中的一种或其组合；所述苯环类选自甲苯；所述酮类选自丙酮和丁酮中的一种或其组合。
    优选地，所述溶剂9选自单一溶剂体系，比如甲醇、水、异丙醚、二氯甲烷、异丙醇、甲基叔丁基醚、甲苯、乙腈、乙醚中的一种。
    优选地，所述溶剂9为特定体积比的混合溶剂体系，比如甲醇和水的混合溶剂体系，其体积比为2：1；乙醇和水的混合溶剂体系，其体积比为3：1；丙酮和水的混合溶剂体系，其体积比为1：2；乙腈和水的混合溶剂体系，其体积比为1：4；乙酸乙酯的饱和水混合溶剂体系、乙醇和异丙醚的混合溶剂体系，其体积比为1：1；异丙醇和正庚烷的混合溶剂体系，其体积比为1：1；丁酮和甲基环己烷的混合溶剂体系，其体积比为1：5；异丙醚和乙酸乙酯的混合溶剂体系，其体积比为5：1、乙酸乙酯和二氯甲烷的混合溶剂体系，其体积比为1：5；乙酸异丙酯和甲基环己烷的混合溶剂体系，其体积比为1：5、乙醇和丙酮的混合溶剂体系，其体积比为4：1；异丙醚和乙腈的混合溶剂体系，其体积比为5：2、乙醚和甲苯的混合溶剂体系，其体积比为2：1、乙腈和甲苯的混合溶剂体系，其体积比为2：1、乙腈和乙酸乙酯的混合溶剂体系，其体积比为1：2；二氯甲烷和甲苯的混合溶剂体系，其体积比为1：10。
    优选地，所述式(I)化合物半富马酸盐的粗产物和溶剂9的质量体积比(mg/ml)≤150:1；更优选为≤120:1。
  9. 根据权利要求6所述的半富马酸盐Form 1的制备方法，其包括将权利要求8中任意方法得到结晶形式的式(I)化合物半富马酸盐暴露于相对湿度≥75％的环境中足够的时间，得到式半富马酸盐Form 1。
    优选地，所述暴露温度≤40℃。
    优选地，所述足够的时间是指XRPD检测后确定得到半富马酸盐Form 1为止；在一些实施例中，足够的时间一般为2天以上。
  10. 根据权利要求7所述的半富马酸盐Form 2的制备方法还在于提供一种半富马酸盐Form 2的制备方法，其包括将权利要求8中任意方法得到结晶形式的式(I)化合物半富马酸盐暴露于相对湿度≤10％的环境中足够的时间，得到半富马酸盐Form 2。
    优选地，所述暴露温度≥30℃。
    优选地，所述足够的时间是指XRPD检测后确定得到半富马酸盐Form 2为止；在一些实施例中，足够的时间一般为2天以上。
  11. 根据权利要求4所述的式(I)化合物的盐，其特征在于，所述式(I)化合物半富马酸盐为半富马酸盐乙醇溶剂合物Form 3，其特征在于，使用Cu-Kα辐射，所述半富马酸盐Form 3以2θ角度表示的XRPD图谱在6.8°±0.2°、9.4°±0.2°、15.7°±0.2°和15.9°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处具有特征峰。
    优选地，所述半富马酸盐Form 3的XRPD图谱还在17.8°±0.2°、18.5°±0.2°、19.0°±0.2°和22.1°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述半富马酸盐Form 3的XRPD图谱还在23.9°±0.2°、25.1°±0.2°、26.2°±0.2°和28.6°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述半富马酸盐Form 3具有基本上与图11相同的XRPD图谱。
    优选地，所述半富马酸盐Form 3中乙醇与式(I)化合物半富马酸盐的摩尔比为1:1。
  12. 根据权利要求4所述的式(I)化合物的盐，其特征在于，所述式(I)化合物半富马酸盐为半富马酸盐的水合物Form 4，其特征在于，使用Cu-Kα辐射，所述半富马酸盐Form 4以2θ角度表示的XRPD图谱在9.3°±0.2°、15.6°±0.2°、16.0°±0.2°、和21.9°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处有特征峰。
    优选地，所述半富马酸盐Form 4的XRPD图谱还在17.1°±0.2°、22.8°±0.2°、24.1°±0.2°和25.0°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述半富马酸盐Form 4的XRPD图谱还10.0°±0.2°、9.5°±0.2°、19.0°±0.2°和20.3°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述半富马酸盐Form 4具有基本上与图15相同的XRPD图谱。
  13. 根据权利要求1或2所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐选自L-酒石酸盐。
    优选地，所述式(I)化合物和L-酒石酸的摩尔比为1:0.5-2；更优选为1:1。
    优选地，所述式(I)化合物的盐选自L-酒石酸盐，结构式如式(III)所示:
    优选地，所述式(I)化合物L-酒石酸盐基本上以结晶形式或无定型形式存在。
  14. 根据权利要求13所述的式(I)化合物的盐，其特征在于，所述式(I)化合物L-酒石酸盐为L-酒石酸盐的Form 1，其特征在于，使用Cu-Kα辐射，所述L-酒石酸盐Form 1以2θ角度表示的XRPD图谱在7.8°±0.2°、14.6°±0.2°、18.3°±0.2°和23.6°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处有特征峰。
    优选地，所述L-酒石酸盐Form 1的XRPD图谱还在20.0°±0.2°、14.1°±0.2°、13.5°±0.2°和16.1°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述L-酒石酸盐Form 1的XRPD图谱还在19.8°±0.2°、13.3°±0.2°、17.0°±0.2°和15.3°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述L-酒石酸盐Form 1具有基本上与图21相同的XRPD图谱。
    优选地，所述L-酒石酸盐Form 1为水合物。
  15. 根据权利要求14所述的L-酒石酸盐Form 1的制备方法，所述制备方法包括以下方法中的任一种：
    1)步骤A:将适量式(I)化合物在溶剂15中溶解；
    步骤B:将适量L-酒石酸在溶剂16中溶解；
    步骤C:将上述两种溶液合并，搅拌析出固体形成混悬液；
    步骤D:继续搅拌或在混悬液中加入溶剂17后继续搅拌，析晶，离心，干燥，得到L-酒石酸盐Form 1。
    优选地，所述溶剂15选自苯环类。
    优选地，所述溶剂16选自醇类。
    优选地，所述溶剂17选自异丙醚。
    优选地，所述步骤D中的搅拌温度为低温，所述低温≤30℃；更优选为0℃-20℃；
    优选地，所述式(I)化合物和L-酒石酸的投料摩尔比为1：1-2.5。
    优选地，所述式(I)化合物和溶剂15的质量体积比(mg/ml)≤300:1；优选为≤200:1；进一步优选为≤150:1。
    2)取式(I)化合物L-酒石酸盐在溶剂18中形成悬浮液，搅拌析晶，得到L-酒石酸盐Form 1。
    优选地，所述溶剂18选自酯类、烷烃类、醇类、醚类和水中的任一种或其组合；所述酯类选自乙酸异丙酯；所述烷烃类选自二氯甲烷、正庚烷、正己烷和甲基环己烷中的任一种或其组合；所述醇类选自甲醇和乙醇中的任一种或其组合；所述醚类选自甲基叔丁基醚。
    优选地，所述溶剂18为特定体积比的混合溶剂体系，比如乙酸异丙酯和正庚烷的混合溶剂，其体积比为1:4；或二氯甲烷和正己烷的混合溶剂，其体积比为1:4；或乙醇和正庚烷的混合溶剂，其体积比为1:4；或甲醇和水的混合溶剂，其体积比为1：2；或二氯甲烷和甲基环己烷的混合溶剂，其体积比为1:6；或甲基叔丁基醚和乙醇的混合溶剂，其体积比为4:1；或乙酸异丙酯和正己烷的混合溶剂，其体积比为1:6。
    优选地，所述搅拌温度为低温、室温或高温。
    优选地，所述低温≤30℃；更优选为0℃-20℃。
    优选地，所述低温为4℃-8℃。
    优选地，所述高温≥30℃；更优选为40℃-60℃。
  16. 根据权利要求13所述的式(I)化合物的盐，其特征在于，所述式(I)化合物L-酒石酸盐为L-酒石酸盐的Form 2，其特征在于，使用Cu-Kα辐射，所述L-酒石酸盐Form2具有基本上与图26相同的XRPD图谱。
    优选地，所述L-酒石酸盐Form 2为水合物。
  17. 根据权利要求13所述的式(I)化合物的盐，其特征在于，所述式(I)化合物L-酒石酸盐为L-酒石酸盐的Form 3，其特征在于，使用Cu-Kα辐射，所述L-酒石酸盐Form 3具有基本上与图30相同的XRPD图谱。
    优选地，所述L-酒石酸盐Form 3为水合物。
  18. 根据权利要求13所述的式(I)化合物的盐，其特征在于，所述式(I)化合物L-酒石酸盐为L-酒石酸盐的Form 4，其特征在于，使用Cu-Kα辐射，所述L-酒石酸盐Form 4以2θ角度表示的XRPD图谱在3.6°±0.2°、11.0°±0.2°和14.3°±0.2°中的至少一处具有特征峰；优选为二处或三处有特征峰。
    优选地，所述L-酒石酸盐Form 4的XRPD图谱还在6.6°±0.2°、7.3°±0.2°、8.4°±0.2°、17.7°±0.2°和18.0°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述L-酒石酸盐Form 4的XRPD图谱还在13.2°±0.2°、13.8°±0.2°、18.5°±0.2°19.9°±0.2°和21.8°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述L-酒石酸盐Form 4具有基本上与图34相同的XRPD图谱。
    优选地，所述L-酒石酸盐Form 4为无水物或通道水合物。
  19. 根据权利要求18所述的L-酒石酸盐Form 4的制备方法，所述制备方法包括以下方法中的任一种：
    1)将L-酒石酸盐Form 1在溶剂20中溶清，然后缓慢挥发析晶，分离晶体，得到L-酒石酸盐Form 4。
    优选地，所述溶剂20选自酮类中、酯类、醚类和苯环类的任一种或其混合；更优选地，所述酮类选自丙酮和甲基异丁酮中的任一种或其组合；所述酯类选自乙酸乙酯和乙酸异丙酯中的任一种或其组合；所述醚类选自甲基叔丁基醚；所述苯环类选自甲苯。
    优选地，所述溶剂20为单一溶剂体系，比如乙酸异丙酯。
    优选地，所述溶剂20为特定体积比的混合溶剂体系，比如丙酮和甲基叔丁基醚的混合溶剂，其体积比为1:1；或者乙酸乙酯和甲苯的混合溶剂体系，其体积比为2:1。
    优选地，所述挥发的时间≥0.5天。
    优选地，所述缓慢挥发为小孔挥发。
    优选地，所述缓慢挥发的温度为室温。
    2)将L-酒石酸盐Form 1在溶剂21中形成悬浮液，加入L-酒石酸盐Form 4晶种，室温搅拌析出，分离晶体，得到L-酒石酸盐Form 4。
    优选地，所述溶剂21为酮类、烷烃类、酯类和醚类中的任一种或其组合；更优选地，所述酮类选自丙酮和甲基异丁酮中的任一种或其组合；所述烷烃类选自正庚烷；所述酯类选自乙酸乙酯；所述醚类选自甲基叔丁基醚。
    优选地，所述溶剂21为特定体积比的混合溶剂体系，比如丙酮和正庚烷的混合溶剂，其体积比为1:6；或乙酸乙酯和正庚烷的混合溶剂，其体积比为1:6；或乙酸异丙酯和正庚烷的混合溶剂，其体积比为1:4，或丙酮和甲基叔丁基醚的混合溶剂，其体积比为1:4。
    3)将适量式(I)化合物和适量L-酒石酸在溶剂22中形成悬浮液，加入L-酒石酸盐Form 4晶种，搅拌析出，分离晶体，得L-酒石酸盐Form 4。
    优选地，所述溶剂22选自为酮类、烷烃类、酯类和醚类中的任一种或其组合；更优选地，所述酮类选自丙酮和甲基异丁酮中的任一种或其组合；所述烷烃类选自正庚烷；所述酯类选自乙酸乙酯；所述醚类选自甲基叔丁基醚。
    优选地，所述溶剂22为特定体积比的混合溶剂体系，比如丙酮和正庚烷的混合溶剂，其体积比为1:6。
    4)取式(I)化合物L-酒石酸盐，在溶剂23中形成悬浮液，搅拌析晶，得到L-酒石酸盐Form 4。
    优选地，所述溶剂23选自烷烃类、酮类和醚类中的任一种或其组合；更优选地，所述烷烃类选自正庚烷；所述酮类选自丁酮；所述醚类选自甲基叔丁基醚。
    优选地，所述溶剂23为甲基叔丁基醚。
    优选地，所述溶剂23为特定体积比的混合溶剂体系，比如丁酮和正庚烷的混合溶剂，其体积比为1:6。
    优选地，所述搅拌温度为低温或室温。
    优选地，所述低温≤30℃；更优选为0℃-20℃。
    在一些实施例中，所述低温为4℃-8℃。
  20. 根据权利要求13所述的式(I)化合物的盐，其特征在于，所述式(I)化合物L-酒石酸盐为L-酒石酸盐Form 5，其特征在于，使用Cu-Kα辐射，所述L-酒石酸盐Form5具有基本上与图38相同的XRPD图谱。
  21. 根据权利要求13所述的式(I)化合物的盐，其特征在于，所述式(I)化合物L-酒石酸盐为L-酒石酸盐Form 6，其特征在于，使用Cu-Kα辐射，所述L-酒石酸Form 6具有基本上与图39相同的XRPD图谱。
  22. 根据权利要求13所述的式(I)化合物的盐，其特征在于，所述式(I)化合物L-酒石酸盐为L-酒石酸盐无定型，其特征在于，使用Cu-Kα辐射，所述L-酒石酸盐无定型以2θ表示的XRPD图谱中无尖锐的衍射峰。
    优选地，所述L-酒石酸盐无定型具有基本上与40相同的XRPD图谱。
  23. [根据细则26改正 11.04.2024]
    根据权利要求1或2所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐选自盐酸盐，结构式如式(IV)所示：
    优选地，所述式(I)化合物的盐酸盐基本上以结晶形式或无定型形式存在。
  24. 根据权利要求23所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐酸盐为盐酸盐Form 1，其特征在于，使用Cu-Kα辐射，所述盐酸盐Form 1以2θ角度表示的XRPD图谱13.5°±0.2°、15.4°±0.2°、15.7°±0.2°、32.6°±0.2°和35.3°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处或五处有特征峰。
    优选地，所述盐酸盐Form 1具有基本上与图45相同的XRPD图谱。
  25. 根据权利要求1或2所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐选自甲磺酸盐，结构式如式(V)所示：
    优选地，所述式(I)化合物的甲磺酸盐基本上以结晶形式或无定型形式存在。
  26. 根据权利要求25所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的甲磺酸盐为甲磺酸盐的Form 1，其特征在于，使用Cu-Kα辐射，所述甲磺酸盐Form1具有基本上与图49相同的XRPD图谱。
  27. 根据权利要求25所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的甲磺酸盐为甲磺酸盐Form 2，其特征在于，使用Cu-Kα辐射，所述甲磺酸盐Form 2以2θ角度表示的XRPD图谱在6.5°±0.2°、13.8°±0.2°、15.2°±0.2°和17.0°±0.2°中的至少一处具有特征峰；优选为二处或三处或四处有特征峰。
    优选地，所述甲磺酸盐Form 2具有基本上与图50相同的XRPD图谱。
  28. 根据权利要求25所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的甲磺酸盐为甲磺酸盐Form 3，其特征在于，使用Cu-Kα辐射，所述甲磺酸盐Form 3具有基本上与图54相同的XRPD图谱。
  29. 根据权利要求1或2所述的式(I)化合物的盐，其特征在于，所述式(I)化合物的盐选自草酸盐，结构式如式(VI)所示:
    优选地，所述式(I)化合物草酸盐基本上以结晶形式或无定型形式存在。
  30. 根据权利要求29所述的式(I)化合物的盐，其特征在于，所述式(I)化合物草酸盐为草酸盐Form 1，其特征在于,使用Cu-Kα辐射，所述草酸盐Form 1具有基本上与图55相同的XRPD图谱。
  31. 根据权利要求29所述的式(I)化合物的盐，其特征在于，所述式(I)化合物草酸盐为草酸盐Form 2，其特征在于，使用Cu-Kα辐射，所述草酸盐Form 2以2θ角度表示的XRPD图谱在15.5°±0.2°、17.2°±0.2°和23.8°±0.2°中的至少一处具有特征峰；优选为二处或三处有特征峰。
    优选地，所述草酸盐Form 2的XRPD图谱还在13.9°±0.2°、16.9°±0.2°、19.8°±0.2°和16.0°±0.2°2θ中的至少一处具有特征峰。
    优选地，所述草酸盐Form 2具有基本上与图56相同的XRPD图谱。
  32. 一种药物组合物，所述药物组合物包含治疗有效量的权利要求1-31中任一项所述的式(I)化合物的盐或其晶型，以及至少一种药学上可接受的载体。
  33. 一种权利要求1-31中任一项所述的式(I)化合物的盐或其晶型、或其药物组合物在制备预防和/或治疗STAT3双功能磷酸化位点活化引起或调控的疾病的药物中的应用。
    优选地，所述应用为抑制癌细胞的增殖、生长、迁移、浸润、克隆形成和转移，促进癌细胞的凋亡，和/或延长肿瘤患者的生存期。
    优选地，所述疾病选自胰腺癌Capan-2、PANC-1、MIAPACA-2、BXPC3、SW1990、CFPAC-1、Pan02、ASPC-1，胃癌MKN45、BGC823、MGC803，肝癌Huh7、PLC/PRF-5、HepG2、SK-hep1、SMMC7721、HCCC9810、肺癌HCC827、A549、H460、H23、H1299、H522、H1975、PC-9，前列腺癌DU145、LNCAP，结肠癌HCT8、HT29、HCT-116。
  34. 一种权利要求1-31中任一项所述的式(I)化合物的盐或其晶型、或其药物组合物在制备抑制STAT3的磷酸化、抑制STAT3的转录活性和线粒体的氧化磷酸化的抑制剂中的应用。
  35. 一种预防和/或治疗与STAT3双功能磷酸化位点活化有关的疾病的方法，其包括向有需要的个体中施于有效量的权利要求1-31中任一项所述的式(I)化合物的盐或其晶型、或其药物组合物。
    优选地，所述疾病选自胰腺癌Capan-2、PANC-1、MIAPACA-2、BXPC3、SW1990、CFPAC-1、Pan02、ASPC-1、胃癌MKN45、BGC823、MGC803，肝癌Huh7、PLC/PRF-5、HepG2、SK-hep1、SMMC7721、HCCC9810，肺癌HCC827、A549、H460、H23、H1299、H522、H1975、PC-9，前列腺癌DU145、LNCAP，结肠癌HCT8、HT29和HCT-116。
  36. 一种权利要求1-31中任一项所述的式(I)化合物的盐或其晶型其药物组合物与其他药物的联合应用。
PCT/CN2024/083481 2023-03-27 2024-03-25 三芳香环类衍生物的盐、其晶型、制备方法和用途 Pending WO2024199167A1 (zh)

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