Crystalline forms of CDK inhibitors and pharmaceutically acceptable salts thereof and uses thereof

Abstract

The present invention provides a crystalline form of Compound A or a pharmaceutically acceptable salt thereof, whose structure is represented by the following formula (I): Studies have shown that the free base crystalline form AI, the hydrochloride crystalline form B, and the benzenesulfonate crystalline form C have certain physical and chemical stability. The crystalline form of Compound A or a pharmaceutically acceptable salt thereof can significantly inhibit the growth of various tumor cells, including breast cancer, colon cancer, ovarian cancer, malignant melanoma, liver cancer, lung cancer, and acute myeloid leukemia, and has excellent inhibitory effects against palbociclib-resistant (primary resistance and acquired resistance) breast cancer and liver cancer. Therefore, the crystalline form of Compound A or a pharmaceutically acceptable salt thereof has excellent clinical applicability in the treatment of advanced malignant tumors, providing a new drug option for the treatment of CDK4 / 6 inhibitor-resistant tumors. [Formula 1] JPEG2026502614000043.jpg64119

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to and the benefit of Chinese patent application No. 202310199886.6, filed with the State Intellectual Property Office of China on January 17, 2023, the entire disclosure of which is incorporated herein by reference.

[0002] The present application is in the field of medicine, and specifically relates to crystalline forms of CDK inhibitors and pharmaceutically acceptable salts thereof, and uses thereof. [Background technology]

[0003] In recent years, tumors have surpassed cardiovascular disease as the leading cause of death worldwide, making research into antitumor drugs of great academic and practical significance. Because excessive activation and sustained cell proliferation are fundamental characteristics of tumors, inducing cell cycle arrest can effectively inhibit tumor growth. Cyclin-dependent kinases (CDKs) belong to the serine / threonine protein kinase family and are important kinases involved in cell cycle regulation. To date, 20 CDKs have been reported, which can be divided into two main categories based on their CDK function. One category of CDKs is involved in cell cycle regulation and primarily includes CDK1, CDK2, CDK4, and CDK6. The other category of CDKs is involved in transcription regulation and primarily includes CDK7, CDK8, CDK9, CDK10, and CDK11. In tumor cells, overexpression or overactivation of cyclins, inhibition of CDKi activity, and sustained activation of upstream mitogenic signals all lead to altered CDK activity. Dysregulation of CDK activity directly or indirectly leads to uncontrolled cell proliferation, genome instability (increased DNA mutations, chromosome deletions, etc.), and chromosome instability (changes in the number of chromosomes), and is involved in the development and progression of tumors. Because CDK activity is required for cell division and is often enhanced in tumor cells, CDKs have long been considered promising targets for drug research and development for tumors and other proliferative disorders.

[0004] Currently, the clinical application of many CDK4 / 6 inhibitors has fully verified their efficacy in targeting these targets. For example, Pfizer's Palbociclib, Eli Lilly's Abemaciclib, Novartis' Ribociclib, and Hengrui Pharmaceutical's Dalpiciclib are all used to treat hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2-) advanced or metastatic breast cancer.

[0005] Endocrine therapy is often the treatment of choice for patients with HR+ / HER2- advanced or metastatic breast cancer in the early stages. Common endocrine therapy drugs include anastrozole, letrozole, exemestane, tamoxifen, toremifene, fulvestrant, megestrol (commonly referred to as "megestrol acetate"), fluoxymesterone, and ethinyl estradiol. Patients with early-stage, intermediate- or high-risk disease can receive chemotherapy concomitantly. For patients with advanced or metastatic disease, the current first-line standard of care is a CDK4 / 6 inhibitor in combination with endocrine therapy. Many authoritative guidelines, both domestic and international, including the Breast Cancer Clinical Practice Guidelines (2022 edition) issued by the National Health Commission of China, the CSCO Breast Cancer Clinical Practice Guidelines (2022 edition), and the NCCN Breast Cancer Clinical Practice Guidelines (2022 edition), have all upgraded the evidence level for the combination of CDK4 / 6 inhibitors with endocrine therapy to Level I and recommended its clinical application in HR+ / HER2- advanced breast cancer.

[0006] International Publication WO2021139817A1 discloses a CDK inhibitor compound A, the structure of which is represented by the following formula (I): [ka]

[0007] In vitro studies have shown that the compound has excellent inhibitory effects against various CDK kinases and TRK kinases and excellent in vivo bioavailability, and is expected to be used in the treatment of various related diseases such as tumors. However, the crystalline forms of the compound and its pharmaceutically acceptable salts have not yet been systematically studied. Summary of the Invention

[0008] According to a first aspect, the present invention provides a crystalline form of Compound A or a pharmaceutically acceptable salt thereof, wherein the structure of Compound A is represented by Formula (I): [ka]

[0009] In some embodiments, the crystalline form of Compound A has an endothermic peak at 256±3°C in a DSC pattern.

[0010] In some embodiments, the present invention provides a crystalline form of Compound A. In some embodiments, the crystalline form of Compound A is anhydrous.

[0011] In some embodiments, the crystalline form of Compound A is crystalline form AI, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 5.2±0.2°, 11.4±0.2°, 15.7±0.2°, 20.2±0.2°, and 22.0±0.2°.

[0012] In some embodiments, the crystalline form of Compound A is crystalline form AI, and the X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 5.2±0.2°, 9.5±0.2°, 11.4±0.2°, 15.7±0.2°, 16.3±0.2°, 20.2±0.2°, and 22.0±0.2°.

[0013] In some embodiments, the crystalline form of Compound A is crystalline form AI, and the X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 5.2±0.2°, 9.5±0.2°, 10.4±0.2°, 11.4±0.2°, 12.8±0.2°, 14.1±0.2°, 15.7±0.2°, 16.3±0.2°, 18.6±0.2°, 20.2±0.2°, 22.0±0.2°, and 25.8±0.2°.

[0014] In some embodiments, the crystalline form of Compound A is crystalline form AI, and the peak positions and relative peak intensities of the diffraction peaks in the X-ray powder diffraction pattern, expressed as 2θ angles (°) using Cu-Kα radiation, are shown in Table 1 below.

[0015] [Table 1]

[0016] In some embodiments, the crystalline form of Compound A is crystalline form AI, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 1-1.

[0017] In some embodiments, the crystalline form of Compound A is crystalline form AI, and the DSC pattern of crystalline form AI has an endothermic peak at 256±3°C.

[0018] In some embodiments, the crystalline form of Compound A is crystalline form AI, and the DSC pattern of crystalline form AI is substantially as shown in Figures 1-3.

[0019] In some embodiments, the crystalline form of Compound A is crystalline form A-II, and its X-ray powder diffraction pattern, expressed in 2θ angles (°), using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 16.2±0.2°, 16.8±0.2°, 20.1±0.2°, and 20.7±0.2°.

[0020] In some embodiments, the crystalline form of Compound A is crystalline form A-II, and its X-ray powder diffraction pattern, expressed in degrees 2θ using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 16.2±0.2°, 16.8±0.2°, 17.7±0.2°, 20.1±0.2°, and 20.7±0.2°.

[0021] In some embodiments, the crystalline form of Compound A is crystalline form A-II, and the peak positions and relative peak intensities of the diffraction peaks in the X-ray powder diffraction pattern, expressed as 2θ angles (°) using Cu-Kα radiation, are shown in Table 2 below.

[0022] [Table 2]

[0023] In some embodiments, the crystalline form of Compound A is crystalline form A-II, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 2-1.

[0024] In some embodiments, the crystalline form of Compound A is crystalline form A-II, and crystalline form A-II is a solvate. In some embodiments, crystalline form A-II is a trifluoroethanol solvate of crystalline form of Compound A. In some embodiments, crystalline form A-II is a trifluoroethanol solvate of crystalline form of Compound A, and the molar ratio of Compound A to trifluoroethanol is 1:1.

[0025] In some embodiments, the crystalline form of Compound A is crystalline form A-II, and the DSC pattern of crystalline form A-II has endothermic peaks at 100 to 130°C and 256±3°C.

[0026] In some embodiments, the crystalline form of Compound A is crystalline form A-II, and the DSC pattern of crystalline form A-II is substantially as shown in Figure 2-3.

[0027] In some embodiments, the crystalline form of Compound A is crystalline form A-III, and its X-ray powder diffraction pattern, expressed in 2θ angles (°), using Cu-Kα radiation, has characteristic peaks at 6.6±0.2°, 13.2±0.2°, 16.5±0.2°, 20.1±0.2°, and 23.0±0.2°.

[0028] In some embodiments, the crystalline form of Compound A is crystalline form A-III, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 6.6±0.2°, 13.2±0.2°, 15.2±0.2°, 16.5±0.2°, 20.1±0.2°, 23.0±0.2°, and 28.2±0.2°.

[0029] In some embodiments, the crystalline form of Compound A is crystalline form A-III, and the peak positions and relative peak intensities of the diffraction peaks in an X-ray powder diffraction pattern expressed as 2θ angles (°) using Cu-Kα radiation are as shown in Table 3 below.

[0030] [Table 3]

[0031] In some embodiments, the crystalline form of Compound A is crystalline form A-III, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 3-1.

[0032] In some embodiments, the crystalline form of Compound A is crystalline form A-III, and crystalline form A-III is a solvate. In some embodiments, crystalline form A-III is a dimethyl sulfoxide solvate of crystalline form of Compound A. In some embodiments, crystalline form A-III is a dimethyl sulfoxide solvate of crystalline form of Compound A, and the molar ratio of Compound A to dimethyl sulfoxide is 1:1.

[0033] In some embodiments, the crystalline form of Compound A is crystalline form A-III, and the DSC pattern of crystalline form A-III has endothermic peaks at 100 to 150°C and 256±3°C.

[0034] In some embodiments, the crystalline form of Compound A is crystalline form A-III, and the DSC pattern of crystalline form A-III is substantially as shown in Figure 3-3.

[0035] In some embodiments, the crystalline form of Compound A is crystalline form A-IV, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 5.4±0.2°, 9.5±0.2°, 12.1±0.2°, 15.9±0.2°, and 20.7±0.2°.

[0036] In some embodiments, the crystalline form of Compound A is crystalline form A-IV, and its X-ray powder diffraction pattern, expressed in degrees 2θ using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 5.4±0.2°, 9.5±0.2°, 12.1±0.2°, 15.9±0.2°, 17.8±0.2°, 19.3±0.2°, and 20.7±0.2°.

[0037] In some embodiments, the crystalline form of Compound A is crystalline form A-IV, and the peak positions and relative peak intensities of the diffraction peaks in an X-ray powder diffraction pattern expressed as 2θ angles (°) using Cu-Kα radiation are as shown in Table 4 below.

[0038] [Table 4]

[0039] In some embodiments, the crystalline form of Compound A is crystalline form A-IV, and the X-ray powder diffraction pattern using Cu-Kα radiation is substantially as shown in Figure 4-1. In some embodiments, the crystalline form of Compound A is crystalline form A-IV, and the DSC pattern of crystalline form A-IV is substantially as shown in Figure 4-3.

[0040] In some embodiments, the pharmaceutically acceptable salt of Compound A in crystalline form is selected from sulfate, methanesulfonate, tartrate, benzenesulfonate, hydrochloride, p-toluenesulfonate, fumarate, citrate, and malate, preferably benzenesulfonate, hydrochloride, p-toluenesulfonate, fumarate, citrate, and malate, more preferably hydrochloride and benzenesulfonate.

[0041] In some embodiments, the crystalline form of Compound A is a pharmaceutically acceptable salt selected from the hydrochloride salt. In some embodiments, the crystalline form of Compound A is a hydrochloride hydrate. In some embodiments, the crystalline form of Compound A is a monohydrate of Compound A hydrochloride.

[0042] In some embodiments, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 8.9±0.2°, 14.3±0.2°, and 19.9±0.2°.

[0043] In some embodiments, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 8.9±0.2°, 12.3±0.2°, 14.3±0.2°, 15.4±0.2°, 16.8±0.2°, and 19.9±0.2°.

[0044] In some embodiments, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, and its X-ray powder diffraction pattern, expressed in 2θ angles (°) using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 8.3±0.2°, 8.9±0.2°, 9.5±0.2°, 12.3±0.2°, 14.3±0.2°, 15.0±0.2°, 15.4±0.2°, 16.8±0.2°, 19.1±0.2°, 19.9±0.2°, 20.6±0.2°, and 25.5±0.2°.

[0045] In some embodiments, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, and the X-ray powder diffraction pattern, expressed in 2θ angles (°) using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 8.3±0.2°, 8.9±0.2°, 9.5±0.2°, 11.1±0.2°, 12.3±0.2°, 14.3±0.2°, 15.0±0.2°, 15.4±0.2°, 16.8±0.2°, 17.4±0.2°, 19.1±0.2°, 19.9±0.2°, 20.6±0.2°, 21.6±0.2°, 25.2±0.2°, 25.5±0.2°, 26.6±0.2°, and 29.1±0.2°.

[0046] In some embodiments, the crystalline form of Compound A hydrochloride salt is crystalline form B, and the peak positions and relative peak intensities of the diffraction peaks in the X-ray powder diffraction pattern, expressed as 2θ angles (°) using Cu-Kα radiation, are as shown in Table 5 below.

[0047] [Table 5]

[0048] In some embodiments, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 5-1.

[0049] In some embodiments, the crystalline form of Compound A hydrochloride salt is crystalline form B, and the weight loss in the range of room temperature to 130° C. is about 4.4% in the TGA pattern.

[0050] In some embodiments, the crystalline form of Compound A hydrochloride salt is crystalline form B, and the DSC pattern has an endothermic peak at 253±3°C; in some embodiments, the crystalline form of Compound A hydrochloride salt is crystalline form B, and the DSC pattern has two endothermic peaks at 27±3°C and 253±3°C.

[0051] In some embodiments, the crystalline form B is a crystalline form of the hydrochloride salt monohydrate of Compound A, wherein the molar ratio of Compound A, hydrochloric acid, and water is 1:1:1.

[0052] In some embodiments, the crystalline form of Compound A is a pharmaceutically acceptable salt selected from benzenesulfonates. In some embodiments, the crystalline form of Compound A is an anhydrous benzenesulfonate. In some embodiments, the molar ratio of Compound A free base to benzenesulfonic acid in the crystalline form of Compound A is 1:1.

[0053] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and its X-ray powder diffraction pattern, expressed in 2θ angles (°), using Cu-Kα radiation, has characteristic peaks at 8.5±0.2°, 15.3±0.2°, 19.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°.

[0054] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 8.5±0.2°, 15.3±0.2°, 16.4±0.2°, 17.1±0.2°, 17.8±0.2°, 19.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°.

[0055] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and has an X-ray powder diffraction pattern expressed in 2θ angles (°) using Cu-Kα radiation, with characteristic peaks at 8.5±0.2°, 15.3±0.2°, 16.4±0.2°, 17.1±0.2°, 17.8±0.2°, 18.1±0.2°, 19.2±0.2°, 20.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°.

[0056] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and its X-ray powder diffraction pattern, expressed in 2θ angles (°), using Cu-Kα radiation, has characteristic peaks at 8.5±0.2°, 11.7±0.2°, 12.7±0.2°, 13.3±0.2°, 14.2±0.2°, 15.3±0.2°, 16.4±0.2°, 17.1±0.2°, 17.8±0.2°, 18.1±0.2°, 19.2±0.2°, 19.5±0.2°, 20.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°.

[0057] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and the peak positions and relative peak intensities of the diffraction peaks in an X-ray powder diffraction pattern expressed in 2θ angles (°) using Cu-Kα radiation are as shown in Table 6 below.

[0058] [Table 6]

[0059] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 6-1.

[0060] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and has an endothermic peak at 253±3° C. in a DSC pattern.

[0061] In some embodiments, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, having a DSC pattern substantially as shown in Figure 6-2.

[0062] In some embodiments, crystalline Form C is a crystalline form of the benzenesulfonic acid salt of Compound A, wherein the molar ratio of Compound A to benzenesulfonic acid is 1:1.

[0063] In some embodiments, the crystalline form of Compound A is a pharmaceutically acceptable salt selected from the p-toluenesulfonate salt. In some embodiments, the crystalline form of Compound A is a p-toluenesulfonate salt.

[0064] In some embodiments, the crystalline form of p-toluenesulfonate salt of Compound A is crystalline form DI, and its X-ray powder diffraction pattern, expressed in 2θ angles (°), using Cu-Kα radiation, has characteristic peaks at 9.1±0.2°, 13.3±0.2°, 15.8±0.2°, 19.6±0.2°, and 20.9±0.2°.

[0065] In some embodiments, the crystalline form of p-toluenesulfonate salt of Compound A is crystalline form DI, and has an X-ray powder diffraction pattern expressed in degrees 2θ (°) using Cu-Kα radiation, with characteristic peaks at 7.9±0.2°, 9.1±0.2°, 12.9±0.2°, 13.3±0.2°, 15.8±0.2°, 18.5±0.2°, 19.6±0.2°, and 20.9±0.2°.

[0066] In some embodiments, the crystalline form of p-toluenesulfonate salt of Compound A is crystalline form DI, and the peak positions and relative peak intensities of the diffraction peaks in an X-ray powder diffraction pattern expressed as 2θ angles (°) using Cu-Kα radiation are as shown in Table 7 below.

[0067] [Table 7]

[0068] In some embodiments, the crystalline form of p-toluenesulfonate salt of Compound A is crystalline form DI, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 7-1.

[0069] In some embodiments, the crystalline form of p-toluenesulfonate salt of Compound A is crystalline form DI, and exhibits a weight loss of about 4.2% in the range of room temperature to 150° C. in TGA.

[0070] In some embodiments, the crystalline form of p-toluenesulfonate salt of Compound A is crystalline form DI and the DSC pattern has a relatively broad endothermic peak at 167±3°C, and in some embodiments, the crystalline form of p-toluenesulfonate salt of Compound A is crystalline form DI and the DSC pattern has two relatively broad endothermic peaks at 49±3°C and 167±3°C.

[0071] In some embodiments, the crystalline form of p-toluenesulfonic acid salt of Compound A is crystalline form DI, having a DSC pattern substantially as shown in Figure 7-2.

[0072] In some embodiments, crystalline Form DI is a crystalline form of Compound A p-toluenesulfonic acid salt monohydrate, wherein the molar ratio of Compound A, p-toluenesulfonic acid, and water is 1:1:1.

[0073] In some embodiments, the crystalline form of Compound A's pharmaceutically acceptable salt is selected from a fumarate salt. In some embodiments, the molar ratio of fumaric acid to Compound A's free base in the crystalline form of Compound A's fumarate salt is 1:1. In some embodiments, the crystalline form of Compound A's fumarate salt is a hydrate. In some embodiments, the crystalline form of Compound A's fumarate salt is a monohydrate.

[0074] In some embodiments, the crystalline form of fumarate salt of Compound A is crystalline form E, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 13.2±0.2°, 16.6±0.2°, 19.9±0.2°, and 21.5±0.2°.

[0075] In some embodiments, the crystalline form of fumarate salt of Compound A is crystalline form E, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 4.1±0.2°, 8.4±0.2°, 12.5±0.2°, 13.2±0.2°, 13.9±0.2°, 16.6±0.2°, 19.9±0.2°, 21.5±0.2°, and 22.1±0.2°.

[0076] In some embodiments, the crystalline form of fumarate salt of Compound A is crystalline form E, and the peak positions and relative peak intensities of the diffraction peaks in an X-ray powder diffraction pattern expressed in 2θ angles (°) using Cu-Kα radiation are as shown in Table 8 below.

[0077] [Table 8]

[0078] In some embodiments, the crystalline form of fumarate salt of Compound A is crystalline form E, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 9-1.

[0079] In some embodiments, the crystalline form of fumarate salt of Compound A is crystalline form E, and the DSC pattern has endothermic peaks at 222±3°C, 230±3°C, and 240±3°C. In some embodiments, the crystalline form of fumarate salt of Compound A is crystalline form E, and the DSC pattern has endothermic peaks at 159±3°C, and endothermic peaks at 32±3°C, 222±3°C, 230±3°C, and 240±3°C.

[0080] In some embodiments, the crystalline form of fumarate salt of Compound A is crystalline form E, having a DSC pattern substantially as shown in Figure 9-2.

[0081] In some embodiments, the crystalline form E is a crystalline form of Compound A fumarate monohydrate, wherein the molar ratio of Compound A, fumaric acid, and water is 1:1:1.

[0082] In some embodiments, the crystalline form of Compound A is a pharmaceutically acceptable salt selected from the group consisting of a citrate salt, a crystalline form of Compound A, ...

[0083] In some embodiments, the crystalline form of citrate salt of Compound A is crystalline form F, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 10.1±0.2°, 15.5±0.2°, 17.1±0.2°, 19.4±0.2°, 20.0±0.2°, and 21.5±0.2°.

[0084] In some embodiments, the crystalline form of citrate salt of Compound A is crystalline form F, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 10.1±0.2°, 11.1±0.2°, 12.5±0.2°, 15.5±0.2°, 17.1±0.2°, 18.4±0.2°, 19.4±0.2°, 20.0±0.2°, 21.5±0.2°, 23.0±0.2°, and 24.2±0.2°.

[0085] In some embodiments, the crystalline form of citrate salt of Compound A is crystalline form F, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 10.1±0.2°, 11.1±0.2°, 12.5±0.2°, 13.0±0.2°, 14.1±0.2°, 15.5±0.2°, 16.6±0.2°, 17.1±0.2°, 17.5±0.2°, 18.4±0.2°, 19.4±0.2°, 20.0±0.2°, 20.5±0.2°, 21.5±0.2°, 23.0±0.2°, 24.2±0.2°, and 24.7±0.2°.

[0086] In some embodiments, the crystalline form of citrate salt of Compound A is crystalline form F, and the peak positions and relative peak intensities of the diffraction peaks in an X-ray powder diffraction pattern expressed in 2θ angles (°) using Cu-Kα radiation are as shown in Table 9 below.

[0087] [Table 9]

[0088] In some embodiments, the crystalline form of citrate salt of Compound A is crystalline form F, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 10-1.

[0089] In some embodiments, the crystalline form of citrate salt of Compound A is crystalline form F, having a DSC pattern with three endothermic peaks at 30±3°C, 198±3°C, and 248±3°C.

[0090] In some embodiments, the crystalline form of citrate salt of Compound A is crystalline form F, having a DSC pattern substantially as shown in Figure 10-2.

[0091] In some embodiments, crystalline form F is a crystalline form of Compound A citrate hydrate, wherein the molar ratio of Compound A to citric acid is 1:1.

[0092] In some embodiments, the crystalline form of Compound A is a pharmaceutically acceptable salt selected from the malate salt. In some embodiments, the molar ratio of malic acid to Compound A free base in the crystalline form of Compound A malate is 1:1. In some embodiments, the crystalline form of Compound A malate is a hydrate. In some embodiments, the crystalline form of Compound A malate is a monohydrate.

[0093] In some embodiments, the crystalline form of Compound A malate is crystalline form G, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 4.0±0.2°, 11.4±0.2°, 12.4±0.2°, 19.6±0.2°, 20.4±0.2°, 21.1±0.2°, and 22.5±0.2°.

[0094] In some embodiments, the crystalline form of Compound A malate salt is crystalline form G, and its X-ray powder diffraction pattern, expressed in degrees 2θ (°), using Cu-Kα radiation, has characteristic peaks at 4.0±0.2°, 9.8±0.2°, 10.5±0.2°, 11.4±0.2°, 12.4±0.2°, 13.2±0.2°, 14.3±0.2°, 15.9±0.2°, 17.0±0.2°, 19.0±0.2°, 19.6±0.2°, 20.4±0.2°, 21.1±0.2°, 22.5±0.2°, 24.1±0.2°, and 25.4±0.2°.

[0095] In some embodiments, the crystalline form of Compound A malate is crystalline form G, and the peak positions and relative peak intensities of the diffraction peaks in an X-ray powder diffraction pattern expressed in 2θ angles (°) using Cu-Kα radiation are as shown in Table 10 below.

[0096] [Table 10]

[0097] In some embodiments, the crystalline form of Compound A malate is crystalline form G, having an X-ray powder diffraction pattern using Cu-Kα radiation substantially as shown in Figure 11-1.

[0098] In some embodiments, the crystalline form of Compound A malate salt is crystalline form G, having a DSC pattern with endothermic peaks at 57±3°C, 175±3°C, 191±3°C, and 238±3°C.

[0099] In some embodiments, the crystalline form of Compound A malate salt is crystalline form G, having a DSC pattern substantially as shown in Figure 11-2.

[0100] In some embodiments, the crystalline form G is a monohydrate of the malic acid salt of Compound A, wherein the molar ratio of Compound A, malic acid, and water is 1:1:1.

[0101] According to a second aspect, the present invention further provides a pharmaceutical composition comprising a crystalline form of Compound A, or a pharmaceutically acceptable salt thereof, as described above. In some embodiments, the present invention provides a pharmaceutical composition comprising one or more of crystalline Form AI, Form A-II, Form A-III, Form A-IV, Form B, Form C, Form DI, Form E, Form F, and Form G, as described above, and optionally one or more pharmaceutically acceptable excipients. Preferably, the pharmaceutical composition comprises one or more of crystalline Form AI, Form B, and Form C, as described above, and optionally one or more pharmaceutically acceptable excipients.

[0102] According to a third aspect, the present invention further provides the use of the crystalline form of Compound A or a pharmaceutically acceptable salt thereof as described above in the manufacture of an antitumor drug. In some embodiments, the present invention provides the use of crystalline form AI, crystalline form A-II, crystalline form A-III, crystalline form A-IV, crystalline form B, crystalline form C, crystalline form DI, crystalline form E, crystalline form F, and crystalline form G, or a pharmaceutical composition as described above, in the manufacture of an antitumor drug.

[0103] According to a fourth aspect, the present invention further provides a method for treating a tumor with a crystalline form of Compound A or a pharmaceutically acceptable salt thereof, as described above, comprising administering a therapeutically effective amount of the crystalline form of Compound A or a pharmaceutically acceptable salt thereof to a subject or patient in need of such treatment. In some embodiments, the present invention provides a method for treating a tumor, comprising administering a therapeutically effective amount of crystalline Form AI, Form A-II, Form A-III, Form A-IV, Form B, Form C, Form DI, Form E, Form F, or Form G, or a pharmaceutical composition, as described above, to a subject or patient in need of such treatment.

[0104] According to a fifth aspect, the present invention further provides a crystalline form of Compound A or a pharmaceutically acceptable salt thereof as described above for treating tumors. In some embodiments, the present invention further provides crystalline form AI, crystalline form A-II, crystalline form A-III, crystalline form A-IV, crystalline form B, crystalline form C, crystalline form DI, crystalline form E, crystalline form F, and crystalline form G, or a pharmaceutical composition thereof, as described above for treating tumors.

[0105] According to a sixth aspect, the present invention further provides the use of the crystalline form of Compound A, or a pharmaceutically acceptable salt thereof, as described above, in tumor therapy. In some embodiments, the present invention further provides the use of crystalline form AI, crystalline form A-II, crystalline form A-III, crystalline form A-IV, crystalline form B, crystalline form C, crystalline form DI, crystalline form E, crystalline form F, and crystalline form G, or a pharmaceutical composition thereof, as described above, in tumor therapy.

[0106] According to each of the above aspects, In some embodiments, the tumor is selected from a sensitive tumor or a drug-resistant tumor. Preferably, the sensitive tumor or drug-resistant tumor is selected from a solid tumor or a hematological tumor. Preferably, the solid tumor is selected from fibrosarcoma, salivary gland cancer, liver cancer, colon cancer, bladder cancer, pharyngeal cancer, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, renal cancer, pancreatic cancer, skin cancer, lymphoma, gastric cancer, multiple myeloma, brain tumor, lung cancer, and malignant melanoma, and the hematological tumor is selected from acute myeloid leukemia. More preferably, the sensitive tumor or drug-resistant tumor is selected from breast cancer, lung cancer, colon cancer, liver cancer, ovarian cancer, malignant melanoma, and acute myeloid leukemia. In some embodiments, the breast cancer is selected from HR+ / HER2- breast cancer, triple-negative breast cancer, and HR- / HER2+ breast cancer, the acute myeloid leukemia is selected from myelomonocytic leukemia, and the lung cancer is selected from non-small cell lung cancer.

[0107] In some embodiments, the tumor is selected from a drug-resistant tumor, the drug-resistant tumor is selected from a primary drug-resistant tumor or an acquired drug-resistant tumor.

[0108] In some embodiments, the drug-resistant tumor is selected from a CDK4 / 6 inhibitor-resistant tumor, and further selected from a CDK4 / 6 inhibitor-primary resistant tumor or a CDK4 / 6 inhibitor-acquired resistant tumor, and the CDK4 / 6 inhibitor is selected from palbociclib, abemaciclib, ribociclib, and dalpiciclib, preferably palbociclib.

[0109] In some embodiments, the CDK4 / 6 inhibitor-resistant tumor is selected from breast cancer and liver cancer, more preferably from palbociclib-resistant breast cancer and liver cancer, and even more preferably from palbociclib-resistant breast cancer or liver cancer, or palbociclib-acquired-resistant breast cancer or liver cancer. The palbociclib-primary-resistant breast cancer and palbociclib-acquired-resistant breast cancer are selected from palbociclib-resistant or acquired-resistant HR+ / HER2- breast cancer, triple-negative breast cancer, and HR- / HER2+ breast cancer.

[0110] The crystalline form of Compound A or its pharmaceutically acceptable salt according to the present invention may be administered as a monotherapy or in combination with other clinically / pharmaceutically acceptable antitumor drugs, biological therapies, radiotherapy, and / or herbal therapies. In some embodiments, the other antitumor drugs are selected from endocrine therapy drugs such as anastrozole, letrozole, exemestane, tamoxifen, toremifene, fulvestrant, megestrol, fluoxymesterone, and ethinylestradiol, and preferably selected from fulvestrant.

[0111] Accordingly, the present invention provides the use of a crystalline form of Compound A or a pharmaceutically acceptable salt thereof in combination with another clinically / pharmaceutically acceptable drug in the manufacture of an antitumor drug. The present invention also provides an antitumor drug comprising a crystalline form of Compound A or a pharmaceutically acceptable salt thereof and another clinically / pharmaceutically acceptable drug. The other drug and the crystalline form of Compound A or a pharmaceutically acceptable salt thereof may be contained in the same dosage unit or in different dosage units to be packaged together, e.g., in a kit product. In some embodiments, the other drug is selected from endocrine therapy drugs, such as anastrozole, letrozole, exemestane, tamoxifen, toremifene, fulvestrant, megestrol, fluoxymesterone, and ethinyl estradiol, preferably fulvestrant.

[0112] The inventors have studied the crystalline state of Compound A and its pharmaceutically acceptable salts. They found that Compound A could not be reacted with maleic acid or phosphoric acid to form crystalline salts, while the salt samples obtained with sulfuric acid, methanesulfonic acid, and tartaric acid had low crystallinity and could not completely react with succinic acid. The hydrochloride, p-toluenesulfonate, citrate, fumaric acid, and malate salts were solvates / hydrates, while only the benzenesulfonate salt was anhydrous. Based on the results of solid-state characterization, the hydrochloride crystalline form B (hydrate) and the benzenesulfonate crystalline form C (anhydrous) were the predominant salt forms.

[0113] Further comparison of the solubility and stability of the hydrochloride salt form B (hydrate), the benzenesulfonate salt form C (anhydrous), and the free base form AI revealed that the free base form AI had low aqueous solubility, while the two salt forms had significantly higher aqueous solubility. However, in the other three biologically relevant media, the solubility of the free base form AI was not significantly different from that of the other two salt forms, and all had solubilities greater than 5 mg / mL in SGF. Solid-state stability results indicated that the benzenesulfonate salt form C could be converted to a hydrate under 92.5% RH conditions. The free base form AI, the hydrochloride salt form B, and the benzenesulfonate salt form C all maintained consistent physical and chemical stability after 7 days of storage under three conditions: 60°C, 92.5% RH, and 40°C / 75% RH.

[0114] Preclinical studies have shown that crystalline Compound A or its pharmaceutically acceptable salts can significantly inhibit the growth of various tumor cells, including those from breast cancer, colon cancer, ovarian cancer, malignant melanoma, liver cancer, lung cancer, and acute myeloid leukemia, and have excellent inhibitory effects against palbociclib-resistant (primary resistance and acquired resistance) breast cancer and liver cancer.

[0115] Therefore, the crystalline form of Compound A or a pharmaceutically acceptable salt thereof has excellent clinical applicability in the treatment of progressive malignant tumors (advanced malignant tumors), and provides a new drug option for the treatment of CDK4 / 6 inhibitor-resistant tumors.

[0116] [Definition and Explanation] Unless otherwise specified, the following terms and expressions used herein have the following meanings: A particular expression or term, unless specifically defined, is not to be deemed indefinite or ambiguous, but is to be understood as having its general meaning.

[0117] The crystalline form of Compound A or a pharmaceutically acceptable salt thereof referred to in the present invention includes anhydrous, solvent-free, hydrated, solvated and co-crystalline forms of Compound A or a pharmaceutically acceptable salt thereof.

[0118] The "room temperature" mentioned above is the temperature of room temperature in the general sense in this field, which is generally 10 to 30°C, and preferably 25°C±5°C.

[0119] In the context of X-ray powder diffraction patterns, the terms "substantially" or "substantially as shown in the figure" refer to a substantially pure specific 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 X-ray powder diffraction pattern appear in the pattern. Furthermore, as the content of a specific crystalline form in a product gradually decreases, some of the diffraction peaks belonging to that crystalline form in the X-ray powder diffraction pattern may decrease due to factors such as instrument sensitivity. It is also well known in the field of crystallography that for any given crystalline form, there may be slight errors in peak positions. For example, peak positions may fluctuate due to temperature changes during sample analysis, sample movement, or instrument calibration, resulting in a measurement error of approximately ±0.3°, typically about ±0.2°, of 2θ values. Therefore, when identifying various crystalline structures, such errors should be taken into consideration, and the terms "substantially" or "substantially as shown in the figure" are intended to encompass such differences in diffraction peak positions, and such differences are within ±0.3°, preferably ±0.2°.

[0120] In a DSC pattern or a TGA pattern, the term "substantially" or "substantially as shown in the figure" means that, in the case of the same crystalline form of the same compound, the error in the thermal transition onset temperature, the peak temperature of the endothermic peak, the peak temperature of the exothermic peak, the melting point, the onset temperature of weight loss, or the end temperature of weight loss, etc., may be typically ±5°C, and usually ±3°C, in successive analyses.

[0121] The term "tumor" includes benign tumors, malignant tumors, and borderline tumors, of which malignant tumors are collectively referred to as cancer.

[0122] As used herein, the term "prevention," when applied to a disease or disorder (also called a "symptom") (e.g., cancer), refers to the ability of a compound or drug (e.g., a combination product claimed for protection in the present invention) to reduce the frequency of or delay the onset of symptoms of a medical disorder in a subject compared to a subject not administered the compound or drug.

[0123] As used herein, the term "treatment" refers to alleviating, alleviating, or ameliorating the symptoms of a disease or disorder, ameliorating underlying metabolic symptoms, inhibiting a disease or condition, e.g., arresting the progression of a disease or disorder, relieving a disease or disorder, attenuating a disease or disorder, alleviating the pathology of a disease or disorder, or preventing the symptoms of a disease or disorder.

[0124] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refers to a carrier or excipient that has no appreciable irritating effect on an organism and does not impair the biological activity and performance of the active compound.

[0125] The terms "subject" and "patient" include, but are not limited to, mammals (e.g., mice, rats, cats, monkeys, dogs, horses, pigs) and humans.

[0126] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present invention or salts thereof with a pharmaceutically acceptable excipient. The pharmaceutical composition of the present invention can be prepared by combining the compound of the present invention with a suitable pharmaceutically acceptable excipient. The pharmaceutical composition of the present invention can be prepared by a method well known in the art, such as a general mixing method, dissolving method, granulating method, dragee-making method, pulverizing method, emulsifying method, or lyophilizing method.

[0127] The term "therapeutically effective amount" refers to a non-toxic but sufficient amount of a drug or pharmacologically active agent to achieve the desired effect. The determination of an effective amount varies from person to person, depending on the age and general condition of the individual (subject), and also on the specific active substance. The appropriate effective amount for each regimen can be determined by one skilled in the art based on routine testing.

[0128] All solvents used in this invention are commercially available and can be used without further purification.

[0129] In order to provide a more concise explanation, the term "about" is not used for some quantitative data in this specification. Regardless of whether the term "about" is explicitly used, any numerical value given in this specification refers not only to the actual given value (predetermined value), but also to an approximation of the given value that can be reasonably estimated by a person skilled in the art, including equivalent values ​​and approximate values ​​resulting from experimental conditions and / or measurement conditions for such a given value. The above-mentioned approximation is preferably ±20%, ±15%, ±10%, ±8%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% based on the given value. [Brief explanation of the drawings]

[0130] [Figure 1-1] FIG. 1 shows the XRPD pattern of the free base crystalline form AI of Compound A. [Figure 1-2] FIG. 1 shows the TGA pattern of the free base crystalline form AI of Compound A. [Figure 1-3]FIG. 1 shows the DSC pattern of the free base crystalline form AI of Compound A. [Figure 2-1] FIG. 1 shows the XRPD pattern of free base crystalline form A-II of Compound A. [Figure 2-2] FIG. 1 shows the TGA pattern of the free base crystalline form A-II of Compound A. [Figure 2-3] FIG. 1 shows the DSC pattern of the free base crystalline form A-II of Compound A. [Figure 3-1] FIG. 1 shows the XRPD pattern of the free base crystalline form A-III of Compound A. [Figure 3-2] FIG. 1 shows the TGA pattern of the free base crystalline form A-III of Compound A. [Figure 3-3] FIG. 1 shows the DSC pattern of the free base crystalline form A-III of Compound A. [Figure 4-1] FIG. 1 shows the XRPD pattern of free base crystalline form A-IV of Compound A. [Figure 4-2] FIG. 1 shows the TGA pattern of the free base crystalline form A-IV of Compound A. [Figure 4-3] FIG. 1 shows the DSC pattern of the free base crystalline form A-IV of Compound A. [Figure 5-1] FIG. 1 shows the XRPD pattern of crystalline form B of the hydrochloride salt of Compound A. [Figure 5-2] FIG. 2 shows the TGA and DSC patterns of crystalline form B of the hydrochloride salt of Compound A. [Figure 6-1] FIG. 1 shows an XRPD pattern of crystalline Form C of the benzenesulfonate salt of Compound A. [Figure 6-2] FIG. 1 shows the TGA and DSC patterns of crystalline Form C of the benzenesulfonate salt of Compound A. [Figure 7-1] FIG. 1 shows an XRPD pattern of p-toluenesulfonate salt crystalline form DI of Compound A. [Figure 7-2] FIG. 2 shows the TGA and DSC patterns of p-toluenesulfonate crystalline form DI of Compound A. [Figure 8-1]FIG. 1 shows an XRPD pattern of crystalline form D-II of the p-toluenesulfonate salt of Compound A. [Figure 8-2] FIG. 2 shows the TGA and DSC patterns of crystalline form D-II of the p-toluenesulfonate salt of Compound A. [Figure 9-1] FIG. 1 shows the XRPD pattern of crystalline Form E of the fumarate salt of Compound A. [Figure 9-2] FIG. 2 shows the TGA and DSC patterns of crystalline Form E of the fumarate salt of Compound A. [Figure 10-1] FIG. 1 shows the XRPD pattern of crystalline Form F of the citrate salt of Compound A. [Figure 10-2] FIG. 2 shows the TGA and DSC patterns of crystalline Form F of the citrate salt of Compound A. [Figure 11-1] FIG. 1 shows the XRPD pattern of crystalline Form G of the malate salt of Compound A. [Figure 11-2] FIG. 2 shows the TGA and DSC patterns of crystalline Form G of the malate salt of Compound A. DETAILED DESCRIPTION OF THE INVENTION

[0131] The drugs, reagents, etc. in the following specific embodiments can all be purchased commercially or prepared by conventional methods.

[0132] The present invention will be further described below with reference to specific examples. It should be understood that these examples are merely illustrative of the present invention and do not limit the scope of the present invention. In the following examples, experimental methods for which specific conditions are not specified are generally carried out according to general conditions or conditions suggested by the manufacturer. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the method of the present invention. The preferred methods and materials described herein are merely illustrative.

[0133] Detection Method 1. X-ray powder diffraction (XRPD) Equipment:Bruker D8 Advance Diffractometer Detection method: Place the sample flat on a non-reflective sample plate and gently press the sample flat to detect. Specific conditions: Cu-Kα radiation, tube voltage 40 kV, tube current 40 mA, 2θ scanning range 3-40° or 3-30°, scanning step 0.02°, exposure time 0.2 seconds.

[0134] 2. Differential thermal analysis scanning (DSC) Equipment: TA Instruments Q200 DSC Detection method: Place 0.5 to 5 mg of sample into a perforated aluminum plate, heat it at a rate of 10 °C / min up to the final temperature, and purge the furnace with nitrogen at a rate of 50 mL / min.

[0135] 3.Thermogravimetric analysis (TGA) Equipment: TA Instruments Q500 TGA Detection method: Place 1-10 mg of sample on a balanced aluminum sample plate with an opening and automatically weigh it in the TGA furnace. The sample is heated to the final temperature at a rate of 10 °C / min, and the nitrogen purge rate in the sample chamber is 25 mL / min, and the nitrogen purge rate in the balance chamber is 40 mL / min.

[0136] 4. Dynamic Water Sorption (DVS) Equipment: TA Instruments Q5000 SA Detection method: Place 20-50 mg of sample into a balanced (deducting its own weight) sample basket, automatically weigh the sample, and perform DVS measurement on the sample according to the parameters in Table 7. The step time is 60 min, the equilibration time for each RH% is 1 hour, the total air flow rate is 200 sccm, and the test is performed in gradient mode. The humidity change is from 0% to 90% to 0%, with a humidity change of 10% for each gradient in the 0% to 90% range. The gradient endpoint is determined by the dm / dt method, and is reached when dm / dt is maintained at less than 0.01% for 10 minutes.

[0137] 5.Nuclear magnetic resonance apparatus (1H-NMR) 5.1 Study of free base crystal forms Detection method: 1H-NMR patterns are obtained using a Bruker Ascend 500 or WNMR-I 400 nuclear magnetic resonance spectrometer, with broadband excitation, a single pulse with a spectral width of 30 ppm, 16 scans at a 30° excitation angle, digital orthogonal detection, temperature control at 298 K, and deuteration reagent chloroform.

[0138] 5.2 Salt formation test Detection method: 1H-NMR patterns were obtained using a Bruker 400MHz nuclear magnetic resonance spectrometer. DMSO-d6 was used as the solvent, and broadband excitation was performed with a single pulse of 30 ppm spectral width, four scans at a 30° excitation angle, digital quadrature detection, and the temperature was controlled at 295K.

[0139] 6. Solubility Measurement At room temperature, a known amount of sample is weighed out, and the solvent is added to the sample in portions, stirred or sonicated to aid dissolution until the sample is visually clear, and the amount of solvent consumed is recorded. If the sample still does not clear at a particular concentration, its solubility is expressed as "<" the particular concentration.

[0140] JPEG2026502614000014.jpg55154

[0141] Production Example 1: Production of Compound A It is produced with reference to the method described in Example 2 of WO2021139817A1. The specific method is as follows.

[0142] [ka]

[0143] Compound 1S-a (500 mg, 1.84 mmol), N-methylpiperidone (2S-a, 416 mg, 3.68 mmol), and two drops of acetic acid were added to methanol (15 mL) and stirred at room temperature for 1 hour. Sodium cyanoborohydride (231 mg, 3.68 mmol) was then added, and the reaction mixture was stirred at room temperature overnight. The completion of the reaction was monitored by TLC. Methanol was removed from the reaction mixture under reduced pressure, followed by the addition of water (10 mL) and extraction with dichloromethane (3 × 20 mL). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography (dichloromethane:methanol = 30:1) to obtain compound 2S-b (500 mg) as a yellow solid, which was used directly in the next step.

[0144] Compound 2S-b (500 mg, 1.50 mmol) and palladium-carbon catalyst (10%, 70 mg) were added to methanol (15 mL) at room temperature. The reaction mixture was stirred at room temperature under a hydrogen gas atmosphere at 1 atm for 3 hours. Completion of the reaction was monitored by TLC. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane:methanol=20:1) to obtain compound 2S-c (280 mg, yield 62%) as a brown solid.

[0145] Compound 2S-c (280 mg, 0.93 mmol) and Ib (310 mg, 0.93 mmol) were dissolved in toluene (8 mL). The reaction mixture was stirred at 90 °C for 3 hours. Completion of the reaction was monitored by TLC. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to give compound 2S (180 mg, 34% yield) as a yellow solid.

[0146] 1 H NMR(500MHz,DMSO-d6)δ9.86(bs,1H), 8.90(s,1H), 7.16(d,J=2.1Hz,1H), 7.06(d,J=8.9Hz,1H), 6.81(d,J=8.9Hz,1H), 5.91-5.70(m,1H), 4.23(dd,J=10.5,2.4Hz,1H), 3.89(dd,J=10.5,9.0Hz,1 H), 3.69(d,J=11.3Hz,1H), 3.00-2.88(m,3H), 2.79(d,J=11.2Hz,2H), 2.59-2.53(m,1H), 2.41(s, 3H), 2.34-2.16(m,7H), 2.14(s,3H), 1.94-1.73(m,9H), 1.62-1.51(m,2H), 1.48-1.36(m,2H)ppm. MS m / z 572.8[M+H] + .

[0147] The compound 2S is the compound A described in the present invention. Upon detection, the yellow solid compound is amorphous.

[0148] Example 1: Preparation of free base crystalline form AI of Compound A Approximately 5 g of the sample obtained in Preparation Example 1 was taken, and 100 mL of n-butanol and 10 mL of anisole were added. The mixture was heated to 105±5°C and stirred to dissolve. The mixture was cooled to 85±5°C and crystallized for 1.5±0.5 hours, and continued to cool to 15±5°C and crystallized for 2.0±0.5 hours. The mixture was suction filtered, washed with 10 mL of n-butanol and 10 mL of n-heptane, and the filter cake was vacuum dried at 60±5°C for 4±2 hours to obtain Compound A free base crystalline form AI.

[0149] The solid was taken and subjected to XRPD characterization. The crystalline solid was designated as crystalline form AI. The XRPD characterization is shown in Figure 1-1, and the characterization data is shown in Table 11. The TGA results (see Figure 1-2) showed a weight loss of 0.30% from room temperature to 150°C, suggesting that the crystalline form is anhydrous, with a decomposition temperature of approximately 342°C. The DSC pattern (see Figure 1-3) showed a melting point of approximately 256°C. The DVS test results showed a weight change of approximately 0.17% from 0% RH to 90% RH, indicating that the crystalline form is not hygroscopic.

[0150] [Table 11]

[0151] As a result of the inventors' systematic research into the preparation of the free base crystalline form AI, other exemplary preparation methods include the following. JPEG2026502614000017.jpg116159

[0152] Example 2: Preparation of free base crystalline form A-II of Compound A (trifluoroethanol solvate of Compound A) Approximately 20 mg of the sample obtained in Preparation 1 was taken, clarified by adding 1.0 mL of trifluoroethanol and 0.5 mL of tetrahydrofuran, and rapidly spun to dryness at room temperature to obtain Compound A free base crystalline Form A-II.

[0153] The resulting crystalline solid was sampled and subjected to XRPD characterization (see Figure 2-1). See Table 12 for characterization data. The resulting crystalline solid is designated free base Form A-II. TGA (see Figure 2-2) indicates that the form has a weight loss of 16.9% before 150°C and a decomposition temperature of 343°C, suggesting that the form is a trifluoroethanol solvate of Compound A (containing 1 trifluoroethanol, theoretically 14.9%). DSC (see Figure 2-3) indicates that the form has a desolvation peak between 100°C and 130°C and a melting point of approximately 256°C (the melting point of Form AI).

[0154] [Table 12]

[0155] Example 3: Preparation of free base crystalline form A-III of Compound A (dimethyl sulfoxide solvate of Compound A) Approximately 30 mg of the sample obtained in Preparation Example 1 was taken, and 5.0 mL of dimethyl sulfoxide was added thereto. The mixture was clarified at 70°C, transferred to room temperature, and stirred for 2 hours to precipitate a solid. The solid was then centrifuged and vacuum dried overnight at 40°C to obtain crystalline Form A-III of the free base of Compound A.

[0156] A sample was taken for XRPD characterization (see Figure 3-1). See Table 13 for characterization data. The resulting crystalline solid is designated free base Form A-III. TGA testing (see Figure 3-2) indicates that the form has a weight loss of 12.4% before 150°C and a decomposition temperature of 342°C, suggesting that Form A-III is a dimethyl sulfoxide solvate of Compound A (containing 12.0% dimethyl sulfoxide, theoretically). DSC studies (see Figure 3-3) indicate that the form has a desolvation peak between 100°C and 150°C and a melting point of approximately 256°C (the melting point of free base Form AI).

[0157] [Table 13]

[0158] Example 4: Free base crystalline form A-IV of Compound A An appropriate amount of the amorphous sample was heated to 160° C. on a hot stage, kept at this temperature for 5 minutes, and then cooled to room temperature to obtain Compound A free base crystalline Form A-IV.

[0159] Sampling was performed for XRPD characterization (see Figure 4-1) and the resulting crystalline solid was designated Form A-IV, based on the characterization data in Table 14. The TGA pattern (see Figure 4-2) indicates that the form has a weight loss of 0.8% before 150°C and a decomposition temperature of 348°C. The DSC pattern (see Figure 4-3) indicates a melting point of approximately 255°C.

[0160] [Table 14]

[0161] Example 5: Compound A Hydrochloride Monohydrate Approximately 350 mg of compound A was weighed into a reaction flask, 7 mL of methanol was added, and the mixture was stirred at room temperature to obtain a suspension. After adding 1.05 equivalents of 1 M aqueous hydrochloric acid, the reaction mixture remained a suspension and was stirred overnight. The resulting solid was filtered and then dried under vacuum at 50 °C for 6 hours to obtain the above product (yield 75%, product purity 99.31%).

[0162] The IC analysis of the product (Table 15) shows that the molar ratio of the free base of Compound A to hydrochloric acid is 1:1 (theoretical chloride ion content in the hydrochloride salt: 5.8%). Therefore, in this experiment, a crystalline form of the hydrochloride salt of Compound A (designated as crystalline form B) was successfully prepared.

[0163] [Table 15]

[0164] The PLM and XRPD results (Figure 5-1) indicate that the sample is a highly crystalline, irregular crystal with small, aggregated particles. See Table 16 for characterization data. TGA revealed a weight loss of approximately 4.4% between room temperature and 130°C. DSC revealed two endothermic peaks at 27°C and 253°C, corresponding to the solvent loss and melting / decomposition peaks, respectively (see Figure 5-2). NMR revealed no significant residual organic solvent. Therefore, the weight loss was due to water loss, and the hydrochloride crystals are a monohydrate (theoretical water content of a monohydrate: 2.9%), defining them as hydrochloride crystalline Form B of Compound A.

[0165] [Table 16]

[0166] DVS measurement of the hydrochloride crystalline form B showed that the DVS curve of the sample was reversible, and the water content changed with changes in humidity. The moisture absorption of the sample ranged from 2.4% to 5.0% in the range of 10% to 90% RH, and the XRPD pattern of the sample did not change after DVS measurement.

[0167] The hydrochloride salt was prepared in different solvent systems with different acid equivalent dosages, as shown in Table 17. In addition to the presence of some free base crystalline form AI in the sample obtained in the acetone / water (4 / 1) system, the same XRPD pattern (shown in Figure 5-1) was obtained in all other experiments, and the patterns of the samples obtained with 2.1 and 5 equivalents of hydrochloric acid dosages were consistent with the XRPD pattern of the sample obtained with 1 equivalent of acid dosage, but the sample obtained with 5 equivalents dosage had slightly lower crystallinity and some impurity peaks were detected in the NMR pattern.

[0168] Therefore, the free base of Compound A was reacted with hydrochloric acid in a 1:1 ratio in methanol, acetone, and acetone / water systems, and 5 equivalents of acid were administered. As a result, even if the acid ratio was increased, it was not possible to obtain a hydrochloride salt with a different ratio, and there was a risk of decomposition.

[0169] [Table 17]

[0170] Example 6: Benzenesulfonate salt of Compound A The benzenesulfonate salt crystalline form C was prepared using methanol as a solvent. The specific experimental process and characterization results are as follows:

[0171] Approximately 300 mg of compound A was weighed into a reaction flask, and 6 mL (20 V) of methanol was added. The mixture was stirred at room temperature to obtain a suspension. 1.05 equivalents of solid benzenesulfonic acid was added, and the reaction mixture immediately became clear. After stirring for 1 hour, no solid precipitated. 30 mL of ethyl acetate was added, and the mixture was stirred at room temperature overnight. A solid precipitated. The resulting solid was filtered and dried in vacuo at 50°C for 6 hours to obtain the benzenesulfonate salt of compound A (yield: approximately 56%, product purity: 98.79%).

[0172] PLM and XRPD (Figure 6-1) indicate that the sample is irregularly crystalline, with small, aggregated particles. See Table 18 below for XRPD characterization data. As shown in Figure 6-2, TGA measurements show a weight loss of approximately 0.4% between room temperature and 100°C, which is attributed to solvent loss. DSC shows an endothermic melting peak at 235°C. NMR indicates that the molar ratio of free base to benzenesulfonic acid in the sample is 1:1. Therefore, benzenesulfonate salt crystalline form C of Compound A is an anhydrous crystalline form.

[0173] [Table 18]

[0174] DVS measurements showed that the sample absorbed only 1.2% moisture at 80% RH, but the moisture absorption rate increased sharply from 1.2% to 7% between 80% and 90% RH. During desorption, moisture was removed again at 60% RH, and at 40% RH, moisture was substantially removed. Therefore, under high humidity conditions, benzenesulfonate salt exists as a hydrate crystalline form, and Form C may convert to a hydrate, which may then convert back to the anhydrous crystalline form during desorption. Furthermore, DVS data indicates that the hydrate converted from Form C of benzenesulfonate salt dehydrates below 60% RH, and completely dehydrates at 40% RH, converting to the anhydrous crystalline form of benzenesulfonate salt. Therefore, the hydrate is stable within a humidity range of 60-90% RH and cannot stably exist under low ambient humidity conditions.

[0175] Example 7: p-Toluenesulfonate of Compound A Two solvent systems, methanol and acetone / water, were selected, and 1 equivalent of p-toluenesulfonic acid was added as raw material to prepare p-toluenesulfonic acid salt. Specific experimental information and results are shown in Table 19.

[0176] Characterization results showed that the ratio of p-toluenesulfonic acid to free base in the sample obtained using 1 equivalent of acid as a starting material was 1:1, resulting in the following two forms of p-toluenesulfonate: (1) Sample 1 obtained with methanol (p-toluenesulfonate Form DI of Compound A) is a monohydrate (see Figure 7-1 for XRPD characterization data and Table 20 below for characterization data), and (2) Sample 2 obtained with acetone / water (19 / 1) is a mixed crystal of a new crystalline form, Form 2 (Crystalline Form D-II), of p-toluenesulfonate of Compound A and the free base (see Figure 8-1 for XRPD characterization data).

[0177] [Table 19]

[0178] [Table 20]

[0179] The p-toluenesulfonate salt of Compound A, Form DI, has low crystallinity, irregular crystals, and small particles. The thermal analysis results are shown in Figure 7-2. The TGA pattern shows a weight loss of approximately 4.2% between room temperature and 150°C. The DSC pattern shows two broad endothermic peaks at 49°C and 167°C, the former of which is due to solvent loss and the latter of which may be a melting peak. NMR analysis revealed no residual organic solvent in the sample, indicating a 1:1 molar ratio of free base to p-toluenesulfonic acid. These results indicate that p-toluenesulfonate salt of Compound A, Form DI, is a monohydrate (theoretical water content of a monohydrate is 2.3%).

[0180] Characterization of Sample 2 of p-toluenesulfonate salt of Compound A revealed that the sample was highly crystalline and irregularly crystalline. However, as shown in the XRPD pattern (Figure 8-1), the sample contained a small amount of free base and was a mixed crystal of crystalline Form D-II and the free base. See Table 21 for characterization data. The TGA pattern showed a weight loss of approximately 3.8% between room temperature and 130°C, which is likely due to solvent loss. The DSC pattern showed three endothermic peaks at 70°C, 145°C, and 160°C, which are due to desolvation and melting, respectively (Figure 8-2). NMR results indicated that the sample contained approximately 0.1% residual acetone solvent, with a free base to acid ratio of 1:0.9. Therefore, crystalline Form D-II of p-toluenesulfonate salt of Compound A should be a monohydrate. Because the sample contained a small amount of free base, characterization data for pure crystalline Form D-II was not available.

[0181] [Table 21]

[0182] Example 8: Fumarate salt of Compound A Compound A obtained in Preparation 1 was dissolved in 20V of methanol, and 1 equivalent of fumaric acid was added to obtain a suspension. 12V of methanol solution was added, and the mixture was heated to 50°C and stirred for 3 hours. When the solution was still a suspension, the mixture was cooled to room temperature, stirred overnight, filtered, and dried at 50°C for 20 hours to obtain the fumarate salt of Compound A. This crystalline solid was defined as crystalline form E.

[0183] Form E of the fumarate salt of Compound A is an irregular crystal with a relatively low degree of crystallinity. See Figure 9-1 for XRPD characterization and Table 22 below for characterization data. Thermal analysis results are shown in Figure 9-2. The TGA pattern shows a weight loss of approximately 3.8% between room temperature and 150°C. The DSC pattern shows multiple thermal events, including a desolvation peak at 32°C, an exothermic crystalline transition peak at 159°C, and endothermic peaks at 222°C, 230°C, and 240°C, which are likely due to melting or decomposition. NMR revealed no residual organic solvent in the sample and a 1:1 ratio of fumaric acid to free base. Therefore, Form E of the fumarate salt is a monohydrate (theoretical water content of a monohydrate is 2.5%), but it has a low degree of crystallinity and dehydrates at a relatively low temperature.

[0184] [Table 22]

[0185] Example 9: Citrate salt of Compound A Compound A obtained in Preparation Example 1 was dissolved in 20V of methanol, and 1 equivalent of citric acid was added to obtain a suspension. 12V of methanol solution was added, and the mixture was stirred at 50°C for 3 hours. After the solution was still a suspension, it was cooled to room temperature, stirred at room temperature overnight, filtered, and dried at 50°C for 20 hours to obtain Sample 1.

[0186] Compound A obtained in Preparation Example 1 was added to 20V of acetone / water (4 / 1), heated to 50°C and dissolved, and 2 equivalents of citric acid were added to obtain a clear solution. After stirring for 1.5 hours, no solid precipitated. The solution was cooled to room temperature and stirred at room temperature overnight. After that, a solid precipitated. The solution was filtered and dried at 50°C for 6 hours to obtain Sample 2.

[0187] XRPD characterization (see Figure 10-1) indicates that Sample 1 and Sample 2 are identical, crystalline form F. See Table 23 below for characterization data. This crystalline form is irregular and has a relatively high degree of crystallinity. Thermal analysis results are shown in Figure 10-2. The TGA pattern shows a weight loss of approximately 1.6% between room temperature and 150°C. The DSC pattern shows three endothermic peaks at 30°C, 198°C, and 248°C, which are due to solvent loss, melting, and sample decomposition, respectively. H-NMR results indicate a 1:1 ratio of free base to citric acid in the sample. DVS measurements indicate that Form F absorbed approximately 3.8% moisture at 80% RH, and the crystalline form of the sample remained unchanged before and after DVS. The citrate salt may be a channel hydrate, and its water content changes with humidity.

[0188] [Table 23]

[0189] Example 10: Malate salt of Compound A Compound A obtained in Preparation Example 1 was taken, and 20V of methanol was added, and the mixture was stirred to dissolve. 1 equivalent of malic acid was added to obtain a suspension. 12V of methanol was added, and the mixture was heated to 50°C and stirred to obtain a clear solution. After 3 hours, the mixture was cooled to room temperature and stirred overnight. A solid precipitated out, which was filtered and dried at 50°C for 20 hours to obtain the malate salt of Compound A.

[0190] The resulting product was characterized by XRPD (see Figure 11-1) and identified as crystalline form G. The characterization data are shown in Table 24 below. Malate crystalline form G is highly crystalline, irregular, and aggregated. Thermal analysis (see Figure 11-2) showed that the TGA pattern showed a weight loss of approximately 4.2% between room temperature and 120°C, and the DSC pattern showed four endothermic peaks at 57°C, 175°C, 191°C, and 238°C, the former due to solvent loss, and the latter three due to melting and decomposition. NMR analysis showed that the ratio of malic acid to free base in the sample was 1:1, with no residual organic solvent. Therefore, the sample is a monohydrate (the theoretical water content of a monohydrate is 2.3%).

[0191] [Table 24]

[0192] Example 11: Succinate salt of Compound A Compound A obtained in Preparation Example 1 was added with 20V of acetone / water (9 / 1), heated to 50°C, and 1 equivalent of succinic acid was added. The solution was still a suspension, but some of the solids had dissolved. After stirring for 1 hour, the mixture was cooled to room temperature, 20V of ethyl acetate was added, and the mixture was stirred overnight. The mixture was filtered and dried at 50°C for 20 hours to obtain the succinate salt of compound A.

[0193] The RPD pattern showed a free base diffraction peak, NMR indicated that the molar ratio of free base to succinic acid in the sample was 3:2, and the DSC pattern showed a melting peak of the free base, suggesting that the free base and succinic acid did not react completely or that the succinate salt was prone to dissociation.

[0194] Example 12: Other salts of Compound A In the present invention, the reaction of the free base with other acids (sulfuric acid, methanesulfonic acid, tartaric acid, maleic acid, and phosphoric acid) was also attempted. Specific experimental procedures and results are shown in Table 25 below. The results showed that the free base of Compound A could not obtain crystalline samples by reacting with maleic acid and phosphoric acid, but could obtain a total of four crystalline samples by reacting with sulfuric acid, methanesulfonic acid, and tartaric acid. However, the obtained samples all had low crystallinity, and detailed characterization was not performed. Therefore, the free base could not obtain crystalline salts with acceptable crystallinity by reacting with sulfuric acid, methanesulfonic acid, tartaric acid, maleic acid, and phosphoric acid.

[0195] [Table 25]

[0196] Test Example 1: Free base crystal form conversion test of Compound A 1. Anhydrous crystal form conversion test 1) Preparation of initial mixed sample: Equal amounts of free base crystalline form AI and crystalline form A-IV samples of Compound A were taken, mixed uniformly, and sampled for XRPD characterization; 2) 1 mL of the corresponding solvent was added to the sample to form a suspension, which was then stirred at the corresponding temperature and sampled for XRPD characterization.

[0197] [Table 26]

[0198] 2. Competition experiments between anhydrous and solvated compounds and solvent activity experiments 1) Taking a mixed sample of equal amounts of different crystalline forms of the free base of Compound A, mixing them uniformly, and sampling for XRPD characterization; 2) 1 mL of the corresponding solvent was added to the mixed sample to form a suspension, which was stirred at room temperature and sampled for XRPD characterization.

[0199] [Table 27]

[0200] Test Example 2: Stability test Appropriate amounts of Compound A free base crystalline Form AI, Compound A hydrochloride monohydrate crystalline Form B, and Compound A benzenesulfonate crystalline Form C were opened and left for 7 days under three different conditions: 60°C, 92.5% RH, and 40°C / 75% RH. After that, HPLC and XRPD detection were performed on each of the 7-day samples to confirm their chemical and physical stability.

[0201] The results showed that Compound A free base crystalline form AI, Compound A hydrochloride monohydrate crystalline form B, and Compound A benzenesulfonate crystalline form C all maintained stable chemical properties and showed no obvious decomposition after 7 days at 60°C, 92.5% RH, and 40°C / 75% RH. XRPD results showed that Compound A free base crystalline form AI and Compound A hydrochloride monohydrate crystalline form B did not change crystal form after 7 days under the three conditions, while Compound A benzenesulfonate crystalline form C transformed into a new crystal form, possibly a hydrate, after 7 days at high humidity 92.5% RH, and did not change crystal form under the other two conditions.

[0202] In addition, in the present invention, the crystal form of the free base crystalline form AI of Compound A was examined by polishing or pulverizing, and it was confirmed that the crystal form of the sample did not change.

[0203] [Table 28]

[0204] Test Example 3: Comparison of the solubilities of Compound A free base crystalline form AI, Compound A hydrochloride monohydrate crystalline form B, and Compound A benzenesulfonate crystalline form C Compound A hydrochloride monohydrate crystalline Form B, Compound A benzenesulfonate crystalline Form C, and Compound A free base crystalline Form AI were each placed in water, SGF, FaSSIF, and FeSSIF to obtain 5 mg / mL sample solutions / suspensions, with two samples prepared in parallel for each medium. All samples were shaken at 200 rpm at 37°C for 24 hours, and appropriate amounts of the suspension were filtered at 0.5, 2, and 24 hours, respectively. The filtrate was analyzed by HPLC and pH was measured, and the filter cake was analyzed by XRPD.

[0205] The results showed that the free base and two salts had solubilities greater than 5 mg / mL in SGF. The free base had low solubility in water, approximately 0.003 mg / mL. Both salts had high solubility in water, with the benzenesulfonate salt having the highest solubility, greater than 5 mg / mL, at 24 hours. In FaSSIF and FeSSIF, the solubility of the hydrochloride salt was slightly higher than that of the free base. In FaSSIF, the solubility of the benzenesulfonate salt was lower than that of the free base and hydrochloride salt within 2 hours due to crystal form conversion.

[0206] XRPD measurements were performed on the filter cake and showed that during the solubility measurement process, the free base and benzenesulfonate salt became viscous in FeSSIF, and the benzenesulfonate salt became hydrochloride crystalline form I after 0.5 h in FeSSIF medium, while the XRPD patterns of the other remaining solids did not change.

[0207] From the above, the solubility of the free base crystalline form AI of Compound A in water was clearly lower than that of the two salts, and its solubility in the other three biologically relevant media was not significantly different from that of the two salts.

[0208] [Table 29]

[0209] Test Example 4: In vitro inhibition test of tumor cell proliferation by the free base crystalline form AI of Compound A 1. Test Method Tumor cell lines were cultured at 37°C in a 5% CO2 incubator. Periodically passaged cells were harvested during logarithmic growth phase and plated. Viable cells were counted by staining with trypan blue. The cell concentration was adjusted to the appropriate concentration. Cell suspension was added to each well of the culture plate, and cell-free culture medium was added to blank control wells. The culture plate was then cultured overnight in a 37°C, 5% CO2, 100% relative humidity incubator. Compound A (free base crystalline form AI) was serially diluted with DMSO from highest to lowest concentration (starting from 20 μM, diluted 1:3, giving eight concentration gradients, each with three replicate wells). The compound working solution was added to the cell culture plate, and the same final concentration of DMSO was added to the solvent control wells. The cells were returned to the incubator and cultured for 4 to 7 days. Cell viability was detected according to the instructions for the Promega CellTiter-Glo (CTG) Luminescent Cell Viability Detection Kit (Promega-G7573).

[0210] The inhibition rate (IR) of the test compound was calculated using the following formula: [Formula 1] IR(%)=[1-(RLU 化合物 -RLU ブランク対照 ) / (RLU 溶媒対照 -RLU ブランク対照 )] × 100%

[0211] Calculate the inhibition percentage of compounds at different concentrations in Excel, then use GraphPad Prism software to generate inhibition curve graphs and record the minimum inhibition percentage, maximum inhibition percentage, and IC 50 The relevant parameters were calculated, including IC, based on the following formula: 50 was calculated. [Formula 2] Y = minimum inhibition rate + (maximum inhibition rate - minimum inhibition rate) / (1 + 10^((LogIC 50 -X)*HillSlope)) X:log(concentration) Y: Response value, this value is negatively correlated with X HillSlope: Slope factor NOTE: If the maximum inhibition rate is <50%, the result is IC 50 > Expressed as maximum starting concentration.

[0212] 2. Test Results The results showed that Compound A had excellent inhibitory activity against the seven breast cancer cell lines tested, demonstrating clear superiority over the CDK4 / 6 selective inhibitor Palbociclib. Compound A exhibited superior inhibitory activity against the Palbociclib-resistant MDA-MB-468(Rb-) and Palbociclib-acquired resistant xMCF-7 / -Palbo-R. The superiority of Compound A treatment was particularly evident against the Palbociclib-resistant MDA-MB-468(Rb-), with a maximum inhibition rate exceeding 99%, compared with only 34% for Palbociclib. Compound A exhibited superior inhibitory activity to Palbociclib against HR+ / HER2- breast cancer cells T47D, triple-negative breast cancer cells HCC1187, and MDA-MB-231, and exhibited comparable inhibitory activity to Palbociclib against HR+ / HER2- breast cancer cells MCF-7. Compound A exhibited an IC50 of 1.20 for HR- / HER2+ breast cancer cells MDA-MB-453. 50 Values ​​are IC of Palbociclib 50 Although the dose of Compound A was approximately twice that of Palbociclib, the maximum inhibition rate for this cell line was nearly 100%, while Palbociclib only reached a maximum inhibition rate of approximately 74% even with increasing doses. Compound A also showed higher maximum inhibition rates for other breast cancer cell lines. This indicates that Compound A is superior to Palbociclib in terms of the maximum effect of inhibiting tumor cell proliferation.

[0213] As a direct downstream target of CDK4 / 6, Rb protein is considered to be the main biomarker for assessing CDK4 / 6 inhibitor sensitivity, and Rb protein loss (Rb-) indicates that the tumor is insensitive to CDK4 / 6 inhibitors (primary drug resistance). The results in Table 31 show that compound A has a significant inhibitory effect on Hep3B (Rb-) liver cancer cells, with a maximum inhibition rate of over 99%, suggesting that compound A has excellent therapeutic effect on liver cancer resistant to CDK4 / 6 inhibitors.

[0214] Compound A exhibits excellent inhibitory activity against Rb+ colon cancer cells COLO205, HT-29, and HCT116, Rb+ liver cancer cells Huh-7 and JHH-7, blood tumor cells MV-4-11 and MOLM-13, ovarian cancer cell A2780, small cell lung cancer cell NCI-H211, and malignant melanoma cell SK-MEL-5.

[0215] [Table 30]

[0216] [Table 31]

[0217] Test Example 5: Effect of particle size of free base crystalline form AI of Compound A on formulation dissolution The free base crystalline form AI of Compound A is a drug with low solubility, and the particle size of the raw material may affect the drug dissolution and content uniformity, so the particle size of the drug substance must be considered.

[0218] Using the free base crystalline form AI of Compound A as the raw material, raw materials with different particle sizes were selected, and common excipients such as fillers and disintegrants were added and pressed to produce tablets. The effect of the raw material particle size on the formulation dissolution was examined, and the results are shown in the table below.

[0219] [Table 32]

[0220] Test Example 6: Stability considerations of the free base crystalline form AI of Compound A The factors influencing the stability of the free base crystalline form AI of Compound A were investigated, and accelerated and long-term studies were completed, with the test results shown in the table below.

[0221] (1) Influence factor test

[0222] [Table 33]

[0223] Conclusion: The results of the investigation of influencing factors show that the properties of crystalline form AI of Compound A did not change significantly under the above test conditions. Under high temperature conditions, the related substances did not change significantly. Under high humidity conditions, the moisture absorption gain (weight increase due to moisture absorption) did not change significantly. The related substances did not change significantly compared to the 0 month condition. Under light irradiation conditions (illuminance 4500 lx ± 500 lx, total illuminance of the light source 1.2 × 10 6 lux·hr or more, and the energy of the near-ultraviolet lamp is 200W·hr / m 2 The crystal form AI of Compound A is relatively stable at the above temperature, and no obvious decomposition impurities were observed. This indicates that the crystal form AI of Compound A is excellent in stability.

[0224] (2) Accelerated testing Packaging: Imitation commercial packaging (medicinal low-density polyethylene bags used as inner packaging, polyester / aluminum / polyethylene pharmaceutical packaging composite bags used as outer packaging) Consideration conditions: 40℃±2℃ / 75%±5%RH

[0225] [Table 34]

[0226] (3) Long-term stability test Packaging: Imitation commercial packaging (medicinal low-density polyethylene bags used as inner packaging, polyester / aluminum / polyethylene pharmaceutical packaging composite bags used as outer packaging) Consideration conditions: 25℃±2℃ / 60%±5%RH

[0227] [Table 35]

[0228] Conclusion: After 6 months of storage under accelerated testing conditions (40°C ± 2°C / 75% ± 5% RH) and 18 months of storage under long-term stability testing conditions (25°C ± 2°C / 60% ± 5% RH), the sample's properties, loss on drying (weight loss due to drying), related substances, crystalline form, and content were essentially consistent with those at month 0, with no obvious tendency for increases. This indicates that crystalline form AI of Compound A has excellent stability.

Claims

1. A crystalline form of Compound A, whose structure is represented by the following formula (I), or a pharmaceutically acceptable salt thereof: 【Chemistry 1】

2. 2. The crystalline form of Compound A or a pharmaceutically acceptable salt thereof according to claim 1, wherein the DSC pattern of the crystalline form of Compound A has an endothermic peak at 256±3°C.

3. 3. The crystalline form of Compound A or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the crystalline form of Compound A is anhydrous.

4. The crystalline form of Compound A is crystalline form AI, and its X-ray powder diffraction pattern expressed as 2θ angles using Cu-Kα radiation has characteristic peaks at 5.2±0.2°, 11.4±0.2°, 15.7±0.2°, 20.2±0.2°, and 22.0±0.2°; Alternatively, the crystalline form of Compound A is crystalline form AI, and the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 5.2±0.2°, 9.5±0.2°, 11.4±0.2°, 15.7±0.2°, 16.3±0.2°, 20.2±0.2°, and 22.0±0.2°; Alternatively, the crystalline form of Compound A is crystalline form AI, and the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 5.2±0.2°, 9.5±0.2°, 10.4±0.2°, 11.4±0.2°, 12.8±0.2°, 14.1±0.2°, 15.7±0.2°, 16.3±0.2°, 18.6±0.2°, 20.2±0.2°, 22.0±0.2°, and 25.8±0.2°; Alternatively, the crystalline form of Compound A is crystalline form AI, and the X-ray powder diffraction pattern using Cu-Kα radiation is substantially as shown in FIG. 1-1. The crystalline form of Compound A according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

5. The crystalline form of Compound A is crystalline form AI, and the DSC pattern of crystalline form AI has an endothermic peak at 256±3°C; Alternatively, the DSC pattern of crystalline form AI is substantially as shown in Figures 1 to 3.

5. The crystalline form of Compound A according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof.

6. The pharmaceutically acceptable salt of the crystalline form of Compound A is selected from sulfate, methanesulfonate, tartrate, benzenesulfonate, hydrochloride, p-toluenesulfonate, fumarate, citrate, and malate, preferably benzenesulfonate, hydrochloride, p-toluenesulfonate, fumarate, citrate, and malate, more preferably hydrochloride and benzenesulfonate; Preferably, the hydrochloride salt is a hydrate, more preferably, the hydrochloride salt is hydrochloride monohydrate, and the molar ratio of Compound A, hydrochloric acid and water is 1:1:

1.

2. The crystalline form of Compound A according to claim 1, or a pharmaceutically acceptable salt thereof.

7. The crystalline form of the hydrochloride salt of Compound A is crystalline form B, and the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 4.1±0.2°, 8.9±0.2°, 14.3±0.2°, and 19.9±0.2°; Alternatively, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, and the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 4.1±0.2°, 8.9±0.2°, 12.3±0.2°, 14.3±0.2°, 15.4±0.2°, 16.8±0.2°, and 19.9±0.2°; Alternatively, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, and the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 4.1±0.2°, 8.3±0.2°, 8.9±0.2°, 9.5±0.2°, 12.3±0.2°, 14.3±0.2°, 15.0±0.2°, 15.4±0.2°, 16.8±0.2°, 19.1±0.2°, 19.9±0.2°, 20.6±0.2°, and 25.5±0.2°; Alternatively, the crystalline form of the hydrochloride salt of Compound A is crystalline form B, and the X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 4.1±0.2°, 8.3±0.2°, 8.9±0.2°, 9.5±0.2°, 11.1±0.2°, 12.3±0.2°, 14.3±0.2°, 15.0±0.2°, 15.4±0.2°, 16.8±0.2°, 17.4±0.2°, 19.1±0.2°, 19.9±0.2°, 20.6±0.2°, 21.6±0.2°, 25.2±0.2°, 25.5±0.2°, 26.6±0.2°, and 29.1±0.2°; Alternatively, the crystalline form of Compound A hydrochloride is crystalline form B, and the X-ray powder diffraction pattern using Cu-Kα radiation is substantially as shown in Figure 5-1.

7. The crystalline form of Compound A according to claim 6, or a pharmaceutically acceptable salt thereof.

8. 8. The crystalline form of Compound A or a pharmaceutically acceptable salt thereof according to claim 6 or 7, wherein the crystalline form of Compound A hydrochloride is crystalline form B, and the weight loss in the range of room temperature to 130°C in a TGA pattern is about 4.4%.

9. 9. The crystalline form of Compound A or a pharmaceutically acceptable salt thereof according to any one of claims 6 to 8, wherein the crystalline form of Compound A hydrochloride is crystalline form B, and the DSC pattern has an endothermic peak at 253°C, or the DSC pattern has two endothermic peaks at 27°C and 253°C.

10. 7. The crystalline form of Compound A or a pharmaceutically acceptable salt thereof according to claim 6, wherein the pharmaceutically acceptable salt of the crystalline form of Compound A is selected from benzenesulfonate salts, preferably the benzenesulfonate salts are anhydrous, and preferably the molar ratio of Compound A free base to benzenesulfonic acid in the benzenesulfonate salt is 1:

1.

11. The crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and its X-ray powder diffraction pattern expressed as 2θ angles using Cu-Kα radiation has characteristic peaks at 8.5±0.2°, 15.3±0.2°, 19.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°; Alternatively, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and its X-ray powder diffraction pattern expressed as 2θ angles using Cu-Kα radiation has characteristic peaks at 8.5±0.2°, 15.3±0.2°, 16.4±0.2°, 17.1±0.2°, 17.8±0.2°, 19.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°; Alternatively, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and its X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 8.5±0.2°, 15.3±0.2°, 16.4±0.2°, 17.1±0.2°, 17.8±0.2°, 18.1±0.2°, 19.2±0.2°, 20.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°; Alternatively, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and its X-ray powder diffraction pattern expressed in 2θ angles using Cu-Kα radiation has characteristic peaks at 8.5±0.2°, 11.7±0.2°, 12.7±0.2°, 13.3±0.2°, 14.2±0.2°, 15.3±0.2°, 16.4±0.2°, 17.1±0.2°, 17.8±0.2°, 18.1±0.2°, 19.2±0.2°, 19.5±0.2°, 20.2±0.2°, 21.7±0.2°, 22.1±0.2°, and 26.1±0.2°; Alternatively, the crystalline form of benzenesulfonate salt of Compound A is crystalline form C, and the X-ray powder diffraction pattern using Cu-Kα radiation is substantially as shown in FIG. 6-1.

11. The crystalline form of Compound A according to claim 10, or a pharmaceutically acceptable salt thereof.

12. The crystalline benzenesulfonate salt of Compound A has an endothermic peak at 235±3°C in a DSC pattern; Alternatively, the crystalline form of benzenesulfonate salt of Compound A has a DSC pattern substantially as shown in Figure 6-2.

12. The crystalline form of Compound A according to claim 10 or 11, or a pharmaceutically acceptable salt thereof.

13. 13. A pharmaceutical composition comprising Compound A in crystalline form according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, and optionally one or more pharmaceutically acceptable excipients. Pharmaceutical compositions.

14. Use of the crystalline form of Compound A or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, or the pharmaceutical composition according to claim 13, in the manufacture of an antitumor drug, comprising: Preferably, the tumor is selected from a drug-sensitive tumor or a drug-resistant tumor; Preferably, the sensitive or drug-resistant tumor is selected from a solid tumor or a blood tumor, and preferably, the solid tumor is selected from fibrosarcoma, salivary gland cancer, liver cancer, colon cancer, bladder cancer, pharyngeal cancer, breast cancer, prostate cancer, glioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, pancreatic cancer, skin cancer, lymphoma, gastric cancer, multiple myeloma, brain tumor, lung cancer, and malignant melanoma, and the blood tumor is selected from acute myeloid leukemia; More preferably, the sensitive or drug-resistant tumor is selected from breast cancer, lung cancer, colon cancer, liver cancer, ovarian cancer, malignant melanoma, and acute myeloid leukemia; More preferably, the breast cancer is selected from HR+ / HER2- breast cancer, triple-negative breast cancer, and HR- / HER2+ breast cancer, the acute myeloid leukemia is selected from myelomonocytic leukemia, and the lung cancer is selected from non-small cell lung cancer. use.

15. The tumor is selected from a drug-resistant tumor, and the drug-resistant tumor is selected from a primary drug-resistant tumor or an acquired drug-resistant tumor; Preferably, the drug-resistant tumor is selected from a CDK4 / 6 inhibitor-resistant tumor, and further selected from a CDK4 / 6 inhibitor-primary resistant tumor or a CDK4 / 6 inhibitor-acquired resistant tumor, and the CDK4 / 6 inhibitor is selected from palbociclib, abemaciclib, ribociclib, and dalpiciclib, preferably palbociclib; More preferably, the CDK4 / 6 inhibitor-resistant tumor is selected from breast cancer and liver cancer, even more preferably from palbociclib-resistant breast cancer and liver cancer, and even more preferably from palbociclib-resistant breast cancer or liver cancer, or palbociclib-acquired-resistant breast cancer or liver cancer; More preferably, the palbociclib primary resistant breast cancer or palbociclib acquired resistant breast cancer is selected from palbociclib primary resistant or acquired resistant HR+ / HER2- breast cancer, triple negative breast cancer, and HR- / HER2+ breast cancer.

15. The use according to claim 14.

16. 1. Use of the crystalline form of Compound A or a pharmaceutically acceptable salt thereof in the manufacture of an antitumor medicament in combination with another clinically / pharmaceutically acceptable drug, comprising: the other drug is selected from endocrine therapy drugs, such as anastrozole, letrozole, exemestane, tamoxifen, toremifene, fulvestrant, megestrol, fluoxymesterone, ethinylestradiol, preferably fulvestrant; Preferably, the other drug and the crystalline form of Compound A or a pharmaceutically acceptable salt thereof may be contained in the same dosage unit, or may be contained in different dosage units and prepared in a combined package form, for example, a kit product.

16. Use according to claim 14 or 15.

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