Salt of 3, 4-dihydroisoquinoline compound as well as preparation method and application thereof

Abstract

The invention provides hippurate of a compound as shown in a formula (A), a solid form, a crystal form and a specific crystal form of the hippurate, a pharmaceutical composition containing the hippurate and application of the hippurate. The hippurate of the compound shown in the formula (A) has the characteristics of good solubility, low hygroscopicity, good stability and the like, and is suitable for being used as a raw material medicine. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and relates to hippurate of 3,4-dihydroisoquinoline compounds, solid forms, crystalline forms and specific crystal forms thereof, preparation methods thereof, pharmaceutical compositions containing the same and applications thereof in the medical field. Background Art

[0002] Protein arginine methyltransferases (PRMTs) are a class of S-adenosylmethionine (SAM or AdoMet)-dependent methyltransferases that can catalyze protein arginine methylation reactions. Specifically, they are responsible for transferring the methyl group from AdoMet to the guanidinium nitrogen atom at the end of the arginine residue of histones or other proteins. PRMTs play an important role in protein methylation, such as participating in alternative splicing, post-transcriptional regulation, RNA processing, cell proliferation, cell differentiation, apoptosis and tumor formation. According to the different ways of catalyzing arginine methylation, members of the PRMTs family can be divided into three categories: PRMT1-4, PRMT6, and PRMT8 belong to type I, catalyzing monomethyl and asymmetric dimethyl; PRMT5 and PRMT9 belong to type II, catalyzing symmetric dimethyl; PRMT7 belongs to type III and can catalyze monomethyl.

[0003] PRMT5 was first isolated from a protein complex that binds to Jak2 (Janus tyrosinekinase 2) by Pollack et al. in yeast two-hybrid studies, so it is also called JBP1 (jak-binding protein 1). PRMT5 can not only regulate the process of gene transcription and protein modification, but also regulate cell proliferation, differentiation, and apoptosis during the growth of tumor cells. Overexpression of PRMT5 is found in many tumors or cancers, and it is a very potential target for tumor treatment. For example, studies have shown that PRMT5 is upregulated in solid tumors (e.g., lung cancer, bone cancer, gastric cancer, pancreatic cancer, adenoid cystic carcinoma, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, brain cancer, pituitary adenoma, melanoma, epidermoid carcinoma) and blood tumors (e.g., lymphoma, leukemia, chronic and acute leukemia, acute myeloid leukemia). So far, the research and development of PRMT5 inhibitors is in the early stages. GSK3326595 released by GSK has the highest research and development status of Phase II clinical trials. JNJ-64619178 developed by Janssen, PF-06939999 developed by Pfizer, and PRT-543 developed by Prelude Therapeutics are all in Phase I clinical trials.

[0004] WO2021244542A1 discloses a series of PRMT5 inhibitors with novel structures, including compound PR-4 (the compound shown in the following formula (A)).

[0005] Summary of the invention

[0006] WO2021244542A1 does not record the physical form of the compound shown in the obtained formula (A). The inventor obtained the compound shown in formula (A) according to the preparation method described in WO2021244542A1 as a colloid (or colloid, oil), and tried various methods, but failed to obtain the solid form of the compound shown in formula (A). Furthermore, the inventor tried to solidify it by salification, using a variety of common acids, but did not obtain a solid or the resulting solid was unstable. Unexpectedly, the inventor found a hippurate salt of the compound shown in formula (A), which is not only in solid form, but also has good properties and is suitable for use as a bulk drug.

[0007] In a first aspect, the present invention provides a compound, which is a hippurate salt of a compound represented by formula (A),

[0008]

[0009] According to some embodiments of the present invention, the compound is a compound represented by formula (A-1),

[0010]

[0011] Among them, n is selected from 0.5-2; preferably 1-1.5; more preferably 1, 1.1, 1.2, 1.3, 1.4 or 1.5; more preferably 1, 1.3, 1.4 or 1.5; further preferably 1.

[0012] According to some embodiments of the present invention, the compound is a compound represented by formula (B):

[0013]

[0014] According to some embodiments of the present invention, the compound, the compound represented by formula (A-1) or the compound represented by formula (B) is in solid form.

[0015] According to some embodiments of the present invention, the compound of formula (B) in solid form, using the KBr tablet method, has an infrared spectrum including characteristic peaks at the following positions (±4 cm -1): 3281, 2939, 1653, 1524, 1373, 1305; preferably, the KBr tablet method is used, and its infrared spectrum includes characteristic peaks at the following positions (±4 cm -1 ): 3281, 2939, 1653, 1524, 1373, 1305, 1121, 1038, 750; further preferably, using the KBr tablet method, its infrared spectrum includes characteristic peaks at the following positions (±4cm -1 ): 3281, 2939, 1653, 1612, 1524, 1457, 1373, 1305, 1147, 1121, 1038, 976, 750, 690; further preferably, using the KBr tablet method, its infrared spectrum is substantially as follows Figure 1 shown.

[0016] According to some embodiments of the present invention, the compound, the compound represented by formula (A-1) or the compound represented by formula (B) is in crystalline form.

[0017] According to some embodiments of the present invention, the compound represented by formula (B), which is the crystalline form I of the compound represented by formula (B), uses Cu-Kα radiation, and the X-ray powder diffraction pattern expressed in 2θ angles comprises the following characteristic diffraction peaks (±0.3°, preferably ±0.2°): 6.1, 16.0, 18.4, 20.1, 21.7.

[0018] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) has an X-ray powder diffraction pattern expressed at an angle of 2θ using Cu-Kα radiation, comprising the following characteristic diffraction peaks (±0.3°, preferably ±0.2°): 6.1, 13.3, 16.0, 18.4, 20.1, 21.7.

[0019] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) has an X-ray powder diffraction pattern expressed at an angle of 2θ using Cu-Kα radiation, comprising the following characteristic diffraction peaks (±0.3°, preferably ±0.2°): 6.1, 8.1, 13.3, 16.0, 18.4, 20.1, 21.7.

[0020] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) has an X-ray powder diffraction pattern expressed at an angle of 2θ using Cu-Kα radiation, comprising the following characteristic diffraction peaks (±0.3°, preferably ±0.2°): 6.1, 8.1, 13.3, 15.0, 16.0, 18.4, 20.1, 21.7.

[0021] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) has an X-ray powder diffraction pattern expressed at an angle of 2θ using Cu-Kα radiation, comprising the following characteristic diffraction peaks (±0.3°, preferably ±0.2°): 6.1, 8.1, 13.3, 15.0, 16.0, 17.8, 18.4, 20.1, and 21.7.

[0022] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) has an X-ray powder diffraction pattern expressed at an angle of 2θ using Cu-Kα radiation, comprising the following characteristic diffraction peaks (±0.3°, preferably ±0.2°): 4.9, 6.1, 8.1, 13.3, 15.0, 16.0, 17.8, 18.4, 20.1, 21.7.

[0023] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) has substantially the following 2θ angles when using Cu-Kα radiation: Figure 2 or Figure 4 The X-ray powder diffraction pattern is shown.

[0024] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) has an endothermic peak at 167°C±5°C in its differential scanning calorimetry curve.

[0025] In a second aspect, the present invention provides a crystalline composition comprising Form I of the compound represented by formula (B).

[0026] According to some embodiments of the present invention, the crystalline form I of the compound represented by formula (B) accounts for more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more than 95% of the weight of the crystalline composition.

[0027] In a third aspect, the present invention provides a pharmaceutical composition comprising the hippurate salt of the compound of formula (A), the compound of formula (A-1), the compound of formula (B), the crystalline form I of the compound of formula (B), or the crystalline composition described in the second aspect, optionally further comprising a pharmaceutically acceptable carrier.

[0028] In a fourth aspect, the present invention provides the use of the hippurate salt of the compound of formula (A) described in the first aspect, the compound of formula (A-1), the compound of formula (B), the crystalline form I of the compound of formula (B), the crystalline composition described in the second aspect, or the pharmaceutical composition described in the third aspect in the preparation of drugs.

[0029] According to some embodiments of the present invention, the drug is used to prepare a drug for preventing and / or treating a cell proliferative disease; preferably, the cell proliferative disease is a tumor or cancer; further preferably, the tumor or cancer is a blood tumor or a solid tumor; further preferably, it is a malignant blood tumor or an advanced solid tumor; further preferably, it is a relapsed refractory blood tumor or an advanced malignant solid tumor.

[0030] According to some embodiments of the present invention, the drug is for preparing a drug for preventing and / or treating a disease at least partially mediated by PRMT5.

[0031] According to some embodiments of the present invention, the disease at least partially mediated by PRMT5 is a cell proliferative disease; preferably, the cell proliferative disease is a tumor or cancer; further preferably, the tumor or cancer is a blood tumor or a solid tumor; further preferably, it is a malignant blood tumor or an advanced solid tumor; further preferably, it is a relapsed refractory blood tumor or an advanced malignant solid tumor.

[0032] According to some embodiments of the present invention, the cell proliferative disease is a tumor or cancer; preferably, the tumor or cancer is a blood tumor or a solid tumor; further preferably, it is a malignant blood tumor or an advanced solid tumor; further preferably, it is a relapsed refractory blood tumor or an advanced malignant solid tumor.

[0033] According to some embodiments of the present invention, the tumor or cancer is selected from lung cancer, bone cancer, gastric cancer, pancreatic cancer, adenoid cystic carcinoma, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, brain cancer, pituitary adenoma, melanoma, epidermoid carcinoma, lymphoma, leukemia, and chronic and acute leukemia; preferably, the acute leukemia is acute myeloid leukemia (AML).

[0034] In a fifth aspect, the present invention provides a method for preparing a hippurate salt of a compound represented by formula (A), comprising reacting the compound represented by formula (A) with hippuric acid in a reaction solvent, and separating and obtaining a hippurate salt of the compound represented by formula (A).

[0035] In some embodiments, the present invention provides a method for preparing a compound represented by formula (A-1), comprising reacting a compound represented by formula (A) with an acid in a solvent to obtain a compound represented by formula (A-1):

[0036]

[0037] Wherein, X is hippuric acid; n is selected from 0.5-2.

[0038] According to some embodiments of the present invention, n is selected from 1-1.5; preferably 1, 1.1, 1.2, 1.3, 1.4 or 1.5; further preferably 1, 1.3, 1.4 or 1.5; and further preferably 1.

[0039] According to the preparation method of the present invention, the molar ratio of the compound represented by formula (A) to the acid is 1-2:0.5-2, preferably 1:1-2, and more preferably 1:1.1-2, for example, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:2.

[0040] According to the preparation method of the present invention, the reaction temperature is 0°C to 90°C, preferably 5°C to 80°C, preferably 20°C to 60°C, more preferably room temperature to 50°C.

[0041] According to the preparation method of the present invention, the solvent is selected from one or a combination of two of methanol, ethanol, dichloromethane, and methyl tert-butyl ether; preferably, it is a single solvent of methanol, ethanol or dichloromethane or a mixed solvent of methanol and methyl tert-butyl ether; further preferably, it is ethanol; preferably, when it is a mixed solvent of methanol and methyl tert-butyl ether, the volume ratio of the two is 1-20:20-1, preferably 1-19:19-1, preferably 1-10:10-1, preferably 1-5:5-1, preferably 3:5.

[0042] According to the preparation method of the present invention, after the reaction is completed, the temperature is further reduced to -15°C to room temperature; preferably -10°C to room temperature; further preferably -5°C to room temperature; for example, -5°C to 5°C (0±5°C), 0°C to 10°C (5±5°C).

[0043] According to the preparation method of the present invention, after the reaction is completed, a crystallization step is further included, preferably stirring crystallization or static crystallization.

[0044] According to the preparation method of the present invention, the reaction is completed, and the static crystallization is selected from solvent evaporation crystallization or solvent in-crystallization.

[0045] According to the preparation method of the present invention, the reaction is completed, and crystallization is carried out for 0.5 hours to 10 days; preferably 0.5 hours to 8 days; 0.5 hours to 6 days; 0.5 hours to 5 days; 0.5 hours to 4 days; 0.5 hours to 3 days; 0.5 hours to 2 days; 0.5 hours to 24 hours; 0.5 hours to 12 hours; for example, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours. According to the preparation method of the present invention, the reaction is completed, and further includes a separation step, which is any suitable method, and the separation is to obtain a solid. Preferably, the separation is selected from suction filtration, centrifugation, filtration, and suction filtration.

[0046] The preparation method according to the present invention further comprises a drying step to obtain a compound represented by formula (A-1).

[0047] According to the preparation method of the present invention, the drying method can adopt any suitable known method, preferably room temperature drying, room temperature vacuum drying, drying at 50°C ± 5°C, drying at 60°C ± 5°C or drying at 65°C ± 5°C. Specific drying conditions are, for example, the drying time is preferably 1h to 5 days, more preferably 3h to 3 days, more preferably 3h to 1 day, for example, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours. Regardless of the drying means, it is advisable that the residual solvent in the obtained product meets the quality standards.

[0048] The compound represented by formula (A) mentioned herein is prepared by the method disclosed in WO2021244542A1, and can also be prepared by any known method disclosed in other prior arts or a combination thereof, or by replacing, combining or improving the method disclosed in WO2021244542A1 and / or other prior arts.

[0049] Definition and Description

[0050] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be considered ambiguous or unclear without special definition, but should be understood according to the ordinary meaning.

[0051] The compounds of formula (A), formula (A-1) and / or formula (B) mentioned in the present invention may exist in specific stereoisomer forms, including cis and trans isomers, (R)- and (S)-enantiomers, and racemic mixtures thereof. The compounds of the present invention may contain a carbon-nitrogen double bond in E or Z configuration, wherein the term "E" represents a higher order substituent on the opposite side of the carbon-nitrogen double bond, and the term "Z" represents a higher order substituent on the same side of the carbon-nitrogen double bond (determined using the Cahn-Ingold Prelog priority rule). The compounds of the present invention may also exist in the form of a mixture of "E" and "Z" isomers. Preferably, the compounds of the present invention exist in the form of E isomers. The substituents around the heterocycloalkyl group are made into cis or trans configurations.

[0052] The "solid form" mentioned in the present invention refers to a compound in solid form, including but not limited to a crystalline form and an amorphous form of the compound.

[0053] The "crystalline form" mentioned in the present invention refers to a compound in a crystalline form, including but not limited to anhydrous and solvent-free forms, hydrate forms and solvate forms.

[0054] The term "solvate" also known as "solvate" refers to an association formed by a stoichiometric or non-stoichiometric ratio of solvent molecules and the compounds of the present invention, including an association containing both water molecules and one or more other solvent molecules, and an association containing only one or more other solvent molecules.

[0055] The term "hydrate" refers to an association formed between a stoichiometric or non-stoichiometric ratio of water molecules and a compound of the present invention.

[0056] The "anhydrous and solvent-free form" refers to the absence of water molecules or solvent molecules, or the coexistence of water molecules or solvent molecules with the compound of the present invention in a manner that is not bound by intermolecular forces, such as by adsorption.

[0057] The term "crystalline composition" refers to a solid form that contains one or more of the specific crystal forms of the compound of the present invention. For example, in one embodiment according to the present invention, it contains the crystal form I of the compound of formula (B) mentioned in the present invention. Moreover, in addition to the crystal form of the present invention, the crystalline composition may optionally contain other crystalline forms, other crystal forms or other amorphous forms of the compound of the present invention, or impurities other than these substances. It should be understood by those skilled in the art that the sum of the contents of each component in the crystalline composition should be 100%.

[0058] The "room temperature" is the room temperature conventionally known in the art, generally 10 to 30°C, preferably 25°C±5°C.

[0059] In the context of the present invention, the diffraction angles in an X-ray powder diffraction pattern are expressed in degrees 2θ, with the unit being degrees (°).

[0060] In the X-ray powder diffraction pattern, the term "substantially" or "substantially as shown" refers to a substantially pure crystalline form, in which at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% of the peaks in the powder X-ray diffraction pattern appear in the given pattern. Further, when the content of a certain crystalline form in the product gradually decreases, some of the diffraction peaks attributable to the crystalline form in its X-ray powder diffraction pattern may become less due to the detection sensitivity of the instrument. In addition, for any given crystalline form, there may be slight errors in the position of the peak, which is also well known in the field of crystallography. For example, due to changes in temperature, sample movement or instrument calibration during sample analysis, the position of the peak may move, and there is a certain measurement error in the 2θ value. Therefore, when determining each crystalline structure, this error should be taken into account, and the term "substantially" or "substantially as shown in the accompanying drawings" is also intended to cover such differences in the diffraction peak positions, referring to ±0.3°, preferably ±0.2°.

[0061] In the DSC spectrum, the term "substantially" or "substantially as shown" means that for the same crystal form of the same compound, in consecutive analyses, the error of the thermal transition onset temperature, the peak temperature of the endothermic peak, the peak temperature of the exothermic peak, the melting point, the starting temperature of weight loss or the end temperature of weight loss is typically within about 5°C, usually within about 3°C. When describing a compound as having a given thermal transition onset temperature, peak temperature of the endothermic peak, peak temperature of the exothermic peak, the melting point, the starting temperature of weight loss or the end temperature of weight loss, it refers to the temperature ±5°C.

[0062] As used herein, the term "cell proliferative disorder" refers to a condition in which a population of cells grows at a rate that is slower or faster than expected under given physiological conditions and circumstances.

[0063] The term "tumor" includes benign tumors, malignant tumors and borderline tumors, among which malignant tumors are collectively referred to as cancer.

[0064] As used herein, the term "prevent" or "prevent" means that when used for a disease or condition (e.g., cancer), the compound or drug (e.g., the combination product claimed herein) can reduce the frequency of symptoms of the medical condition or delay its onset in a subject compared to a subject to which the compound or drug (e.g., the combination product claimed herein) is not administered.

[0065] As used herein, the term "treat," "treat," "treat," "treat," or "treat" refers to alleviating, relieving, or ameliorating symptoms of a disease or condition, ameliorating underlying metabolically caused symptoms, inhibiting a disease or symptom, such as arresting the development of a disease or condition, alleviating a disease or condition, causing regression of a disease or condition, alleviating a condition caused by a disease or condition, or arresting symptoms of a disease or condition.

[0066] The term "pharmaceutically acceptable carrier" may also be referred to as "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient", which refers to those carriers or excipients that have no obvious irritation to the organism and will not impair the biological activity and performance of the active compound.

[0067] The "jelly" mentioned herein is also called "colloid" or "oil", which is in a state similar to oil and is not in a solid form.

[0068] The above embodiments represent exemplary embodiments of the present invention, but the present invention is not limited to the above embodiments. In addition, the various technical features in the above embodiments of the present invention can be combined with each other to form one or more new technical solutions, which also fall within the scope of the present invention, as long as such new technical solutions are technically feasible.

[0069] The term "optionally" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or not.

[0070] The intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention.

[0071] The chemical reactions of the specific embodiments of the present invention are carried out in a suitable solvent, which must be suitable for the chemical changes of the present invention and the reagents and materials required. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments.

[0072] The present invention will be described in detail below by way of examples, which are not intended to limit the present invention in any way.

[0073] All solvents used in the present invention were commercially available and used without further purification.

[0074] Technical Effects

[0075] The hippurate salt of the compound represented by formula (A) provided by the present invention, its solid form, crystalline form and crystal form thereof have one or more of the following beneficial effects:

[0076] The hippurate salt of the compound represented by formula (A) was unexpectedly obtained for the first time and can be prepared and isolated with a relatively high purity (greater than 95%);

[0077] The hippurate salt of the compound represented by formula (A) is obtained for the first time in solid form, preferably, the hippurate salt of the compound represented by formula (A) is obtained for the first time in crystalline form, which is convenient for separation, transfer, weighing and formulation process operation;

[0078] The crystalline form of the compound represented by formula (B) is obtained for the first time, which has good crystallinity and is convenient for separation, transfer, weighing and formulation process operation;

[0079] The crystalline form I of the compound represented by formula (B) is obtained for the first time, which has good properties and good crystallinity;

[0080] The crystalline form I of the compound represented by formula (B) has good solubility in water and biological solvents;

[0081] Compared with other reference products, the crystalline form I of the compound represented by formula (B) has no obvious hygroscopicity;

[0082] The crystalline form I of the compound represented by formula (B) has good physical stability;

[0083] The compound represented by formula (B) and its crystalline form I have good chemical stability;

[0084] The tablet (tablet core) prepared using the crystalline form I of the compound represented by formula (B) has a good dissolution effect, preferably, can dissolve quickly. BRIEF DESCRIPTION OF THE DRAWINGS

[0085] Figure 1 : Infrared spectrum of the compound of formula (B) of Example 1.

[0086] Figure 2 : X-ray powder diffraction pattern of Form I of the compound of formula (B) of Example 1.

[0087] Figure 3 : Differential scanning calorimetry spectrum of Form I of the compound of formula (B) of Example 1.

[0088] Figure 4 : X-ray powder diffraction pattern of the sample in Example 1 after stability testing. DETAILED DESCRIPTION

[0089] 1. X-ray powder diffractometer (XRPD)

[0090]

[0091]

[0092] 2. Differential Scanning Calorimeter (DSC)

[0093]

[0094] 3. Nuclear Magnetic Resonance Spectroscopy (NMR) Instrument model: Bruker 400M NMR instrument (Bruker, GER)

[0095] Content and test solvent: 1 H-NMR, the test solvent was DMSO-d6.

[0096] 4. High Performance Liquid Chromatography (HPLC)

[0097] 4.1 Detection instrument: Waters H-Class UPLC (ultra-high performance liquid chromatography)

[0098]

[0099] 4.2 Detection instrument: Waters e2695 (high performance liquid chromatograph)

[0100]

[0101] 5. Moisture absorption test

[0102] Description of hygroscopic characteristics and definition of hygroscopic weight gain:

[0103] Moisture-attracting properties Weight gain ratio Highly hygroscopic The weight gain due to moisture absorption is not less than 15% Hygroscopic The weight gain due to moisture absorption is less than 15% but not less than 2% Slightly hygroscopic The weight gain due to moisture absorption is less than 2% but not less than 0.2% No or almost no hygroscopicity The weight gain due to moisture absorption is less than 0.2%

[0104] 5.1 Dynamic moisture sorption (DVS)

[0105] Instrument Model SMS(Surface Measurement Systems) sample Example 1 Sample volume 20–40 mg Shielding gas and flow rate Nitrogen, 200mL / min dm / dt 0.002% / min

[0106] 5.2 Moisture absorption and weight gain test

[0107] Sample: Preparation Example 1

[0108] The method in the Chinese Pharmacopoeia was used for determination. The specific test method is as follows:

[0109] 1) Take a dry stoppered glass weighing bottle (outer diameter 50mm, height 15mm), place it in a suitable 25℃±1℃ constant temperature drying oven (with saturated ammonium chloride solution placed at the bottom) one day before the test, and accurately weigh the weight (m1);

[0110] 2) Take an appropriate amount of the test sample and spread it flat in the above-mentioned weighing bottle. The thickness of the test sample is generally about 1mm. Accurately weigh the weight (m2);

[0111] 3) Open the weighing bottle and place it under the above constant temperature and humidity conditions with the bottle cap on for 24 hours;

[0112] 4) Cover the weighing bottle and accurately weigh the weight (m3);

[0113] Percentage of weight gain = (m3-m2) / (m2-m1)×100%.

[0114] 6. Moisture determination

[0115] Instrument Model 915KF Ti-Touch Test samples Preparation Example 1 Sample quantity 100 mg Solvents Anhydrous methanol Karl Fischer reagent KFR-08

[0116] 7. Infrared Spectroscopy (IR)

[0117] Detection instrument: PerkinElmer Spectrum 100 infrared spectrometer

[0118] Test method: Weigh 3 mg of sample, dilute with KBr and press into a tablet, and test at room temperature. The specific parameters are: Detection range: 4000-400 cm -1 Wave number, resolution: 4 cm -1 .

[0119] In order to better understand the content of the present invention, the following is further described in conjunction with specific examples, but the specific implementation methods are not intended to limit the content of the present invention. The test methods without specific conditions in the following preparation examples, embodiments, comparative examples or test examples are selected according to conventional methods and conditions, or according to the product instructions.

[0120] Preparation Example 1: Preparation of the compound of formula (A)

[0121] The compound represented by formula (A) was prepared by referring to the method disclosed in WO2021244542A1 to obtain the compound represented by formula (A) as a colloid.

[0122]

[0123] Example 1: Preparation of compound of formula (B)

[0124] Add the sample of Preparation Example 1 (18.38 g), anhydrous ethanol (550 mL) and hippuric acid (6.47 g) to the reaction bottle in sequence and stir to dissolve. Continue stirring overnight to precipitate a large amount of solid. Cool the reaction bottle to 0±5°C and continue stirring to crystallize for 2 hours. Filter and vacuum dry at 50±5°C for 5 hours to obtain a solid (20.2 g). Detection by nuclear magnetic hydrogen spectrum confirmed that salt had been formed and the base / acid ratio was 1:1.

[0125] Take samples for infrared spectrum test, IR (KBr, cm -1 ):3281.34, 2939.10, 1652.92, 1612.15, 1524.15, 1456.64, 1373.01, 1305.15, 1147.29, 1121.18, 1037.56, 975.67, 750.23, 689.66. See the spectrum Figure 1 .

[0126] The sample was taken for X-ray powder diffraction, which showed a crystalline solid (form I) with good crystallinity. Figure 2 The main XRPD diffraction peak data are shown in the table below. The sample was tested by DSC. The DSC graph showed an endothermic peak at 168.77℃. Figure 3 .

[0127] Table 1. XRPD diffraction peak data table of the sample obtained in Example 1

[0128]

[0129] Example 2-4: Crystal form screening

[0130] The samples of Example 1 (about 100 mg) were weighed in sample bottles, and solvents were added and the operations were performed according to the following table. The obtained samples were subjected to X-ray powder diffraction test and DSC test. The results are shown in the following table:

[0131] Table 2. Crystal form screening methods and results

[0132]

[0133] Note: Drying conditions are: 50±5℃, vacuum drying for 5 hours.

[0134] Comparative Example

[0135] During the research and development process, the inventors found that: (1) the compound represented by formula (A) could not obtain a solid sample under various solid research methods and different experimental conditions; (2) after the compound represented by formula (A) reacted with various commonly used acids under different experimental conditions, no solid sample could be obtained, or the obtained solid sample was unstable. Exemplary solutions include but are not limited to the following comparative examples:

[0136] Comparative Example 1: Preparation attempt of the compound of formula (A) in solid form

[0137] The sample obtained in Preparation Example 1 was taken and various physical operations such as reduced pressure rotary evaporation (circulating water pump, water bath at 50°C, 2 to 5 hours), reduced pressure vacuum extraction (oil pump, 3 to 8 hours), or multiple column chromatography purification operations were tried, but none of them could change the properties of the sample.

[0138] Comparative Example 2: Preparation attempt of the compound of formula (A) in solid form

[0139] The samples obtained in Preparation Example 1 were taken and the solvents shown in the following table were added to try crystallization. The specific test methods and results are shown in the following table. In the end, no solid was obtained and the samples were still in the form of colloid.

[0140] Table 3. Preparation conditions and results of the compound of formula (A) in solid form

[0141] Comparative Example Solvents result 1-1 Ethanol Colloid[1] 1-2 Ethyl acetate Colloid[1] 1-3 2-Methyltetrahydrofuran Colloid[1] 1-4 Acetone / water (19:1, v / v) Colloid[1] 1-5 acetone Colloid[1] 1-6 Acetonitrile Colloid[1] 1-7 Tetrahydrofuran Colloid[1] 1-8 Methyl tert-butyl ether Colloid[2] 1-9 Dichloromethane Colloid[1]

[0142] [1]: The solution becomes clear after stirring at room temperature for 3 days. It remains clear after stirring at 5°C for 3 days. After adding the anti-solvent n-hexane or methyl tert-butyl ether, no solid or oil / gel is formed. It evaporates after being transferred to room temperature.

[0143] [2]: After stirring at room temperature for 3 days, if there is still no solid or oil / gel, transfer to room temperature for evaporation.

[0144] Comparative Example 3: Preparation of salts of other compounds of formula (A)

[0145] The samples of Preparation Example 1 were dissolved in the solvents shown in the following table to prepare a solution with a concentration of 40 mg / mL; the sample solution of Preparation Example 1 (0.5 mL) and the corresponding acid ligand in an equal molar ratio were added to a vial. The specific test method and results are shown in the following table:

[0146] Table 4. Preparation and results of other acid addition salts

[0147]

[0148] [1]: The solution becomes clear after stirring at room temperature for 3 days. It remains clear after stirring at 5°C for 3 days. After adding the anti-solvent n-hexane or methyl tert-butyl ether, no solid or oil / gel is formed. It evaporates after being transferred to room temperature.

[0149] [2]: After stirring at room temperature for 3 days, the solution becomes clear or gelled. After stirring at 5°C for 3 days, the solution becomes gelled. After stirring at room temperature, the solution evaporates.

[0150] Results: After the sample of Preparation Example 1 reacted with some common organic acids or inorganic acids, no solid was obtained.

[0151] Comparative Example 4: Preparation of salts of other compounds of formula (A)

[0152] The sample of Preparation Example 1 was reacted with sulfuric acid, phosphoric acid and succinic acid in equal molar ratios in certain solvents to obtain solid samples. However, after the samples were left open for a period of time under ambient conditions, they all turned into colloids or oils. This may be because the solids are unstable or have strong hygroscopicity.

[0153] Test Example 1: Solubility Test

[0154] The samples of Example 1 and Preparation Example 1 were weighed into glass vials, added into different media (water and three biological solvents, with a volume of 4 mL), and solubility tests were performed. The results are shown in the table below.

[0155] Table 5. Solubility of different samples in biological solvents

[0156]

[0157] Note: ① Solubility, measured in free state. ② FaSSIF: simulated intestinal fluid in fasting state; FeSSIF: simulated intestinal fluid in fed state; SGF: simulated gastric fluid in fasting state; the preparation method or process can refer to WO2022063229A1.

[0158] Results: On the one hand, the sample morphology of the free base was improved after the hippurate was prepared; on the other hand, the hippurate sample obtained in Example 1 had good solubility in four different media, and had significantly better solubility in water and FeSSIF (pH 5.0) media than the free base.

[0159] Test Example 2: Hygroscopicity test

[0160] (1) Hygroscopicity test of the sample of Example 1

[0161] Take the sample of Example 1 and perform DVS test.

[0162] Table 6. Hygroscopicity test results of samples of Example 1

[0163]

[0164] Results: It was found that the hippurate salt and its crystal form I obtained in Example 1 only showed slight hygroscopicity under high humidity conditions, and after the DVS test, the sample was tested by X-ray powder diffraction and the crystal form did not change.

[0165] (2) Hygroscopicity test of the sample of Preparation Example 1

[0166] The sample of Preparation Example 1 was taken and placed in an open state at room temperature / 92.5% RH for 24 hours. The sample was then taken for water titration. The results are shown in the table below.

[0167] Table 7. Hygroscopic weight gain test results of the samples of Preparation Example 1

[0168]

[0169] Results: After testing, the sample of Preparation Example 1 showed obvious moisture absorption and weight gain (weight gain of 3.1%) after being placed for 24 hours, indicating that it is hygroscopic. Moreover, after being placed under high humidity conditions for 24 hours, the sample is still in the form of a gel.

[0170] Test Example 3: Stability Test

[0171] (1) Stability test of samples in Example 1

[0172] The sample of Example 1 was weighed and left open at 25°C / 60% RH and 40°C / 75% RH for 1 week respectively. Then the samples were taken for X-ray powder diffraction and purity test respectively to examine the stability of the samples under different conditions. The results are shown in the table below.

[0173] Table 8. Solid stability test results of samples in Example 1

[0174]

[0175] Results: The hippurate salt and its crystal form I of Example 1 were placed under different conditions for one week, and the crystal form remained unchanged, and the purity did not change significantly.

[0176] Table 8.1. XRPD diffraction peak data of the sample of Example 1 after stability test at 25°C / 60%RH

[0177]

[0178] (2) Stability test of sample preparation

[0179] The sample of Preparation Example 1 was taken and placed in an open state under the conditions of 40° C. / 75% RH, and then the sample was taken for testing.

[0180] Results: After testing, the purity of the sample of Preparation Example 1 decreased after being placed for 7 days. Moreover, after being placed under the condition of high humidity for 7 days, the sample was still in the form of a gel.

[0181] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to one skilled in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Claims

1. A compound, which is a hippurate salt of a compound represented by formula (A), 2. The compound according to claim 1, in, The compound is a compound represented by formula (B):

3. The compound according to claim 1 or 2, in, The compound is a compound in solid form.

4. A compound according to any one of claims 1 to 3, in, The solid compound, using the KBr pellet method, has an infrared spectrum including characteristic peaks at the following positions (±4 cm -1 ): 3281, 2939, 1652, 1524, 1373, 1305.

5. A compound according to any one of claims 1 to 3, in, The compound is a compound in crystalline form.

6. A compound according to any one of claims 1 to 5, in, The compound is in crystalline form I, and the X-ray powder diffraction pattern expressed in 2θ angle using Cu-Kα radiation comprises the following characteristic diffraction peaks (°): 6.1±0.3°, 16.0±0.3°, 18.4±0.3°, 20.1±0.3°, 21.7±0.3°; Alternatively, an X-ray powder diffraction pattern at an angle of 2θ comprises the following characteristic diffraction peaks (°): 6.1±0.3°, 13.3±0.3°, 16.0±0.3°, 18.4±0.3°, 20.1±0.3°, 21.7±0.3°; Alternatively, the X-ray powder diffraction pattern at an angle of 2θ comprises the following characteristic diffraction peaks (°): 6.1±0.3°, 8.1±0.3°, 13.3±0.3°, 16.0±0.3°, 18.4±0.3°, 20.1±0.3°, 21.7±0.3°.

7. A crystalline composition comprising the compound of any one of claims 1 to 6.

8. A pharmaceutical composition comprising the compound of any one of claims 1 to 6 or the crystalline composition of claim 7; optionally, further comprising a pharmaceutically acceptable carrier.

9. Use of the compound according to any one of claims 1 to 6, the crystalline composition according to claim 7 or the pharmaceutical composition according to claim 8 in the preparation of a drug; Preferably, the drug is a drug for the prevention and / or treatment of a cell proliferative disease; preferably, the cell proliferative disease is a tumor or cancer; further preferably, the tumor or cancer is a blood tumor or a solid tumor; further preferably, a malignant blood tumor or an advanced solid tumor; further preferably, a relapsed or refractory blood tumor or an advanced malignant solid tumor; or, preferably, The medicine is a medicine for preventing and / or treating a disease at least partially mediated by PRMT5.

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