Crystalline form of bipyrimidine compounds, method of preparation thereof, and use thereof

Abstract

The present invention relates to the crystalline form of bipyrimidine compounds, as well as methods for preparing the same and their use. In particular, the present invention relates to crystalline form I of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine, wherein the X-ray powder diffraction pattern of crystalline form I includes characteristic peaks at diffraction angles (2θ) of 6.378±0.2°, 9.521±0.2°, 15.721±0.2°, 16.379±0.2°, 16.981±0.2°, 17.560±0.2°, 19.080±0.2° and 22.200±0.2°. Crystal form I possesses high stability, high solubility, high plasma concentration, high bioavailability, and excellent pharmacological activity, making it suitable for pharmaceutical preparations.

Description

[Technical Field]

[0001] This invention relates to the crystalline form of bipyrimidine compounds, methods for preparing them, and their uses. [Background technology]

[0002] The duubiquitinase (DUB) family is involved in ubiquitination as a dynamic and reversible process. Humans have approximately 100 DUBs, which can be classified into the cysteine ​​protease family and the metalloproteinase family. Of these, the cysteine ​​protease family mainly includes ubiquitin-specific proteases (USPs), ubiquitin carboxy-terminal hydrolases (UCHs), Machado-Josephin domain proteases (MJDs), MINDY proteases (MINDYs), and ovarian tumor domain proteases (OTUs). The USP family represents the largest number of known duubiquitinases, with over 50 types encoded by human genes. These play a role in various physiological functions, including the cell cycle, signal transduction, DNA damage repair, chromosomal translocation, and gene replication, by regulating protease substrates.

[0003] USP1 belongs to the USP subfamily of DBU and does not exhibit significant activity on its own. However, by binding to UAF1 and forming a heterodimer complex (USP1 / UAF1), it can achieve full enzymatic activity and modulate cellular targets in multiple cancer-related pathways. For example, the USP1 / UAF1 complex can deubiquitinate proliferating cell nuclear antigen (PCNA), which is monoubiquitinated by the major protein during translesion synthesis (TLS), thereby preventing excessive repair. It can also deubiquitinate Fanconi anemia complement D2 (FANCD2), which is monoubiquitinated by the major protein in the Fanconi anemia (FA) pathway, preventing improper TLS repair in cells and thereby ensuring genomic stability. These two DNA damage response (DDR) pathways are important pathways for repairing DNA damage induced by DNA crosslinking agents such as cisplatin, mitomycin, and ultraviolet light. USP1 can also interact with ID proteins and other proteins to stabilize their expression in cells through deubiquitination. For example, in osteosarcoma cells, USP1 can deubiquitinate ID1, ID2, and ID3, promoting cell proliferation, and inhibition of USP1 can increase the sensitivity of osteosarcoma cells to chemotherapy. In addition, there are studies demonstrating a close association between USP1 and the development of resistance to various drugs used for tumor treatment. For example, in cisplatin-resistant non-small cell lung cancer (NSCLC) cells, USP1 is expressed at high levels, and knockdown of USP1 can significantly increase cell sensitivity to cisplatin. In breast cancer cells, USP1 is highly expressed, promoting the proliferation of breast cancer cells and is closely associated with a poor prognosis of breast cancer.As reported in the literature (J. Med. Chem. 2014, 57, pp. 8099-8110, "Synthesis and Structure-Activity Relationship Studies of N-Benzyl-2-phenylpyrimidin-4-amine Derivatives as Potent USP1 / UAF1 Deubiquitinase Inhibitors with Anticancer Activity against Nonsmall Cell Lung Cancer"), compounds such as the USP1 inhibitor ML323 can be used for nonsmall cell lung cancer. As reported in the literature (Cui SZ, Lei ZY, Guan TP et al., "Targeting USP1-dependent KDM4A protein stability as a potential prostate cancer therapy." Cancer Sci. 2020;00:1-15), USP1 inhibitors are used as a potential therapeutic agent for prostate cancer. In summary, USP1 is expected to become a hot target for treating various cancers and other diseases.

[0004] Chinese Patent No. 202310473956.2 discloses a series of USP1 inhibitors, in particular compound 104 having the following structure: 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine.

[0005] [ka]

[0006] However, no prior art has reported on the crystalline form of this compound. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Chinese Patent No. 202310473956.2 [Non-patent literature]

[0008] [Non-Patent Document 1] J. Med. Chem. 2014, 57, pp. 8099-8110, Synthesis and Structure-Activity Relationship Studies of N-Benzyl-2-phenylpyrimidin-4-amine Derivatives as Potent USP1 / UAF1 Deubiquitinase Inhibitors with Anticancer Activity against Nonsmall Cell Lung Cancer [Non-Patent Document 2] Cui SZ, Lei ZY, Guan TP, et al. Targeting USP1-dependent KDM4A protein stability as a potential prostate cancer therapy. Cancer Sci. 2020;00:1~15 [Overview of the project] [Problems that the invention aims to solve]

[0009] The technical problem to be solved by the present invention is to provide the crystalline form of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octane-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine, a method for preparing the same, and its use. The inventors have conducted extensive experimental studies and discovered a method for preparing the crystalline form of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octane-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine. X-ray powder diffraction analysis of the obtained crystalline form revealed a crystalline form I with good stability. [Means for solving the problem]

[0010] The present invention solves the above technical problems through the following technical solutions.

[0011] The present invention provides a crystalline form I of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine, characterized in that its X-ray powder diffraction pattern includes characteristic peaks at diffraction angles (2θ) of 6.378±0.2°, 9.521±0.2°, 15.721±0.2°, 16.379±0.2°, 16.981±0.2°, 17.560±0.2°, 19.080±0.2° and 22.200±0.2°.

[0012] In some embodiments, the X-ray powder diffraction pattern of crystalline form I further includes at least one characteristic peak at diffraction angles (2θ) of 11.938±0.2°, 23.761±0.2°, 25.101±0.2°, and 28.700±0.2°.

[0013] In some embodiments, the X-ray powder diffraction pattern of crystalline form I includes characteristic peaks at diffraction angles (2θ) of 6.378 ± 0.2°, 9.521 ± 0.2°, 11.938 ± 0.2°, 15.721 ± 0.2°, 16.379 ± 0.2°, 16.981 ± 0.2°, 17.560 ± 0.2°, 19.080 ± 0.2°, 22.200 ± 0.2°, 23.761 ± 0.2°, 25.101 ± 0.2° and 28.700 ± 0.2°.

[0014] In some embodiments, the X-ray powder diffraction pattern of crystalline form I further includes at least one characteristic peak at diffraction angles (2θ) of 10.239 ± 0.2°, 12.400 ± 0.2°, 14.882 ± 0.2°, 20.420 ± 0.2° and 25.980 ± 0.2°.

[0015] In some embodiments, the X-ray powder diffraction pattern of crystalline form I includes characteristic peaks at diffraction angles (2θ) of 6.378 ± 0.2°, 9.521 ± 0.2°, 10.239 ± 0.2°, 11.938 ± 0.2°, 12.400 ± 0.2°, 14.882 ± 0.2°, 15.721 ± 0.2°, 16.379 ± 0.2°, 16.981 ± 0.2°, 17.560 ± 0.2°, 19.080 ± 0.2°, 20.420 ± 0.2°, 22.200 ± 0.2°, 23.761 ± 0.2°, 25.101 ± 0.2°, 25.980 ± 0.2° and 28.700 ± 0.2°.

[0016] In some embodiments, the X-ray powder diffraction pattern of crystalline form I is substantially as shown in FIG. 1.

[0017] In some embodiments, the DSC spectrum of crystalline form I shows one exothermic peak at 219.25 ± 2 °C.

[0018] In some embodiments, the TGA spectrum of crystalline form I shows a mass loss of 0.10% at 20 °C to 120 °C and a mass loss of 0.37% at 120 °C to 250 °C.

[0019] The present invention also relates to a method for preparing crystalline form I, comprising the following steps: A step of mixing 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine with an organic solvent, and a step of collecting the solid after solid-liquid separation of the resulting suspension. Includes, The organic solvent is one or more selected from the group consisting of petroleum ethers, C4-C6 ethers, and C2-C6 nitriles. Provide a method.

[0020] In some embodiments, the method involves the following steps: The process involves grinding 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine in an organic solvent at 0 to 30°C for 10 minutes to 2 hours, preferably at 25°C for 30 minutes; collecting the solid after solid-liquid separation of the resulting suspension; and optionally drying the solid. Includes, The organic solvent is one or more selected from the group consisting of petroleum ether, methyl tert-butyl ether, tetrahydrofuran, and acetonitrile, preferably a mixture of petroleum ether and methyl tert-butyl ether. Advantageously, the ratio (w / v) of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine to the organic solvent is 1:5 to 1:1, preferably 1:3.

[0021] The present invention also relates to a method for preparing crystalline form I, comprising the following steps: A step of mixing 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine with a solvent, and a step of collecting the solid after solid-liquid separation of the resulting suspension. This provides a method that includes this.

[0022] In some embodiments, the method includes one of the following methods: Method (I) consists of the following steps: A step of grinding 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine in a solvent, and a step of collecting the solid after solid-liquid separation of the resulting suspension. Includes, The solvent is one or more selected from the group consisting of water, petroleum ether, C1-C6 alcohols, C4-C8 alkanes, C4-C6 ethers, C3-C6 ketones, C3-C8 esters, toluene, and C2-C6 nitriles. Method (II) involves the following steps: The process involves mixing 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine with a good solvent to dissolve it, then mixing the resulting solution with an anti-solvent, and precipitating the crystals to obtain crystalline form I. Includes, A good solvent is one or more selected from the group consisting of C2-C6 nitriles, methanol, benzyl alcohol, C3-C6 ketones, N-methylpyrrolidone, ethyl acetate, dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, and tetrahydrofuran. The poor solvent is one or more selected from the group consisting of petroleum ethers, C4-C6 ethers, and C2-C6 alcohols. Method (III) involves the following steps: The steps are: mixing 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine and a solvent; cooling the resulting mixture; and precipitating the crystals to obtain crystalline form I. Includes, The solvent is one or more selected from the group consisting of C2-C6 nitriles, C1-C3 alcohols, benzyl alcohol, C3-C6 ketones, N-methylpyrrolidone, C3-C8 esters, dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, methyltetrahydrofuran, and 1,4-dioxane.

[0023] In some embodiments, method (I) includes the following steps: The process involves grinding 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine in a solvent at 0-50°C for 10 minutes to 7 days, preferably at 25-30°C for 30 minutes to 4 days; collecting the solid after solid-liquid separation of the resulting suspension; and optionally drying the solid. Includes, The solvent is one or more selected from the group consisting of water, methanol, n-heptane, ethyl acetate, ethylene glycol, isopropyl acetate, methyl isobutyl ketone, toluene, petroleum ether, methyl tert-butyl ether, tetrahydrofuran, and acetonitrile. Preferably, the solvent is a mixture of petroleum ether and methyl tert-butyl ether, a mixture of water and ethanol, a mixture of n-heptane and ethyl acetate, a mixture of ethylene glycol and methanol, isopropyl acetate, methyl isobutyl ketone, and toluene. More preferably, the volume ratio of petroleum ether to methyl tert-butyl ether is 1:10 to 10:1, preferably 1:1 to 10:1; the volume ratio of water to ethanol is 5:1 to 30:1, preferably 10:1 to 20:1; the volume ratio of n-heptane to ethyl acetate is 5:1 to 30:1, preferably 10:1 to 20:1; and the volume ratio of ethylene glycol to methanol is 5:1 to 30:1, preferably 10:1 to 20:1. Advantageously, the ratio (w / v) of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine to the solvent is 1g:1mL to 1g:20mL, preferably 1g:3mL to 1g:10mL.

[0024] In some embodiments, method (II) includes the following steps: The process involves dissolving 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine by mixing it with a good solvent at 5 to 45°C, preferably at room temperature; then mixing the resulting solution with a poor solvent; and precipitating the crystals to obtain crystalline form I. Includes, A good solvent is one or more selected from the group consisting of methanol, acetone, ethyl acetate, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran, acetonitrile, and benzyl alcohol. The poor solvent is one or more selected from the group consisting of methyl tert-butyl ether, isopropanol, 1-butanol, and ethylene glycol. Advantageously, the volume of the poor solvent is 2 to 20 times, preferably 3 to 10 times, the volume of the good solvent. More favorably, the ratio (w / v) of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine to the poor solvent is 50 mg:1 mL to 1 mg:1 mL, preferably 50 mg:1 mL to 5 mg:1 mL.

[0025] In some embodiments, method (II) includes the following steps: The process involves: mixing 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine with a good solvent at 5 to 45°C, preferably at room temperature, to obtain a saturated or near-saturated solution; optionally filtering the solution; then mixing the solution with a poor solvent, preferably under stirring; and precipitating the crystals to obtain crystalline form I. Includes.

[0026] In some embodiments, method (III) includes the following steps: The steps are: mixing 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine and a solvent; cooling the resulting mixture; and precipitating the crystals to obtain crystalline form I. Includes, The solvent is a good solvent, and preferably, the solvent is one or more selected from the group consisting of ethanol, methyl isobutyl ketone, isopropyl acetate, methyl tetrahydrofuran, and 1,4-dioxane.

[0027] Preferably, method (III) consists of the following steps: The process involves mixing and dissolving 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine with a good solvent; optionally filtering the resulting mixture; first cooling the filtrate until a solid precipitate forms; then further cooling the filtrate; and precipitating the crystals to obtain crystalline form I. Includes.

[0028] Preferably, method (III) consists of the following steps: The process involves: dissolving 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine by mixing it with a good solvent at 25-60°C, preferably 30-50°C; optionally filtering the resulting mixture; first cooling the filtrate to 5-35°C, preferably room temperature; then allowing the mixture to stand at -5-5°C, preferably 4°C; and precipitating the crystals to obtain crystalline form I. Includes, More preferably, the ratio (w / v) of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine to the solvent is 100 mg:1 mL to 10 mg:1 mL, preferably 80 mg:1 mL to 30 mg:1 mL.

[0029] The present invention also provides a pharmaceutical composition comprising crystalline form I and one or more pharmaceutically acceptable carriers.

[0030] The present invention relates to the use of crystalline form I or a pharmaceutical composition containing it in the preparation of a pharmaceutical for treating or preventing a disease or condition associated with the inhibition of ubiquitin-specific protease 1 (USP1).

[0031] The present invention also relates to the use of crystalline form I or a pharmaceutical composition containing it in the preparation of a pharmaceutical for treating or preventing cancer.

[0032] The present invention also relates to crystalline form I or a pharmaceutical composition containing the same for use in the treatment or prevention of diseases or conditions associated with the inhibition of ubiquitin-specific protease 1 (USP1).

[0033] The present invention also relates to crystalline form I or a pharmaceutical composition containing the same for use in the treatment or prevention of cancer.

[0034] The present invention also relates to a method for treating or preventing a disease or condition associated with the inhibition of ubiquitin-specific protease 1 (USP1), comprising the step of administering an effective amount of crystalline form I or a pharmaceutical composition containing the same to a patient in need.

[0035] The present invention also relates to a method for treating or preventing cancer, comprising the step of administering an effective amount of crystalline form I or a pharmaceutical composition containing the same to a patient in need.

[0036] In some embodiments, cancer is selected from the group consisting of lung cancer, non-small cell lung cancer (NSCLC), colon cancer, bladder cancer, osteosarcoma, ovarian cancer, skin cancer, and breast cancer.

[0037] The term "pharmaceutically acceptable" as used in this disclosure means that the material is generally safe, free from biological toxicity and other undesirable toxicity, and useful for preparing pharmaceutical compositions that are acceptable for veterinary and human pharmaceutical use.

[0038] As used in this disclosure, the term “carrier” refers to diluents, adjuvants, or excipients administered with a compound, and includes, but is not limited to, fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, matrices, etc. The compositions of the present invention may be prepared by any method known in the art to provide rapid, sustained, or slow release of the active ingredient after administration to a patient. [Brief explanation of the drawing]

[0039] [Figure 1] This is the X-ray powder diffraction (XRPD) pattern of crystal morphology I. [Figure 2] This is the differential scanning calorimetry (DSC) spectrum of crystal morphology I. [Figure 3] This is the thermogravimetric analysis (TGA) spectrum of crystal morphology I. [Figure 4] This is the PLM spectrum (10X10X) of crystal morphology I. [Figure 5] This is the PLM spectrum (10X40X) of crystal morphology I. [Figure 6] This is the result of DVS for crystal morphology I. [Figure 7] These are the XRPD patterns of crystal morphology I before and after DVS. In this figure, from top to bottom, are the XRPD patterns after DVS and before DVS. [Modes for carrying out the invention]

[0040] The present invention will be described in more detail through the following embodiments. The embodiments of the present invention are intended merely to describe the technical solutions of the present invention and should not be considered as limiting the spirit and scope of the invention.

[0041] The purity analysis method is as follows: A Kinetex EVO C18 (50 × 4.6 mm, 5 μm, 100 Å) chromatography column was used, with acetonitrile-water as the mobile phase for gradient elution, a flow rate of 1.5 ml / min, and a detection wavelength of 220 nm.

[0042] The mass spectrometer (MS) is determined by an LC (Agilent 1260 Infinity II) / MS (G6125B single quadrupole) mass spectrometer (manufactured by Agilent) (photodiode array detector).

[0043] The compound's structure was confirmed by hydrogen nuclear magnetic resonance, with the instrument model being WNMR-I-400MHz.

[0044] Preparative liquid chromatography is performed using an Agilent 1260 Infinity II high-performance liquid chromatograph (manufacturer: Agilent). The chromatography column is Daisogel C18 10μm 100A (30mm × 250mm), and the mobile phase is acetonitrile / water.

[0045] Qingdao Haiyang Chemical's GF254 silica gel plates are used for thin-layer silica gel chromatography (TLC). The silica gel plates used for TLC have dimensions of 0.20 mm to 0.25 mm, while the silica gel plates used for product purification have dimensions of 0.5 mm.

[0046] Silica gel of 100-200 mesh, 200-300 mesh, or 300-400 mesh from Qingdao Haiyang Chemical is used as a support for gel column chromatography.

[0047] The known starting materials of the present invention can be prepared by methods known in the art, or can be purchased from Wanghua Mall, Beijing Ouhe Technology, Sigma, J&K Scientific, Yishiming, Shanghai Shuya Chemical, Shanghai Innochem Science & Technology, Energy Chemical, Shanghai Bide Pharmatech, etc.

[0048] Unless otherwise specified in the examples, the reactions are carried out under a nitrogen atmosphere.

[0049] A nitrogen atmosphere means that the reaction flask is equipped with a nitrogen balloon (approximately 1 L).

[0050] Unless otherwise specified in the examples, "solution" means an aqueous solution.

[0051] The chemical reactions described in this invention are generally carried out under atmospheric pressure. Unless otherwise specified in the examples, the reaction temperature is room temperature, i.e., 20°C to 30°C.

[0052] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those well known to those skilled in the art. In addition, any methods and materials similar to or equivalent to those described herein can be applied to the methods of the present invention.

[0053] Unless otherwise specified, the mixing ratio of different solvents is given as a volume ratio.

[0054] X-ray powder diffraction (XRPD) The instrument used is a Shimadzu XRD-6000, and the sample is scanned according to the following parameters. The radiation source is a Cu~Kα target (1.54056 Å). The minimum operating voltage and current for the phototube are 40kV and 30mA, respectively. The 2-theta value of the sample scanning range is 2° to 40°. The scanning speed is 4° / min.

[0055] Thermogravimetric analysis (TGA) The instrument used is a PerkinElmer Pyris1 TGA. The analysis is performed at a heating rate of 10°C / min from 20°C to 350°C, and held at 350°C for 1 minute.

[0056] Differential Scanning Calorimetry (DSC) The instrument used is a METTLER DSC3. Differential scanning calorimetry is performed from 30°C to 300°C, with a holding time of 1 minute at 300°C. The heating rate is 10°C / min.

[0057] Polarizing microscope (PLM) The PLM equipment is an XPV-203E manufactured by Shanghai Changfang Optical Instrument Co., Ltd. The sample is observed using a 10x eyepiece and a 10x or 40x objective lens. Images are recorded using a camera-computer system.

[0058] Dynamic vapor sorption (DVS) The DVS system is an SMS DVS Intrinsic. The characteristics of moisture adsorption / desorption are tested in an aqueous environment at 25°C under a cycle of relative humidity (RH) from 0% to 90% to 0%.

[0059] HPLC HPLC analysis for solubility and stability is performed using a Phenomenex Luna C18(2), 3 μm, 2 × 50 mm chromatography column. [Examples]

[0060] (Example 1) Synthesis of 4'-Cyclopropyl-5,6'-Dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine 1.1 Synthesis of General Intermediate A2-7 ((4-Cyclopropyl-6-methoxypyrimidine-5-yl)boronic Acid)

[0061] [ka]

[0062] Step 1: Synthesis of compound A2-3 (6-cyclopropylpyrimidine-4-ol) Compound A2-1 (200.0 g, 1.4 mol, 1.00 equivalent) and compound A2-2 (292.0 g, 2.81 mol, 2.00 equivalent) were added to methanol (1.2 L), and a solution of sodium methoxide in methanol (5.4 M, 1.3 L, 5.00 equivalent) was added at 0°C. After the addition was complete, the reaction was heated to 20°C and stirred. Glacial acetic acid was added at 0°C to adjust the pH to 7-8. The resulting mixture was concentrated under vacuum to obtain the crude product, which was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-1 / 1) to obtain compound A2-3 (90.3 g). LC-MS: m / z = 137.1 (M + H) + .

[0063] Step 2: Synthesis of compound A2-4 (4-chloro-6-cyclopropylpyrimidine) Compound A2-3 (40.0 g, 293.0 mmol, 1.00 equivalent) was added to phosphorus oxychloride (180.0 mL), and the reaction mixture was heated to 60°C and stirred. The reaction solution was concentrated under vacuum to obtain the crude product. The crude product was dissolved in ethyl acetate (400.0 mL) and water (400.0 mL) and extracted with ethyl acetate (400.0 mL). The organic layer was dried and filtered to obtain the filtrate. The filtrate was concentrated under vacuum to obtain the crude product, which was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 20 / 1~1 / 1) to obtain compound A2-4 (11.0 g). LC-MS: m / z = 155.0 (M+H) + .

[0064] Step 3: Synthesis of compound A2-5 (5-bromo-4-chloro-6-cyclopropylpyrimidine) Compound A2-4 (11.0 g, 71.1 mmol, 1.00 equivalent) was dissolved in methanol (150.0 mL), and bromine (34.1 g, 213.0 mmol, 3.00 equivalent) was added at -60°C. The reaction was heated to 20°C and stirred. Then, saturated sodium bicarbonate solution (200.0 mL) and water (100.0 mL) were added at 0°C, and the resulting mixture was extracted three times with dichloromethane (200.0 mL). The organic layer was washed with water, dried, and filtered to obtain the filtrate. The filtrate was concentrated under vacuum to obtain the crude product, which was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0 to 3 / 1) to obtain compound A2-5 (7.8 g). 1 H NMR (400 MHz, CDCl3) δ 8.61 (s, 1H), 2.56 - 2.62 (m, 1H), 1.23 - 1.26 (m, 2H), 1.16 - 1.21 (m, 2H); LC-MS: m / z = 232.9 (M+H) + .

[0065] Step 4: Synthesis of compound A2-6 (5-bromo-4-cyclopropyl-6-methoxypyrimidine) Compound A2-5 (7.8 g, 33.4 mmol, 1.00 equivalent) was dissolved in methanol (240.0 mL), and sodium methoxide (18.0 g, 100.0 mmol, 3.00 equivalent) was added at 0°C. The reaction was heated to 30°C and stirred. The reaction solution was concentrated under vacuum to obtain the crude product, which was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to obtain compound A2-6 (7.3 g). LC-MS: m / z = 228.9 (M + H) + .

[0066] Step 5: Synthesis of compound A2-7 ((4-cyclopropyl-6-methoxypyrimidine-5-yl)boronic acid) Compound A2-6 (10.3 g, 44.9 mmol, 1.00 equivalent) and triisopropyl borate (11.8 g, 62.9 mmol, 1.40 equivalent) were dissolved in tetrahydrofuran (30.0 mL) and toluene (90.0 mL), and n-butyllithium (2.5 M, 25.1 mL, 1.40 equivalent) was added dropwise at -70°C. The reaction was stirred at -70°C. Next, 50.0 mL of 1N hydrochloric acid solution was added dropwise at -70°C. The reaction was heated to 20°C and stirred. Then, saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8, and the resulting mixture was extracted three times with ethyl acetate (100.0 mL). The organic layer was washed with water, dried, filtered, and the filtrate was obtained, which was concentrated under vacuum to obtain compound A2-7 (6.9 g). 1 H NMR (400 MHz, CDCl3) δ 8.52 (s, 1H), 8.45 (s, 2H), 3.84 (s, 3H), 1.88 - 1.92 (m, 2H), 0.93 - 1.01 (m, 4H).

[0067] 1.2 General intermediate BB2 (2-chloro-5-methoxypyrimidine-4-amine) Intermediate BB2 was purchased from Leyan Reagents.

[0068] [ka]

[0069] 1.3 Synthesis of the general intermediate C35 ((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl-4-methylbenzenesulfonate)

[0070] [ka]

[0071] Step 1: Synthesis of compound C35-2 (1-methyl-4-(trifluoromethyl)-1H-imidazole) Compound 4-(trifluoromethyl)-1H-imidazole (Compound C35-1, 26 g, 0.19 mol) and potassium carbonate (105.5 g, 0.76 mol, 4.0 equivalents) were added to acetonitrile (390 mL). Iodomethane (32.6 g, 0.23 mol, 1.2 equivalents) was added dropwise at 0 °C, and the reaction mixture was stirred at room temperature. Subsequently, the reaction solution was concentrated under vacuum to obtain a crude product, which was extracted three times with ethyl acetate (200 mL). The organic layer was washed with water, dried, filtered to obtain a filtrate, which was concentrated under vacuum to obtain Compound C35-2 (26 g, crude). LC-MS: m / z = 151.0 (M+H) + .

[0072] Step 2: Synthesis of Compound C35-3 (methyl 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)bicyclo[2.2.2]octane-1-carboxylate) Compound C35-2 (14.75 g, 98.3 mmol) and monomethyl hydrogen bicyclo[2.2.2]octane-1,4-dicarboxylate (25 g, 117.9 mmol, 1.2 equivalents) were dissolved in dichloromethane (295 mL) and water (295 mL), and subsequently silver nitrate (6.0 g, 35.4 mmol, 0.3 equivalents) and ammonium persulfate (44.9 g, 78.7 mmol, 2.0 equivalents) were added. The reaction mixture was stirred at 25 °C, filtered through diatomaceous earth, and extracted three times with dichloromethane (200 mL). The organic layer was washed with water, dried, filtered to obtain a filtrate, which was concentrated under vacuum to obtain Compound C35-3 (35.2 g, crude) as a yellow oil. 1 H NMR (400 MHz, CDCl3) 7.09 - 7.08 (m, 1H), 3.77 (s, 3H), 3.67 (s, 3H), 2.07 - 2.03 (m, 6H), 1.93 - 1.89 (m, 6H); LC-MS: m / z = 317.2 (M+H) + .

[0073] Step 3: Synthesis of compound C35-4 ((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methanol) Compound C35-3 (35.2 g, 111.4 mmol) was dissolved in tetrahydrofuran (180.0 mL), and lithium aluminum hydride (2.5 M, 111.4 mL, 278.5 mmol, 2.5 equivalents) was added at 0°C. The reaction mixture was heated to 20°C and stirred. Then, ice water (50.0 mL) was added at 0°C, and the resulting mixture was extracted three times with ethyl acetate (500.0 mL). The organic layer was dried and filtered to obtain the filtrate, which was concentrated under vacuum to obtain compound C35-4 (25.1 g, crude) as a yellow oily substance. LC-MS: m / z = 289.1 (M + H) + .

[0074] Step 4: Synthesis of compound C35 Compound C35-4 (25.1 g, 87.2 mmol), p-toluenesulfonyl chloride (33.2 g, 174.4 mmol, 2.0 equivalents), and 4-dimethylaminopyridine (32.0 g, 261.6 mmol, 3.0 equivalents) were sequentially added to dichloromethane (251 mL), and the reaction mixture was stirred at 25°C. The reaction solution was washed with water, dried, and filtered to obtain the filtrate. The filtrate was concentrated under vacuum to obtain the crude product, which was purified by column chromatography (eluent: petroleum ether / dichloromethane = 1 / 0 to 0 / 1) to obtain intermediate C35 (10.0 g) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 8.2 Hz, 2H), 7.67 (s, 1H), 7.52 (d, J = 8.0 Hz, 2H), 3.77 (s, 3H), 3.72 (s, 2H), 2.46 (s, 3H), 1.95 - 1.86 (m, 6H), 1.49 - 1.37 (m, 6H); LC-MS: m / z = 443.3 (M+H) + .

[0075] 1.4 Synthesis of the intermediate BB2C35(2-chloro-5-methoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)pyrimidine-4-amine)

[0076] [ka]

[0077] Compound BB2 (1.0 g, 6.29 mmol, 1.00 equivalent), compound C35 (2.78 g, 6.29 mmol, 1.0 equivalent), and cesium carbonate (10.2 g, 31.45 mmol, 5.0 equivalents) were sequentially added to N,N-dimethylformamide (20.0 mL), and the reaction mixture was stirred at 130°C. The reaction solution was cooled to 25°C. Dichloromethane (100.0 mL) was added, and the reaction solution was filtered to obtain the filtrate. The filtrate was washed with water, dried, and concentrated under vacuum to obtain the crude product, which was purified by column chromatography (eluent: petroleum ether / dichloromethane = 1 / 0~0 / 1) to obtain the intermediate BB2C35 (1.03 g, 2.41 mmol, yield 38.3%) as a white solid. LC-MS: m / z = 430.1 (M+H) + .

[0078] 1.5 Synthesis of 4'-Cyclopropyl-5,6'-Dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-Bipyrimidine]-4-amine

[0079] [ka]

[0080] Compound BB2C35 (85.8 g, 0.2 mol), compound A2-7 (48.5 g, 0.25 mol, 1.25 equivalents), ligand XPhos (37.4 g, 0.08 mol, 0.4 equivalents), potassium phosphate (169.8 g, 0.8 mol, 4 equivalents), and catalyst XPhos-Pd-G2 (31.5 g, 0.04 mol, 0.2 equivalents) were sequentially added to 1,4-dioxane (1026 mL) / water (171.6 mL). The reaction mixture was heated to 85°C and stirred under a nitrogen atmosphere. The mixture was concentrated under reduced pressure, cooled to 25°C, and dissolved in water. The resulting solution was then extracted with ethyl acetate (1026 mL), dried, and concentrated under reduced pressure to obtain approximately 200 g of the crude labeled compound.

[0081] Here, XPhos is 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, and XPhos-Pd-G2 is chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II).

[0082] (Example 2) Preparation of crystalline form I of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine Approximately 200 g of the crude product of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine obtained in Example 1 was dissolved in 1 L (5V) ethyl acetate. Silica gel was then added, and the resulting mixture was concentrated under vacuum until it became a powder. The powder was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 100:5) to obtain approximately 91 g of a pale yellow solid. The solid was then pulverized in 275 ml of solvent (petroleum ether:methyl tert-butyl ether = 10:1) at 25°C for 30 minutes. The resulting mixture was filtered and dried under vacuum at 50°C to obtain 85.0 g of a white powder solid in yield 78% and purity 99.2%. LC-MS: m / z = 544.3 (M+H) + ; 1 H NMR (400 MHz, DMSO-d6) δ 8.62 (s, 1H), 7.90 (s, 1H), 7.65 (d, J = 1.5 Hz, 1H), 6.99 (t, J = 6.5 Hz, 1H), 3.93 (s, 3H), 3.84 (s, 3H), 3.77 (s, 3H), 3.21 (d, J = 6.5 Hz, 2H), 1.90 (m, 6H), 1.74 (m, 1H), 1.48 (m, 6H), 1.06 - 1.01 (m, 2H), 0.92 - 0.85 (m, 2H).

[0083] After X-ray powder diffraction analysis, the crystal morphology was defined as crystal morphology I. The X-ray powder diffraction data is shown in Table 1, and the X-ray powder diffraction pattern is shown in Figure 1.

[0084] The DSC spectrum of crystal morphology I is shown in Figure 2, which exhibits a characteristic exothermic peak at 219.25±2°C.

[0085] The TGA spectrum of crystal morphology I is shown in Figure 3, which exhibits a mass loss of 0.10% from 20°C to 120°C and a mass loss of 0.37% from 120°C to 250°C.

[0086] The PLM results for crystal morphology I are shown in Figures 4 and 5.

[0087] Figure 6 shows the DVS results for crystal morphology I, which shows a water absorption rate of 0.02% from 0%RH to 95%RH, indicating that crystal morphology I is non-hygroscopic. Figure 7 shows the XRPD pattern of the crystal morphology after DVS, which indicates that the crystal morphology has not changed after DVS.

[0088] [Table 1]

[0089] (Examples 3-10) Preparation of crystal morphology I (poor solvent method) Approximately 50 mg of crystalline form I was weighed and dissolved at room temperature in an appropriate volume of the good solvent shown in Table 2 to obtain a saturated or near-saturated solution. The solution was filtered, and the poor solvent shown in Table 2 was slowly added under magnetic stirring in an amount 3 to 10 times the volume of the good solvent. The resulting mixture was stirred until a sufficient amount of solid precipitated, and then the solid was collected by centrifugation. The obtained solid was dried under reduced pressure (40°C).

[0090] [Table 2]

[0091] X-ray powder diffraction analysis showed that all crystal forms obtained in Examples 3-10 were crystal form I.

[0092] (Examples 11-16) Preparation of crystalline form I (suspension-grinding method) Approximately 50 mg of crystalline form I was weighed, and an appropriate volume of the solvent shown in Table 3 was added. The resulting mixture was magnetically stirred (400 rpm) at 50°C for 4 days, and then centrifuged to collect the solid.

[0093] [Table 3]

[0094] X-ray powder diffraction analysis showed that all crystal forms obtained in Examples 11-16 were crystal form I.

[0095] (Examples 17-21) Preparation of crystal morphology I (cooling method) Approximately 50 mg of crystalline form I was weighed and dissolved at 50°C in an appropriate volume of a good solvent shown in Table 4. The resulting solution was filtered and cooled to room temperature until the solid precipitated, then placed in a refrigerator at 4°C. Once sufficient solid had precipitated, the mixture was filtered to collect the solid, which was then dried under reduced pressure (40°C).

[0096] [Table 4]

[0097] X-ray powder diffraction analysis showed that all crystal forms obtained in Examples 17-21 were crystal form I.

[0098] (Example 22) Research on the grinding of crystal morphology I Approximately 50 mg of crystalline form I was manually ground in a mortar using the conditions shown in Table 5, and the crystalline form of the ground solid was examined by performing XRPD.

[0099] [Table 5]

[0100] The results showed that all crystal forms obtained from the above grinding studies were crystal form I. Crystal form I has good grinding stability.

[0101] (Example 23) Solubility study of crystal morphology I 50 μL or 100 μL of the solvent shown in Table 6 was gradually added to 2 mg of crystalline form I, and the resulting mixture was dissolved by manual shaking, vortexing, or sonication. Addition of the solvent was stopped when the concentration fell below 1 mg / mL. Solubility was calculated based on the sample volume and the total amount of solvent added. The results are shown in Table 6.

[0102] [Table 6]

[0103] The results showed that crystalline form I had good solubility in organic solvents, moderate solubility in low pH solutions (pH 1.2) ranging from 8.45 to 16.9 mg / mL, and poor solubility in high pH buffer solutions (pH 7.4 phosphate buffer) of 0.31 mg / mL after 24 hours at 37°C.

[0104] (Example 24) Stability study of crystal morphology I (1) Stability against forced decomposition 3 mg of crystalline form I was weighed, and a 1 mg / mL storage solution was prepared using 3 mL of 90% acetonitrile. The stability of the solution was studied under oxidative decomposition (decomposition with 3% H2O2 at 40°C for 1.5 hours) and alkaline decomposition (decomposition with 0.1N NaOH at 40°C for 2 hours).

[0105] The results show that crystalline form I exhibits only slight decomposition after incubation in 0.1 NaOH at 40°C for 2 hours, or in 3% H2O2 at 40°C for 1.5 hours, indicating good stability.

[0106] (2) Stability when exposed to temperature and humidity 1-3 mg of crystalline form I solid powder was weighed and left to stand at 60°C or 60°C / 75%RH for 30 days. Samples were collected for LC studies and analysis.

[0107] The results showed that the composition of the main components of crystalline form I changed by less than 0.2% at 60°C or 60°C / 75%RH, and remained relatively stable over 30 days.

[0108] In addition, an appropriate amount of solid powder in crystalline form 1 was weighed and left to stand for 30 days under stable conditions (40°C / 75%RH, 60°C / 75%RH). The samples were collected for XRPD.

[0109] The results showed that crystal morphology I did not change after being left standing for 30 days at 40°C / 75%RH and 60°C / 75%RH.

[0110] (3) Stability when exposed to light An appropriate amount of crystalline form I solid powder was weighed, placed in a light-stabilized chamber, and irradiated under ICH light intensity conditions (at least greater than 1.2*10^6 Lux*h, illuminance 4500±500 Lux, and intensity 200 w*hr / m). 2 The samples were collected for LC research and analysis.

[0111] The results indicate that the content of the main components of crystal morphology I changed by less than 0.2%, suggesting that it is photostability.

[0112] (Example 25) Pharmacokinetic studies of intragastric administration of suspensions Crystallized form I was used in pharmacokinetic studies in rats. Twelve male SD rats were weighed before administration and housed using standard feeding methods. The animals were housed in standard cages with free access to food and water during the study. The rats were administered the suspension of crystallized form I intragastricly. 40 μL of blood was collected at each sampling point and placed in EDTA-containing anticoagulant vessels. After centrifugation, 20 μL of plasma was collected. All samples were frozen in a -80°C refrigerator immediately after collection.

[0113] The results demonstrate a significant pharmacokinetic advantage, showing that intragastric administration of the crystalline form I suspension resulted in high plasma concentrations, high exposure, and high bioavailability in SD rats.

Claims

1. Crystal morphology I of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine, characterized in that its X-ray powder diffraction pattern includes characteristic peaks at diffraction angles (2θ) of 6.378±0.2°, 9.521±0.2°, 15.721±0.2°, 16.379±0.2°, 16.981±0.2°, 17.560±0.2°, 19.080±0.2° and 22.200±0.2°.

2. The crystal morphology I according to claim 1, characterized in that the X-ray powder diffraction pattern further includes at least one characteristic peak at diffraction angles (2θ) of 11.938±0.2°, 23.761±0.2°, 25.101±0.2°, and 28.700±0.2°.

3. The crystal morphology I according to claim 1 or 2, characterized in that the X-ray powder diffraction pattern includes characteristic peaks at diffraction angles (2θ) of 6.378±0.2°, 9.521±0.2°, 11.938±0.2°, 15.721±0.2°, 16.379±0.2°, 16.981±0.2°, 17.560±0.2°, 19.080±0.2°, 22.200±0.2°, 23.761±0.2°, 25.101±0.2° and 28.700±0.2°.

4. The crystal morphology I according to any one of claims 1 to 3, characterized in that the X-ray powder diffraction pattern further includes at least one characteristic peak at diffraction angles (2θ) of 10.239±0.2°, 12.400±0.2°, 14.882±0.2°, 20.420±0.2°, and 25.980±0.2°.

5. The crystal morphology I according to any one of claims 1 to 4, characterized in that the X-ray powder diffraction pattern includes characteristic peaks at diffraction angles (2θ) of 6.378±0.2°, 9.521±0.2°, 10.239±0.2°, 11.938±0.2°, 12.400±0.2°, 14.882±0.2°, 15.721±0.2°, 16.379±0.2°, 16.981±0.2°, 17.560±0.2°, 19.080±0.2°, 20.420±0.2°, 22.200±0.2°, 23.761±0.2°, 25.101±0.2°, 25.980±0.2° and 28.700±0.2°.

6. Crystal morphology I according to any one of claims 1 to 5, characterized in that the DSC spectrum shows one exothermic peak at 219.25±2℃.

7. Crystal morphology I according to any one of claims 1 to 6, characterized in that the TGA spectrum shows a mass loss of 0.10% at 20°C to 120°C and a mass loss of 0.37% at 120°C to 250°C.

8. A method for preparing the crystalline form I described in any one of claims 1 to 7, comprising the following steps: A step of mixing 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine with an organic solvent, and a step of collecting the solid after solid-liquid separation of the resulting suspension. Includes, The aforementioned organic solvent is petroleum ether, C 4 ~C 6 Ether and C 2 ~C 6 A method comprising one or more selected from the group consisting of nitriles.

9. The following steps: The process involves grinding 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine in an organic solvent at 0 to 30°C for 10 minutes to 2 hours, preferably at 25°C for 30 minutes; collecting the solid after solid-liquid separation of the resulting suspension; and optionally drying the solid. Includes, The organic solvent is one or more selected from the group consisting of petroleum ether, methyl tert-butyl ether, tetrahydrofuran, and acetonitrile, preferably a mixture of petroleum ether and methyl tert-butyl ether. Advantageously, the ratio (w / v) of 4'-cyclopropyl-5,6'-dimethoxy-N-((4-(1-methyl-4-(trifluoromethyl)-1H-imidazole-2-yl)bicyclo[2.2.2]octan-1-yl)methyl)-[2,5'-bipyrimidine]-4-amine to the organic solvent is 1:5 to 1:1, preferably 1:

3. The method according to claim 8.

10. A pharmaceutical composition comprising the crystalline form I described in any one of claims 1 to 7 and one or more pharmaceutically acceptable carriers.

11. Use of the crystalline form I according to any one of claims 1 to 7 or the pharmaceutical composition according to claim 10 in the preparation of a pharmaceutical for treating or preventing a disease or condition associated with the inhibition of ubiquitin-specific protease 1 (USP1).

12. The use of crystalline form I according to any one of claims 1 to 7 or the pharmaceutical composition according to claim 10 in the preparation of a pharmaceutical for treating or preventing cancer, In particular, the aforementioned cancer is selected from the group consisting of lung cancer, non-small cell lung cancer (NSCLC), colon cancer, bladder cancer, osteosarcoma, ovarian cancer, skin cancer, and breast cancer. use.

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