Crystalline form of cyclin-dependent kinase inhibitor, pharmaceutical composition, and use thereof

Abstract

The present disclosure relates to a crystalline form of a cyclin-dependent kinase inhibitor, a pharmaceutical composition, and use thereof. Specifically, the structure of the cyclin-dependent kinase inhibitor is represented by formula I.

Description

Crystallographic form, pharmaceutical composition and uses of a cyclin-dependent kinase inhibitor Technical Field

[0001] This disclosure relates to a crystalline form, pharmaceutical composition, and use of a cyclin-dependent kinase inhibitor. Background Technology

[0002] Cyclin-dependent kinases (CDKs) are an important class of kinases and play a crucial role in the division and proliferation of cancer cells and the transcriptional regulation of oncogenes. Currently, more than 20 subtypes of cyclin-dependent kinases (CDKs) have been discovered. Due to the sequence and structural similarity of the kinase domains among CDK family members, selective and precise regulation of each subtype is a significant challenge.

[0003] In recent years, the biggest advance in the field of breast cancer treatment has undoubtedly been the use of CDK4 / 6 alone or in combination with endocrine therapy for hormone receptor-positive advanced breast cancer. For example, palbociclib, ribociclib, and abemaciclib have been approved in combination with aromatase inhibitors for the treatment of hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer in postmenopausal women. Palbociclib and abemaciclib have also been approved in combination with fulvestrant for the treatment of hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer in postmenopausal women after disease progression following endocrine therapy (Nature Reviews (2016) 13:417-430, J Clin Oncol 2017, 35, 2875-2884). Although CDK4 / 6 inhibitors have shown significant clinical efficacy in estrogen receptor ER-positive metastatic breast cancer, like other kinases, their effects may be limited over time by the development of primary or acquired resistance.

[0004] In the cell cycle pathway, the phosphorylation level of Rb protein is jointly regulated by CDK4 / 6 and CDK2. In individuals resistant to CDK4 / 6 inhibitors, Rb protein dysfunction, increased CCNE1 expression, and increased MYC protein levels have been observed. Cyclin E is overexpressed in various cancers, particularly breast cancer, lung cancer, leukemia, and lymphoma (Guo Cuiping et al., Regulation of Cyclin E and Malignant Tumors. International Journal of Oncology, 2012, 39(005):337-340). Cyclin E amplification or overexpression is also associated with poor prognosis in ovarian cancer, gastric cancer, endometrial cancer, and other cancers.

[0005] Studies have shown that inhibiting CDK2 kinase induces the activity of tumor cells with high CCNE1 expression, but inhibiting CDK2 activity alone is insufficient to maintain tumor suppression. Research has found that breast cancer cells treated with CDK2 inhibitors activate cyclin A2 through the CDK4 pathway, thereby promoting upregulation of CDK2 protein expression and leading to resistance to CDK2 inhibitors (Cell. 2023 Jun 8; 186(12):2628-2643.e21.). Therefore, simultaneously inhibiting the activity of CDK4 and CDK2 is of great significance for long-term tumor suppression. Furthermore, simultaneous inhibition of CDK4 and CDK2 may play an important role in alleviating resistance to CDK4 / 6 inhibitors, and even CDK2 inhibitor resistance.

[0006] PCT / CN2024 / 107786 discloses a new class of CDK inhibitors, the structure of which is shown below: The crystal form of a pharmaceutically active ingredient often affects its chemical stability. Different crystallization and storage conditions can lead to changes in the crystal structure of the compound, and sometimes even result in other crystal forms. Therefore, it is essential to conduct in-depth research on the crystal forms of the aforementioned compounds to improve their various properties. Summary of the Invention

[0007] This disclosure provides the crystal form of the compound shown in formula (I). Specifically, crystal forms A, B, C, D, E, F, G, and H can be selected.

[0008] This disclosure provides a crystal form A of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 10.116, 10.666, 16.013, 21.453, and 24.060.

[0009] In some embodiments, the crystal form A of the compound of formula (I) provided in this disclosure has characteristic peaks at 5.259, 6.335, 8.108, 10.116, 10.666, 16.013, 21.453, 24.060, 27.547, and 28.363 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0010] In some embodiments, the X-ray powder diffraction pattern of the compound of formula (I) provided in this disclosure, expressed as a diffraction angle 2θ, has characteristic peaks at 5.259, 6.335, 8.108, 10.116, 10.666, 11.819, 12.837, 13.984, 15.458, 16.013, 17.261, 18.563, 19.601, 20.318, 20.854, 21.453, 22.242, 24.060, 25.691, 26.951, 27.547, 27.906, 28.363, 29.730, and 32.464.

[0011] In some embodiments, the X-ray powder diffraction pattern of the crystal form A of the compound of formula (I) provided in this disclosure, expressed in terms of the diffraction angle 2θ, is shown in Figure 1.

[0012] This disclosure provides a crystal form B of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 10.016, 10.424, 14.522, 14.933, and 19.468.

[0013] In some embodiments, the crystal form B of the compound represented by formula (I) provided in this disclosure is characterized by having characteristic peaks at 6.032, 7.640, 10.016, 10.424, 13.413, 14.522, 14.933, 19.468, 20.307, 21.807, and 23.377 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0014] In some embodiments, the crystal form B of the compound represented by formula (I) provided in this disclosure is characterized by having characteristic peaks at 6.032, 7.640, 10.016, 10.424, 13.413, 14.522, 14.933, 17.105, 18.218, 19.468, 20.307, 21.807, 23.377, 24.083, 24.862, 27.100, 28.577, 30.393, and 31.915 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0015] In some embodiments, the X-ray powder diffraction pattern of crystal form B of the compound of formula (I) provided in this disclosure, expressed in terms of diffraction angle 2θ, is shown in Figure 2.

[0016] This disclosure provides a crystal form C of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 15.090, 15.613, 20.258, 20.873, and 23.934.

[0017] In some embodiments, the crystal form C of the compound represented by formula (I) provided in this disclosure is characterized by having characteristic peaks at 10.164, 10.795, 15.090, 15.613, 20.258, 20.873, 23.934, 27.906, and 28.092 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0018] In some embodiments, the crystal form C of the compound represented by formula (I) provided in this disclosure is characterized by having characteristic peaks at 10.164, 10.795, 15.090, 15.613, 17.142, 18.181, 19.443, 20.258, 20.873, 22.227, 23.934, 24.677, 25.567, 27.164, 27.906, 28.092, 29.502, and 30.615 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0019] In some embodiments, the X-ray powder diffraction pattern of the crystal form C of the compound of formula (I) provided in this disclosure, expressed in terms of the diffraction angle 2θ, is shown in Figure 3.

[0020] This disclosure provides a crystal form D of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 5.649, 10.118, 11.350, 17.179, and 23.027.

[0021] In some embodiments, the crystal form D of the compound represented by formula (I) provided in this disclosure is characterized by having characteristic peaks at 5.649, 10.118, 11.350, 12.255, 14.061, 17.179, 21.376, 23.027, 24.049, and 25.973 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0022] In some embodiments, the crystal form D of the compound represented by formula (I) provided in this disclosure is characterized by having characteristic peaks at 5.649, 6.549, 8.630, 10.118, 11.350, 12.255, 14.061, 17.179, 18.764, 19.258, 21.376, 22.190, 23.027, 24.049, 25.973, 26.616, 27.869, and 28.834 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0023] In some embodiments, the X-ray powder diffraction pattern of the crystal form D of the compound of formula (I) provided in this disclosure, expressed in terms of the diffraction angle 2θ, is shown in Figure 4.

[0024] This disclosure provides a crystal form E of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 6.539, 14.106, 16.786, and 19.183.

[0025] In some embodiments, the crystal form E of the compound of formula (I) provided in this disclosure has characteristic peaks at 4.299, 6.539, 7.009, 11.314, 12.279, 14.106, 16.786, and 19.183 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0026] In some embodiments, the crystal form E of the compound of formula (I) provided in this disclosure has characteristic peaks at 4.299, 6.539, 7.009, 11.314, 12.279, 14.106, 16.786, 19.183, 21.893, 22.301, 23.340, and 29.242 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0027] In some embodiments, the X-ray powder diffraction pattern of the crystal form E of the compound of formula (I) provided in this disclosure, expressed in terms of the diffraction angle 2θ, is shown in Figure 5.

[0028] This disclosure provides a crystal form F of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 14.311, 17.127, 20.124, and 23.034.

[0029] In some embodiments, the crystal form F of the compound of formula (I) provided in this disclosure has characteristic peaks at 5.487, 9.941, 10.906, 11.537, 14.311, 17.127, 20.124, and 23.034 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0030] In some embodiments, the crystal form F of the compound of formula (I) provided in this disclosure has characteristic peaks at 5.487, 8.531, 9.941, 10.906, 11.537, 13.133, 14.311, 17.127, 18.070, 20.124, 23.034, and 26.124 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0031] In some embodiments, the X-ray powder diffraction pattern of the crystal form F of the compound of formula (I) provided in this disclosure, expressed in terms of the diffraction angle 2θ, is shown in Figure 6.

[0032] This disclosure provides a crystal form G of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 5.240, 15.833, and 21.209.

[0033] In some embodiments, the crystal form G of the compound of formula (I) provided in this disclosure has characteristic peaks at 5.240, 10.536, 15.833, 20.510, 21.209, and 25.736 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0034] In some embodiments, the crystal form G of the compound of formula (I) provided in this disclosure has characteristic peaks at 5.240, 6.361, 8.065, 10.062, 10.536, 15.833, 20.510, 21.209, 22.098, 23.868, 24.215, 25.736, 28.108, 29.542, 31.036, 32.026, and 37.603 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0035] In some embodiments, the X-ray powder diffraction pattern of the crystal form G of the compound of formula (I) provided in this disclosure, expressed in terms of the diffraction angle 2θ, is shown in Figure 7.

[0036] This disclosure provides a crystal form H of the compound shown in formula (I), characterized in that the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 10.079, 10.595, 13.699, 15.718, and 20.963.

[0037] In some embodiments, the crystal form H of the compound of formula (I) provided in this disclosure has characteristic peaks at 5.235, 6.294, 10.079, 10.595, 13.699, 15.718, 17.066, 20.963, and 23.495 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0038] In some embodiments, the X-ray powder diffraction pattern of the compound of formula (I) provided in this disclosure, expressed as a diffraction angle 2θ, has characteristic peaks at 5.235, 6.294, 8.177, 10.079, 10.595, 13.699, 15.718, 17.066, 20.963, 23.495, 23.832, 25.106, 26.977, and 27.661.

[0039] In some embodiments, the X-ray powder diffraction pattern of the crystal form H of the compound of formula (I) provided in this disclosure, expressed in terms of the diffraction angle 2θ, is shown in Figure 8.

[0040] In some embodiments, the crystal form of the compound represented by formula (I) provided in this disclosure has a 2θ angle error range of ±0.20.

[0041] This disclosure provides a method for preparing crystal form A of the compound shown in formula (I) above, comprising the following steps:

[0042] The compound shown in formula (I) is mixed with solvent I, stirred to crystallize, and then separated into solid and liquid components. Solvent I is selected from one or more of water, ester solvents, ketone solvents, ether solvents, and hydrocarbon solvents.

[0043] In an optional embodiment, the method for preparing crystal form A of the compound represented by formula (I) provided in this disclosure uses ethyl acetate or isopropyl acetate as the ester solvent, 2-butanone or methyl isobutyl ketone as the ketone solvent, tetrahydrofuran as the ether solvent, and n-heptane or cyclohexane as the hydrocarbon solvent.

[0044] This disclosure provides a method for preparing crystal form B of the compound shown in formula (I) as described above, comprising the following steps: dissolving the compound shown in formula (I) in an ether solvent, evaporating and crystallizing, and separating the solid and liquid phases.

[0045] In an optional embodiment, the method for preparing crystal form B of the compound represented by formula (I) provided in this disclosure uses 1,4-dioxane as the ether solvent.

[0046] This disclosure provides a method for preparing crystal form C of the compound shown in formula (I) above, selected from the following methods:

[0047] (i) Dissolve the compound shown in formula (I) in an ether solvent, volatilize and crystallize, and then separate the solid and liquid phases;

[0048] (ii) Mix the compound shown in formula (I) with a halohydrocarbon solvent, stir to precipitate crystals, and then separate the solid and liquid components.

[0049] (iii) Dissolve the compound shown in formula (I) in solvent II to form solution 1, mix solution 1 with solvent III, stir to precipitate crystals, and separate the solid and liquid. Solvent II is selected from ether solvents, and solvent III is selected from one or more of water, ether solvents, and hydrocarbon solvents.

[0050] In an optional embodiment, the method for preparing crystal form C of the compound represented by formula (I) provided in this disclosure is selected from the following methods:

[0051] (i) Dissolve the compound shown in formula (I) in an ether solvent, volatilize and crystallize, and separate the solid and liquid phases. The ether solvent is propylene glycol methyl ether.

[0052] (ii) The compound shown in formula (I) is mixed with a halohydrocarbon solvent, stirred to crystallize, and then separated into solid and liquid phases. The halohydrocarbon solvent is chloroform.

[0053] (iii) Dissolve the compound shown in formula (I) in solvent II to form solution 1, mix solution 1 with solvent III, stir to precipitate crystals, and separate the solid and liquid. Solvent II is selected from ether solvents, and solvent III is selected from one or more of water, ether solvents, and hydrocarbon solvents. The ether solvent in solvent II is selected from tetrahydrofuran, 1,4-dioxane or a mixture thereof; the ether solvent in solvent III is methyl tert-butyl ether, and the hydrocarbon solvent is n-heptane.

[0054] This disclosure provides a method for preparing crystal form D of the compound represented by formula (I) as described above, selected from the following methods:

[0055] (i) Dissolve the compound shown in formula (I) in solvent IV to form solution 1, mix solution 1 with solvent V, stir to precipitate crystals, and separate solid and liquid. Solvent IV is selected from ketone solvents, mixed solvents of ketone solvent and water, or halogenated hydrocarbon solvents. Solvent V is selected from water, hydrocarbon solvents, or ether solvents.

[0056] (ii) Mix the compound shown in formula (I) with solvent VI, stir to crystallize, and separate the solid and liquid. Solvent VI is selected from ketone solvents or halogenated hydrocarbon solvents.

[0057] In an optional embodiment, the method for preparing crystal form D of the compound represented by formula (I) provided in this disclosure is selected from the following methods:

[0058] (i) The compound shown in formula (I) is dissolved in solvent IV to form solution 1. Solution 1 is mixed with solvent V, stirred to induce crystallization, and then the solid and liquid are separated. Solvent IV is selected from ketone solvents, mixed solvents of ketone solvent and water, or halogenated hydrocarbon solvents. Solvent V is selected from water, hydrocarbon solvents, or ether solvents. The ketone solvent in solvent IV is acetone, and the halogenated hydrocarbon solvent is dichloromethane. The hydrocarbon solvent in solvent V is n-heptane, and the ether solvent is methyl tert-butyl ether.

[0059] (ii) Mix the compound shown in formula (I) with solvent VI, stir to precipitate crystals, and separate the solid and liquid. Solvent VI is selected from ketone solvents or halogenated hydrocarbon solvents. The ketone solvent in solvent VI is acetone, and the halogenated hydrocarbon solvent is dichloromethane.

[0060] This disclosure provides a method for preparing crystal form E of the compound shown in formula (I) as described above, comprising the following steps: mixing the compound shown in formula (I) with an ether solvent, stirring to precipitate crystals, and separating the solid and liquid phases; optionally, the ether solvent is methyl tert-butyl ether.

[0061] This disclosure provides a method for preparing crystal form F of the compound shown in formula (I) above, comprising the following steps: mixing the compound shown in formula (I) with an ether solvent, stirring to precipitate crystals, and separating solids and liquids. Optionally, the ether solvent is propylene glycol methyl ether.

[0062] This disclosure provides a method for preparing crystal form G of the compound represented by formula (I) as described above, which is selected from the following methods:

[0063] (i) Mix the compound shown in formula (I) with solvent VII, stir to precipitate crystals, and separate the solid and liquid components. Solvent VII is selected from water, alcohol solvents, ketone solvents, halogenated hydrocarbon solvents, ester solvents and mixed solvents of hydrocarbons.

[0064] (ii) Mix the compound shown in formula (I) with solvent VIII, volatilize and crystallize, and separate the solid and liquid. The solvent VIII is selected from a mixture of ketone solvents, halogenated hydrocarbon solvents, nitrile solvents, water and alcohol solvents.

[0065] (iii) Dissolve the compound shown in formula (I) in solvent IX to form solution 1, and mix solution 1 with solvent X, wherein solvent IX is selected from alcohol solvents, ether solvents, ketone solvents, halogenated hydrocarbon solvents or mixtures thereof, and solvent X is selected from hydrocarbon solvents, water, and ether solvents.

[0066] In an optional embodiment, the method for preparing the crystal form G of the compound represented by formula (I) provided in this disclosure is selected from the following methods:

[0067] (i) The compound shown in formula (I) is mixed with solvent VII, stirred to crystallize, and then separated into solid and liquid components. Solvent VII is selected from a mixture of alcohol solvents, ketone solvents, halogenated hydrocarbon solvents, ester solvents, and hydrocarbon solvents. The alcohol solvent is isopropanol, the ester solvent is ethyl acetate, the ketone solvent is acetone, the halogenated hydrocarbon solvent is selected from dichloromethane or trichloromethane, and the hydrocarbon solvent is n-heptane.

[0068] (ii) The compound shown in formula (I) is mixed with solvent VIII, and the mixture is volatilized and crystallized, and then separated into solid and liquid components. Solvent VIII is selected from a mixture of ketone solvents, halogenated hydrocarbon solvents, nitrile solvents, water and alcohol solvents. The ketone solvent is acetone, the halogenated hydrocarbon solvent is selected from dichloromethane or trichloromethane, the nitrile solvent is acetonitrile, and the alcohol solvent is isopropanol.

[0069] (iii) Dissolve the compound shown in formula (I) in solvent IX to form solution 1, and mix solution 1 with solvent X. Solvent IX is selected from alcohol solvents, ether solvents, ketone solvents, halogenated hydrocarbon solvents or mixtures thereof. Solvent X is selected from hydrocarbon solvents, water, and ether solvents. The alcohol solvent in solvent IX is ethanol, the ether solvent is tetrahydrofuran, the ketone solvent is acetone, the halogenated hydrocarbon solvent is dichloromethane, the hydrocarbon solvent in solvent X is selected from n-heptane, and the ether solvent is selected from methyl tert-butyl ether.

[0070] This disclosure provides a method for preparing crystal form H of the compound shown in formula (I) above, comprising the following steps: mixing the compound shown in formula (I) with an ester solvent, evaporating and crystallizing, and separating the solid and liquid phases; optionally, the ester solvent is ethyl acetate.

[0071] In some embodiments, the "ester solvent" described in this disclosure is selected from ethyl acetate, triethyl glycerol, methyl acetate, n-butyl acetate, ethyl propionate, ethyl butyrate, ethyl lactate, triethyl phosphate, ethyl hexanoate, and ethyl formate.

[0072] In some embodiments, the "alcohol solvent" described in this disclosure is selected from methanol, ethanol, n-propanol, isopropanol, or mixtures thereof.

[0073] In some embodiments, the "ketone solvent" described in this disclosure is selected from acetone, methyl ethyl ketone, methyl propyl ketone, methyl isopropyl ketone, or mixtures thereof.

[0074] In some embodiments, the "nitrile solvent" described in this disclosure is selected from acetonitrile, propionitrile, or mixtures thereof.

[0075] In some embodiments, the "hydrocarbon solvent" described in this disclosure is selected from nitromethane, n-heptane, n-pentane, cyclohexane, toluene, p-xylene, or mixtures thereof.

[0076] In some embodiments, the "ether solvent" described in this disclosure is selected from methyl tert-butyl ether, propylene glycol methyl ether, isopropyl ether, tetrahydrofuran, dioxane, or mixtures thereof.

[0077] In some embodiments, the "halogenated hydrocarbon solvent" described in this disclosure is selected from dichloromethane or trichloromethane.

[0078] This disclosure also provides a pharmaceutical composition comprising a crystal form of the compound of formula (I) above, specifically selected from crystal forms A, B, C, D, E, F, G, H, or mixtures thereof, and at least one pharmaceutically acceptable excipient.

[0079] This disclosure also provides a pharmaceutical composition prepared from a crystal form of the compound represented by formula (I) above, specifically selected from crystal forms A, B, C, D, E, F, G, H, or a mixture thereof, and at least one pharmaceutically acceptable excipient.

[0080] This disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing a crystal form of a compound of formula (I), specifically selected from crystal forms A, B, C, D, E, F, G, H, or a mixture thereof, with at least one pharmaceutically acceptable excipient.

[0081] In some embodiments, the pharmaceutical composition contains 0.01% to 99.99% pharmaceutically acceptable excipients based on the total weight of the composition; in some embodiments, the pharmaceutical composition contains 0.1% to 99.9% pharmaceutically acceptable excipients; in some embodiments, the pharmaceutical composition contains 0.5% to 99.5% pharmaceutically acceptable excipients; in some embodiments, the pharmaceutical composition contains 1% to 99% pharmaceutically acceptable excipients; and in some embodiments, the pharmaceutical composition contains 2% to 98% pharmaceutically acceptable excipients.

[0082] In some embodiments, the pharmaceutically acceptable excipient may be, for example, a carrier, transporter, diluent, and / or delivery polymer.

[0083] This disclosure also provides crystal forms of the compounds represented by the aforementioned formula (I), specifically selected from crystal forms A, B, C, D, E, F, G, H, or mixtures thereof, or the use of the aforementioned pharmaceutical compositions in the preparation of medicaments for treating or preventing diseases or conditions associated with abnormal activity of serine / threonine kinases.

[0084] This disclosure also provides crystal forms of the compounds represented by the aforementioned formula (I), specifically selected from crystal forms A, B, C, D, E, F, G, H, or mixtures thereof, or the use of the aforementioned pharmaceutical compositions in the preparation of medicaments for treating or preventing diseases or conditions associated with abnormal activity of CDK2.

[0085] This disclosure also provides crystal forms of the compounds represented by the aforementioned formula (I), specifically selected from crystal forms A, B, C, D, E, F, G, H, or mixtures thereof, or the use of the aforementioned pharmaceutical compositions in the preparation of medicaments for treating or preventing diseases or conditions associated with abnormal activity of CDK4.

[0086] This disclosure also provides crystal forms of the compounds represented by the aforementioned formula (I), specifically selected from crystal forms A, B, C, D, E, F, G, H, or mixtures thereof, or the use of the aforementioned pharmaceutical compositions in the preparation of medicaments for treating or preventing diseases or conditions, said diseases or conditions being selected from proliferative diseases, inflammatory diseases, autoinflammatory diseases, autoimmune diseases, or infectious diseases.

[0087] In some implementations, the disease or condition is a proliferative disease.

[0088] In some implementations, the proliferative disease is selected from breast cancer (e.g., triple-negative breast cancer or ER-negative, PR-negative and Her2-positive breast cancer or CDK4 / 6 inhibitor-resistant breast cancer), colorectal cancer, lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, prostate cancer, Ewing's sarcoma, osteoma, neuroblastoma, cervical cancer, ovarian cancer, gastric cancer, and liver cancer.

[0089] This disclosure, in another aspect, discloses a compound.

[0090] The "2θ or 2θ angle" mentioned in this disclosure refers to the diffraction angle, where θ is the Bragg angle, and the unit is ° or degree; the error range of 2θ for each characteristic peak is ±0.20 (including the case where the number has more than one decimal place after rounding), specifically -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

[0091] The numerical values ​​in this disclosure, such as those relating to the content of certain substances, are calculated data and inevitably contain a certain degree of error. Generally, ±10% is within the reasonable error range. The error may vary to some extent depending on the context in which it is used, but this variation shall not exceed ±10%, and may be ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%, preferably ±5%.

[0092] The starting material used in the crystal form preparation method disclosed herein can be any form of compound, including but not limited to: amorphous, arbitrary crystal form, hydrate, solvate, etc.

[0093] The drying temperature described in this disclosure is generally 25℃-100℃, preferably 40℃-70℃, and can be dried under normal pressure or reduced pressure.

[0094] The crystallization methods described in this disclosure include room temperature crystallization, cooling crystallization, solvent evaporation crystallization, and seed crystallization induction. The cooling temperature is selected from below 65°C, preferably from -10°C to 60°C. Stirring can also be performed during the crystallization process.

[0095] The “differential scanning calorimetry or DSC” described in this disclosure refers to measuring the temperature difference and heat flow difference between the sample and the reference material during the sample heating or isothermal process, in order to characterize all physical and chemical changes related to thermal effects and obtain phase transition information of the sample.

[0096] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "Guiding Principles on Hygroscopicity of Drugs" in Part IV of the 2020 edition of the Chinese Pharmacopoeia,

[0097] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0098] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15%;

[0099] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15% but not less than 2%;

[0100] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0101] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0102] The “excipients” described in this disclosure include, but are not limited to, any adjuvants, carriers, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, or emulsifiers that have been approved by the U.S. Food and Drug Administration for use in humans or livestock. Attached Figure Description

[0103] Figure 1 shows the XRPD spectrum of crystal form A of the compound represented by formula (I).

[0104] Figure 2 shows the XRPD spectrum of crystal form B of the compound shown in formula (I).

[0105] Figure 3 shows the XRPD spectrum of crystal form C of the compound shown in formula (I).

[0106] Figure 4 shows the XRPD spectrum of crystal form D of the compound shown in formula (I).

[0107] Figure 5 shows the XRPD spectrum of crystal form E of the compound shown in formula (I).

[0108] Figure 6 shows the XRPD spectrum of the compound F represented by formula (I).

[0109] Figure 7 shows the XRPD spectrum of the compound G represented by formula (I).

[0110] Figure 8 shows the XRPD spectrum of the compound H represented by formula (I).

[0111] Figure 9 shows the amorphous XRPD spectrum of the compound represented by formula (I). Detailed Implementation

[0112] The present disclosure is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the present disclosure.

[0113] Experimental methods in the embodiments of this disclosure that do not specify specific conditions are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents whose specific source is not specified are commercially available conventional reagents.

[0114] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (LCMS). NMR shifts (δ) were expressed in 10⁻¹⁰. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard. The spatial configurations of the optical isomers (isomers) of the compounds were further confirmed by measuring single-crystal parameters.

[0115] HPLC determinations were performed using a Waters ACQUITY ultra high performance LC, Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200LC high performance liquid chromatograph (ACQUITY UPLC BEH C18 1.7UM 2.1*50MM column, Ultimate XB-C18 3.0*150mm column, or Ultimate C18 2.1*30mm column).

[0116] MS measurements were performed using a Waters SQD2 mass spectrometer in positive / negative ion mode, with a mass scan range of 100–1200.

[0117] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15 mm to 0.2 mm, and the diameter of the silica gel plates used for thin-layer chromatography separation and purification products is 0.4 mm to 0.5 mm.

[0118] Rapid column purification systems use either the Combiflash Rf150 (TELEDYNE ISCO) or Isolara One (Biotage).

[0119] For normal column chromatography, Yantai Huanghai silica gel (100-200 mesh, 200-300 mesh, or 300-400 mesh) is generally used as the support, or Changzhou Santai pre-filled ultrapure normal phase silica gel column (40-63 μm) is used. 12g, 25g, 40g, 80g or other sizes).

[0120] Reversed-phase column chromatography typically uses Changzhou Sante pre-packed ultrapure C18 silica gel columns (20-45μm). 40g, 80g, 120g, 220g or other sizes).

[0121] The high-pressure column purification system uses Waters AutoP, in conjunction with the Waters XBridge BEH C18 OBD Prep Column. 5μm, 19mm x 150mm or Atlantis T3 OBD Prep Column, 5μm, 19mm x 150mm.

[0122] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as Shanghai Titan Technology, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0123] Unless otherwise specified in the examples, all reactions can be carried out under a nitrogen atmosphere.

[0124] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0125] A hydrogen atmosphere refers to a reaction flask connected to a hydrogen balloon with a volume of approximately 1L.

[0126] The pressurized hydrogenation reaction was performed using a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0127] The hydrogenation reaction is usually carried out under vacuum, filled with hydrogen gas, and repeated 3 times.

[0128] The microwave reaction was performed using a CEM Discover-S 908860 microwave reactor.

[0129] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0130] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃~30℃.

[0131] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compounds, and the developing solvent system for TLC included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate system, C: petroleum ether / ethyl acetate system, D: petroleum ether / ethyl acetate / methanol, and E: petroleum ether / tetrahydrofuran system. The volume ratio of the solvent was adjusted according to the polarity of the compounds, and small amounts of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0132] The purification process in the examples can be performed using a silica gel column for forward column chromatography. The eluent system includes, but is not limited to: A: dichloromethane / methanol system, B: petroleum ether / ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound. A small amount of basic or acidic reagents such as triethylamine or acetic acid can also be added for adjustment.

[0133] The purification process in the examples uses a C18 reversed-phase chromatography column. The eluent system includes, but is not limited to, acetonitrile / water system (the solvent ratio is adjusted according to the polarity of the actual compound). Small amounts of alkaline or acidic reagents such as ammonium formate, ammonia, or formic acid can also be added for adjustment.

[0134] The solvents used in the purification process of the examples include, but are not limited to, dichloromethane, methanol, petroleum ether, ethyl acetate, ethanol, acetonitrile, and water, and the slurry is slurried using a single solvent or a combination of multiple solvents.

[0135] Example 1. Preparation of (S)-4-(4-((5-chloro-4-(8-fluoro-2-(2-hydroxypropyl-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazanephrine-6-yl)pyrimidin-2-amino)-3-fluorophenyl)morpholin-3-one (1)

[0136] Step 1: 4-(3-fluoro-4-nitrophenyl)morpholin-3-one (1a)

[0137] 1bb, i.e., 4-bromo-2-fluoro-1-nitrobenzene (5 g, 22.7 mmol), was added to dioxane (40 mL). Then, 1aa, morpholino-3-one (2.76 g, 27.3 mmol), cesium carbonate (14.8 g, 45.4 mmol), and XantPhos Pd G3 (645 mg, 0.68 mmol) were added at room temperature. The mixture was purged three times with nitrogen and reacted at 110 °C. The reaction solution was concentrated, and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 1:1) to give compound 1a (1.82 g, yield 33.4%).

[0138] Step 2: 4-(4-amino-3-fluorophenyl)morpholin-3-one (1b)

[0139] Compound 1a (1.82 g, 7.58 mmol) was added to ethanol (12 mL) and water (3 mL), along with iron powder (1.70 g, 30.3 mmol) and ammonium chloride (1.62 g, 30.3 mmol). The mixture was reacted at 80 °C and filtered while hot. The filtrate was poured into water (50 mL), extracted with ethyl acetate (15 mL × 2), the organic phases were combined, dried, filtered, and pulped (petroleum ether / ethyl acetate = 3:1). After filtration, compound 1b (1.15 g, 72.3% yield) was obtained.

[0140] MS(ESI)m / z = 210.2[M+H] + .

[0141] Step 3: (S)-4-(4-((5-chloro-4-(8-fluoro-2-(2-hydroxypropyl-2-yl)-3-methyl-3,4-dihydro-5-oxa-1,2a-diazanaphth-6-yl)pyrimidin-2-amino)-3-fluorophenyl)morpholin-3-one (1)

[0142] Compound 1b (32 mg, 0.15 mmol) was added to dioxane (4 mL), followed by compound A10 (50 mg, 0.12 mmol, synthesized and resolved according to patent WO2022166799), cesium carbonate (78 g, 0.24 mmol), and XantPhos Pd G3 (9 mg, 0.01 mmol) at room temperature. The mixture was purged with nitrogen and reacted in a microwave oven at 110 °C for 1 hour. The reaction solution was filtered, concentrated, and the residue was purified by C18 reversed-phase chromatography (acetonitrile:water = 4:1) to give compound 1 (21.6 mg, yield 30.0%).

[0143] MS(ESI)m / z = 571.2[M+H] + .

[0144] 1H NMR (400MHz, DMSO-d6) δ9.57(s,1H),8.55(s,1H),7.72(t,1H),7.39(dd,1H),7.25–7.17(m,1H),6.99(d,1H),5.82(s,1H),5. 26-5.22(m,1H),4.50(d,1H),4.28–4.18(m,3H),3.97(dd,2H),3.75(dd,2H),1.67(s,3H),1.62(s,3H),1.46(d,J=6.6Hz,3H).

[0145] Example 2. Preparation of free crystal form A

[0146] Weigh approximately 10 mg of the compound shown in formula (I), add 1.0 mL of deionized water, stir to precipitate crystals, centrifuge, and dry the solid under vacuum. The product was identified by X-ray powder diffraction and defined as crystal form A. The XRPD spectrum is shown in Figure 1, and the positions of its characteristic peaks are shown in Table 1.

[0147] The DSC spectrum shows that the endothermic peaks are 135.86℃, 158.77℃, and 264.15℃.

[0148] The TGA spectrum showed a weight loss of 0.71% at 31℃-94℃, 6.21% at 95℃-171℃, and 8.32% at 212℃-313℃.

[0149] Table 1. Positions of characteristic peaks of free-state crystal form A

[0150] Example 3. Preparation of free crystal form A

[0151] Weigh approximately 100 mg of the compound shown in formula (I), add 1.0 mL of ethyl acetate, stir to induce crystallization, centrifuge, and dry the solid under vacuum. X-ray powder diffraction analysis confirmed that it was crystal form A.

[0152] Example 4. Preparation of free crystal form A

[0153] Weigh approximately 10 mg of the compound shown in formula (I), add 1.0 mL of a solvent selected from Table 2, stir to induce crystallization, centrifuge, and then dry the solid under vacuum. X-ray powder diffraction analysis confirmed that the solid was crystal form A.

[0154] Table 2. Solvent Systems

[0155] Example 5. Preparation of free crystal form B

[0156] Weigh approximately 10 mg of the compound shown in formula (I), add 0.15 mL of 1,4-dioxane, and allow it to evaporate and crystallize at room temperature to obtain a solid. X-ray powder diffraction analysis was performed, and the product was defined as crystal form B. The XRPD spectrum is shown in Figure 2, and the positions of its characteristic peaks are shown in Table 3.

[0157] The DSC spectrum shows that the endothermic peak values ​​are 120.81℃, 155.84℃, 161.30℃, 178.80℃, and 267.27℃.

[0158] The TGA spectrum showed a weight loss of 0.50% at 30℃-76℃, 9.46% at 76℃-191℃, and 10.25% at 191℃-319℃.

[0159] Table 3. Positions of characteristic peaks of free-state B crystal form

[0160] Example 6. Preparation of free crystalline form C

[0161] Weigh approximately 10 mg of the compound shown in formula (I), add 0.25 mL of propylene glycol methyl ether, volatilize and crystallize at room temperature, and identify the product by X-ray powder diffraction. The product is defined as crystal form C. The XRPD spectrum is shown in Figure 3, and the positions of its characteristic peaks are shown in Table 4.

[0162] The DSC spectrum shows endothermic peaks at 109.49℃, 142.83℃, 167.49℃, and 255.65℃. The TGA spectrum shows a weight loss of 10.85% between 30℃ and 208℃, and a weight loss of 6.98% between 209℃ and 322℃.

[0163] Table 4. Positions of characteristic peaks of C in free-state crystal forms

[0164] Example 7. Preparation of free crystalline form C

[0165] Weigh approximately 10 mg of the compound shown in formula (I), add 0.25 mL of chloroform, stir to induce crystallization, and X-ray powder diffraction analysis confirmed that it is crystal form C.

[0166] Example 8. Preparation of free crystalline form C

[0167] Weigh approximately 10 mg of the compound shown in formula (I), dissolve it in 0.20 mL of tetrahydrofuran, add 1.0 mL of a solvent selected from Table 5, stir to induce crystallization, and X-ray powder diffraction analysis showed that it was crystal form C.

[0168] Table 5. Solvent Systems

[0169] Example 9. Preparation of free crystalline form C

[0170] Weigh approximately 10 mg of the compound shown in formula (I), dissolve it in 0.15 mL of 1,4-dioxane, add 1.0 mL of n-heptane, stir to induce crystallization, and X-ray powder diffraction analysis confirmed that it is crystal form C.

[0171] Example 10. Preparation of free crystal form D

[0172] Weigh approximately 120 mg of the compound shown in formula (I), dissolve it in 2.0 mL of 10% water / acetone (v / v), add 5.1 mL of deionized water, stir to induce crystallization, filter, and dry the solid under vacuum to obtain the title product.

[0173] X-ray powder diffraction analysis determined the product to be crystal form D. The XRPD spectrum is shown in Figure 4, and the positions of its characteristic peaks are shown in Table 6.

[0174] The DSC spectrum shows endothermic peaks at 62.83℃, 164.97℃, and 261.30℃. The TGA spectrum shows a weight loss of 0.92% between 32℃ and 94℃.

[0175] DVS testing showed that under normal storage conditions (i.e., 25°C, 60% RH), the sample's moisture absorption weight gain was approximately 0.92%; under accelerated testing conditions (i.e., 70% RH), the moisture absorption weight gain was approximately 1.02%; and under extreme conditions (90% RH), the moisture absorption weight gain was approximately 1.24%. Furthermore, retesting of the crystal form after DVS testing showed no change in crystal form.

[0176] Table 6. Positions of characteristic peaks of free-state crystal form D

[0177] Example 11. Preparation of free crystal form D

[0178] Weigh about 10 mg of the compound shown in formula (I), dissolve it in 0.8 mL of acetone, add 2.0 mL of a solvent selected from Table 7, stir to crystallize, centrifuge and then vacuum dry the solid to obtain a solid, which was identified as crystal form D by X-ray powder diffraction.

[0179] Table 7. Solvent Systems

[0180] Example 12. Preparation of free crystal form D

[0181] Weigh about 10 mg of the compound shown in formula (I), dissolve it in 0.25 mL of dichloromethane, add 2.0 mL of methyl tert-butyl ether, stir to crystallize, centrifuge and dry the solid under vacuum to obtain a solid, which is identified as crystal form D by X-ray powder diffraction.

[0182] Example 13. Preparation of free crystal form D

[0183] Weigh about 10 mg of the compound shown in formula (I), add 0.1 mL of a solvent selected from Table 8, stir to crystallize, centrifuge and dry the solid under vacuum to obtain the title product.

[0184] Table 8. Solvent Systems

[0185] Example 14. Preparation of free crystal form E

[0186] Weigh approximately 10 mg of the compound shown in formula (I), add 1.0 mL of methyl tert-butyl ether, stir to crystallize, centrifuge and vacuum dry the solid. The product is identified by X-ray powder diffraction and is defined as crystal form E. The XRPD spectrum is shown in Figure 5, and the positions of its characteristic peaks are shown in Table 9.

[0187] The DSC spectrum shows that the endothermic peaks are 144.12℃ and 266.77℃.

[0188] The TGA spectrum showed a weight loss of 0.35% at 30℃-103℃ and a weight loss of 9.00% at 103℃-191℃.

[0189] Table 9. Positions of E characteristic peaks in free-state crystal forms

[0190] Example 15. Preparation of free crystal form F

[0191] Weigh approximately 10 mg of the compound shown in formula (I), add 0.1 mL of propylene glycol methyl ether, stir to crystallize, centrifuge and vacuum dry the solid. The product is identified by X-ray powder diffraction and is defined as crystal form F. The XRPD spectrum is shown in Figure 6, and the positions of its characteristic peaks are shown in Table 10.

[0192] The DSC spectrum shows an endothermic peak at 165.87℃. The TGA spectrum shows a weight loss of 1.58% between 37℃ and 187℃.

[0193] Table 10. Positions of characteristic F peaks in free-state crystal forms

[0194] Example 16. Preparation of free crystal form G

[0195] Weigh approximately 10 mg of the compound shown in formula (I), add 0.2 mL of isopropanol, stir to crystallize, centrifuge, and dry the solid under vacuum. The product is identified by X-ray powder diffraction and is defined as crystal form G. The XRPD spectrum is shown in Figure 7, and the positions of its characteristic peaks are shown in Table 11.

[0196] The DSC spectrum shows that the endothermic peaks are 159.44℃ and 263.83℃.

[0197] The TGA spectrum showed a weight loss of 0.16% at 32℃-102℃, 5.92% at 102℃-183℃, and 7.08% at 219℃-303℃.

[0198] Table 11. Positions of characteristic peaks of G in free-state crystal forms

[0199] Example 17. Preparation of free crystal form G

[0200] Weigh approximately 10 mg of the compound shown in formula (I), add 0.2 mL of ethyl acetate / n-heptane (1:1), stir to induce crystallization, centrifuge, and then dry the solid under vacuum. X-ray powder diffraction analysis confirmed that the crystal form is G.

[0201] Example 18. Preparation of free crystal form G

[0202] Weigh approximately 10 mg of the compound shown in formula (I), add 1.0 mL of a solvent selected from Table 12, allow it to evaporate and crystallize at room temperature, and X-ray powder diffraction analysis shows that it is crystal form G.

[0203] Table 12. Solvent Systems

[0204] Example 19. Preparation of free crystal form G

[0205] Weigh approximately 10 mg of the compound shown in formula (I), add 0.2 mL of acetonitrile, and allow it to evaporate and crystallize at room temperature. X-ray powder diffraction analysis confirmed that the crystal form is G.

[0206] Example 20. Preparation of free crystal form G

[0207] Weigh about 10 mg of the compound shown in formula (I), dissolve it in 0.4 mL of ethanol, add 2.0 mL of n-heptane, stir to induce crystallization, centrifuge and dry the solid under vacuum. X-ray powder diffraction analysis showed that it was crystal form G.

[0208] Example 21. Preparation of free crystal form G

[0209] Weigh about 10 mg of the compound shown in formula (I), dissolve it in 0.20 mL of tetrahydrofuran, add 1.0 mL of a solvent selected from Table 13, stir to crystallize, centrifuge and dry the solid under vacuum. X-ray powder diffraction analysis showed that it was crystal form G.

[0210] Table 13. Solvent Systems

[0211] Example 22. Preparation of free crystal form G

[0212] Weigh about 10 mg of the compound shown in formula (I), dissolve it in 0.8 mL of acetone, add 2.0 mL of a solvent selected from Table 13, stir to induce crystallization, and X-ray powder diffraction analysis showed that it was crystal form G.

[0213] Table 14. Solvent Systems

[0214] Example 23. Preparation of free crystal form G

[0215] Weigh about 10 mg of the compound shown in formula (I), dissolve it in 0.25 mL of dichloromethane, add 2.0 mL of methyl tert-butyl ether, stir to induce crystallization, and X-ray powder diffraction analysis shows that it is crystal form G.

[0216] Example 24. Preparation of free crystal form G

[0217] Weigh approximately 10 mg of the compound shown in formula (I), add 0.1 mL of acetone or dichloromethane, stir to induce crystallization, and X-ray powder diffraction analysis shows that it is crystal form G.

[0218] Example 25. Preparation of free crystalline form H

[0219] Weigh about 10 mg of the compound shown in formula (I), add 0.4 mL of ethyl acetate, volatilize and crystallize at room temperature, and identify the product by X-ray powder diffraction. The product is defined as crystal form H. The XRPD spectrum is shown in Figure 8, and the positions of its characteristic peaks are shown in Table 15.

[0220] Table 15. Positions of characteristic peaks of H in free-state crystal form

[0221] Example 26. Preparation of free amorphous form

[0222] Weigh approximately 10 mg of the compound shown in formula (I), add 0.2 mL of methanol, and allow it to evaporate at room temperature to precipitate a solid.

[0223] X-ray powder diffraction analysis showed that it was in a free, amorphous state, as shown in Figure 9.

[0224] Example 27. Preparation of free amorphous form

[0225] Weigh approximately 10 mg of the compound shown in formula (I), add 0.2 mL of a solvent selected from Table 16, and allow it to evaporate at room temperature to precipitate a solid. X-ray powder diffraction analysis showed that the solid was in a free, amorphous state.

[0226] Table 16. Solvent Systems

[0227] Example 28. Preparation of free amorphous form

[0228] Weigh about 10 mg of the compound shown in formula (I), dissolve it in 0.2 mL of methanol, add 1.0 mL of a solvent selected from Table 17, stir to precipitate, and X-ray powder diffraction analysis shows that it is a free amorphous state.

[0229] Table 17. Solvent Systems

[0230] Example 29. Stability of Influencing Factors

[0231] The D-type crystals were laid flat in the open, and the stability of the samples was investigated under light (4500 Lux), high temperature (40℃, 60℃), and high humidity (RH 75%, RH 92.5%) conditions. The sampling period was one month.

[0232] Table 18. Factors affecting the stability of D crystal form

[0233] Conclusion: Crystal form D has good physical stability under the influence of various factors. Its chemical purity decreases slightly under light irradiation, but its chemical stability is good under high temperature and high humidity conditions.

[0234] Experimental Example 30. Long-term / accelerated stability

[0235] The D-type closed crystal was subjected to stability tests under conditions of 25℃ / 60%RH and 40℃ / 75%RH, with a sampling period of 6 months.

[0236] Table 19. Long-term / accelerated stability of D crystal form

[0237] Conclusion: Crystal form D exhibits good physical and chemical stability after 6 months of long-term / accelerated storage.

[0238] Biological tests

[0239] The present disclosure is further described and explained below with reference to test examples, but these embodiments are not intended to limit the scope of the present disclosure.

[0240] Test Example 1: Inhibitory activity test of the disclosed compound against ovarian cancer cells (OVCAR3)

[0241] Table 20. Experimental Materials and Instruments

[0242] 1) Experimental Procedure

[0243] Ovarian cancer cells (OVCAR3) were cultured in RPMI 1640 medium with 10% FBS in a 37% CO2 incubator. On day one, cells were plated in 384-well plates at a concentration of 800 cells / well and cultured overnight. On day two, cells were treated with compounds at a maximum concentration of 10 μM, diluted 3-fold, resulting in 9 concentrations, with a final concentration of 0.1% DMSO. After 7 days of continued culture, cell viability was assessed using the CTG Cell Viability Detection Reagent (MCE), following the instructions provided with the kit. Data were processed and IC50 was calculated using GraphPad Prism 8. 50 The calculation formula is Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope)).

[0244] X: Logarithm of compound concentration; Y: Inhibition percentage.

[0245] Table 21. IC50 of the compounds disclosed herein against OVCAR3 50 (nM)

[0246] Test Example 2: Inhibitory activity test of the disclosed compound against triple-negative breast cancer cells (HCC1806)

[0247] 1) Experimental materials and instruments

[0248] Table 22. Experimental Materials and Instruments

[0249] 2) Experimental Procedure

[0250] Triple-negative breast cancer cells HCC1806 (Nanjing Kober Biotechnology Co., Ltd.) were cultured in RPMI 1640 medium with 10% FBS at 37°C in a cell culture incubator with 5% CO2. On day one, cells were seeded in 384-well plates at a concentration of 200 cells / well and cultured overnight. On day two, compound treatments were performed at a maximum concentration of 10 μM, with 3-fold dilutions, resulting in 9 concentrations, and a final DMSO concentration of 0.2%. After 7 days of continued culture, cell viability was assessed using the CTG Cell Viability Detection Reagent (MCE), following the instructions provided in the kit. Data were processed using XLfit, and IC50 was calculated. 50 The calculation formula is Y = Bottom + (Top - Bottom) / (1 + 10^(LogIC)). 50-X)*HillSlope)).

[0251] X: Logarithm of compound concentration; Y: Inhibition percentage.

[0252] Table 23. IC50 of the compounds disclosed herein against HCC1806 50 (nM) Note: PF-07220060 (WO2019207463A Example A94) and PF-07104091 (WO2020157652A Example13) are Pfizer's selective CDK4 inhibitor and CDK2 inhibitor, respectively.

[0253] In experiments with this cell line, Example 1 showed better inhibitory activity compared to the clinical molecular CDK4 inhibitor PF-07220060 and the CDK2 inhibitor PF-07104091.

[0254] Test Example 3: Detection of the activity of the disclosed compounds against cyclin-dependent kinases (CDK2 / CycE1, CDK4 / CycD1)

[0255] 1) Experimental materials and instruments

[0256] Table 24. Experimental Materials and Instruments

[0257] 2) Experimental Procedure

[0258] Add the analyte compound at a maximum concentration of 1 μM using an Echo 650, performing a 3-fold serial dilution for a total of 9 concentration points. Seal the assay plate and centrifuge at 1000g for 1 minute. Prepare the 2.5× enzyme in 1× kinase buffer (50 mM HEPES, pH 7.5, 0.0015% Brij-35, 1 M DTT). Add 10 μl of the 2.5× enzyme to the 384-well assay plate, centrifuge at 1000g for 30 seconds, and incubate at room temperature for 10 minutes. Prepare the 2.5× substrate and ATP mixture in 1× kinase buffer, add 10 μl of the 2.5× substrate and ATP mixture to start the reaction. Centrifuge the plate at 1000g for 30 seconds, seal the assay plate, and incubate at room temperature for 1 hour. Add 25 μL of stop solution reagent (100 mM HEPES, pH 7.5, 0.015% Brij-35, 0.2% Coating Reagent #3, 50 mM EDTA).

[0259] Read conversion rate data from CaliperEZ Reader. Copy conversion rate data from CaliperEZ Reader. Convert conversion rate to inhibition rate data. Inhibition % = (max - conversion) / (max - min) * 100.

[0260] “min” represents the reading of the control wells without enzyme; “max” represents the reading of the control wells with DMSO added.

[0261] Fitting ICs using XLFit excel add-in version 5.4.0.8 50 Value, Fitting formula: Y = Bottom + (Top - Bottom) / (1 + (IC) 50 / X)^HillSlope).

[0262] Table 25. IC50 values ​​of the compounds disclosed herein for CDK2 / CycE1 and CDK4 / CycD1 50 value Note: Example 1 of the patent based on CN116217588A is used as Comparative Example 1 of this application.

[0263] Compound 1 exhibited good inhibitory activity against both CDK4 / CycD1 and CDK2 / CycE1. Compared to Comparative Example 1, Example 1 showed stronger inhibitory activity against CDK2 / CycE1, and therefore potentially alleviated some CDK4 / 6 inhibitor resistance issues. Example 1 may also have a wider range of indications.

[0264] Test Example 4. Inhibitory activity test of the disclosed compound against human breast cancer cells (MDA-MB-231)

[0265] Table 26. Experimental Materials and Instruments

[0266] Experimental steps

[0267] Human breast cancer cells MDA-MB-231 were cultured in DMEM medium containing 10% FBS at 37°C and 5% CO2. On day one, cells were seeded in 384-well plates at a concentration of 600 cells / well and cultured overnight. On day two, compound treatments were performed at a maximum concentration of 10 μM, with 3-fold dilutions, resulting in 9 concentrations, and a final DMSO concentration of 0.2%. After 7 days of continued culture, cell viability was assessed using the CTG Cell Viability Detection Reagent, following the instructions provided in the kit. Data were processed using XLFit, and IC50 was calculated. 50The calculation formula is Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope)).

[0268] X: Logarithm of compound concentration; Y: % inhibition.

[0269] Table 27. IC50 of the compounds of this disclosure against MDA-MB-231 50 (nM)

[0270] In experiments with this cell line, Example 1 showed better inhibitory activity compared to the clinical molecular CDK4 inhibitor PF-07220060 and the CDK2 inhibitor PF-07104091.

[0271] Test Example 5. Inhibitory activity test of the disclosed compound against human non-small cell lung cancer cells (A549).

[0272] Table 28. Experimental Materials and Instruments

[0273] Experimental steps

[0274] Human non-small cell lung cancer cells (A549) were cultured in F12K medium (10% FBS) at 37°C in a 5% CO2 incubator. On day one, cells were seeded in 384-well plates at a concentration of 300 cells / well and cultured overnight. On day two, compound treatments were performed at a maximum concentration of 10 μM, with 3-fold dilutions, resulting in 9 concentrations, and a final DMSO concentration of 0.2%. After 5 days of continued culture, cell viability was assessed using the CTG Cell Viability Detection Reagent, following the instructions provided with the kit. Data were processed using XLFit, and IC50 was calculated. 50 The calculation formula is Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope)).

[0275] X: Logarithm of compound concentration; Y: % inhibition.

[0276] Table 29. IC50 of the compounds disclosed herein against A549 50 (nM)

[0277] In experiments with this cell line, Example 1 showed better inhibitory activity compared to the clinical molecular CDK4 inhibitor PF-07220060 and the CDK2 inhibitor PF-07104091.

[0278] Test Example 6. Inhibitory Activity Test of the Compounds Disclosed Against Human Prostate Cancer Cells (22RV1)

[0279] Table 30. Experimental Materials and Instruments

[0280] Experimental steps

[0281] Human prostate cancer cells 22RV1 were cultured in RPMI 1640 medium with 10% FBS at 37°C and 5% CO2 in a cell culture incubator. On day one, cells were plated in 384-well plates at a concentration of 500 cells / well and cultured overnight. On day two, cells were treated with compounds at a maximum concentration of 10 μM, diluted 3-fold, resulting in 9 concentrations, with a final DMSO concentration of 0.2%. After 5 days of continued culture, cell viability was assessed using the CTG Cell Viability Detection Reagent, following the instructions provided in the kit. Data were processed using XLFit, and IC50 was calculated. 50 The calculation formula is Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope)).

[0282] X: Logarithm of compound concentration; Y: % inhibition.

[0283] Table 31. IC50 of the compounds of this disclosure against 22RV1 50 (nM)

[0284] In experiments with this cell line, Example 1 showed better inhibitory activity compared to the clinical molecular CDK2 inhibitor PF-07104091.

[0285] Test Example 7. Inhibitory Activity Test of the Compounds Disclosed Against Human Breast Cancer Cells (SUM-149PT)

[0286] Table 32. Experimental Materials and Instruments

[0287] Experimental steps

[0288] Human breast cancer cells SUM-149PT were cultured in DMEM medium containing 10% FBS at 37°C in a 5% CO2 incubator. On day one, cells were seeded in 384-well plates at a concentration of 800 cells / well and cultured overnight. On day two, compound treatments were performed at a maximum concentration of 10 μM, with 3-fold dilutions, resulting in 9 concentrations, and a final DMSO concentration of 0.2%. After 5 days of continued culture, cell viability was assessed using the CTG Cell Viability Detection Reagent, following the instructions provided in the kit. Data were processed using XLFit, and IC50 was calculated. 50 .

[0289] The calculation formula is Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope)).

[0290] X: Logarithm of compound concentration; Y: % inhibition.

[0291] Table 33. IC50 of the compounds disclosed herein against SUM-149PT 50 (nM)

[0292] In experiments with this cell line, Example 1 showed better inhibitory activity compared to the clinical molecular CDK2 inhibitor PF-07104091.

[0293] Test Example 8. Inhibitory activity test of the disclosed compounds against CDK4 / 6 inhibitor-resistant human breast cancer cells (MCF-7-pebocillin-resistant cell line).

[0294] 1) Experimental materials and instruments

[0295] Table 34. Experimental Materials and Instruments

[0296] 2) Experimental Procedure

[0297] MCF-7 / Palbo-R cell source: MCF7 cells (Nanjing Kober Biotechnology Co., Ltd.) were treated with 1 μM palbociclib for a long period of time to induce drug resistance.

[0298] Palbociclib-resistant breast cancer cell line MCF-7 / Palbo-R was cultured in DMEM containing 10% FBS in a 37% CO2 incubator. On day 1, cells were seeded in 384-well plates at a concentration of 800 cells / well and cultured overnight. On day 2, the cells were treated with the compound at a maximum concentration of 10 μM, diluted 3-fold, resulting in 9 concentrations, with a final DMSO concentration of 0.2%. After 5 days of continued culture, cell viability was assessed using a CellTiter-Glo luminescence assay kit, following the instructions provided with the kit. Data were processed using XLfit, and IC50 was calculated. 50 .

[0299] The inhibition rate (IR) of the detected compound is calculated using the following formula: IR (%) = (1 – (RLU compound – RLU blank control) / (RLU solvent control – RLU blank control)) * 100%.

[0300] Table 35. IC50 of the disclosed compounds against MCF-7 / Palbo-R 50 (nM)

[0301] In experiments using this cell line, the molecule from Example 1 exhibited better inhibitory activity compared to the clinical molecules CDK4 inhibitor PF-07220060 and CDK2 inhibitor PF-07104091.

Claims

1. A crystal form of the compound shown in formula (I), said crystal form being selected from crystal forms A, B, C, D, E, F, G, and H, wherein crystal form A is characterized in that, X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ. Characteristic peaks are observed at 10.116, 10.666, 16.013, 21.453, and 24.

060. The crystal form B is characterized by having characteristic peaks at 10.016, 10.424, 14.522, 14.933, and 19.468 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form C is characterized by having characteristic peaks at 15.090, 15.613, 20.258, 20.873, and 23.934 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form D is characterized by having characteristic peaks at 5.649, 10.118, 11.350, 17.179, and 23.027 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form E is characterized in that the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 6.539, 14.106, 16.786, and 19.

183. The crystal form F is characterized by having characteristic peaks at 14.311, 17.127, 20.124, and 23.034 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form G is characterized by having characteristic peaks at 5.240, 15.833, and 21.209 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form H is characterized in that the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 10.079, 10.595, 13.699, 15.718, and 20.

963.

2. The crystal form of the compound of formula (I) according to claim 1, wherein crystal form A is characterized in that, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 5.259, 6.335, 8.108, 10.116, 10.666, 16.013, 21.453, 24.060, 27.547, and 28.

363. The crystal form B is characterized by having characteristic peaks at 6.032, 7.640, 10.016, 10.424, 13.413, 14.522, 14.933, 19.468, 20.307, 21.807, and 23.377 in its X-ray powder diffraction pattern expressed as a diffraction angle of 2θ. The crystal form C is characterized by having characteristic peaks at 10.164, 10.795, 15.090, 15.613, 20.258, 20.873, 23.934, 27.906, and 28.092 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form D is characterized by having characteristic peaks at 5.649, 10.118, 11.350, 12.255, 14.061, 17.179, 21.376, 23.027, 24.049, and 25.973 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form E is characterized in that the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 4.299, 6.539, 7.009, 11.314, 12.279, 14.106, 16.786, and 19.

183. The crystal form F is characterized in that the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 5.487, 9.941, 10.906, 11.537, 14.311, 17.127, 20.124, and 23.

034. The crystal form G is characterized in that the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 5.240, 10.536, 15.833, 20.510, 21.209, and 25.

736. The crystal form H is characterized by having characteristic peaks at 5.235, 6.294, 10.079, 10.595, 13.699, 15.718, 17.066, 20.963, and 23.495 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

3. The crystal form of the compound of formula (I) according to claim 1, wherein crystal form A is characterized in that, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 5.259, 6.335, 8.108, 10.116, 10.666, 11.819, 12.837, 13.984, 15.458, 16.013, 17.261, 18.563, 19.601, 20.318, 20.854, 21.453, 22.242, 24.060, 25.691, 26.951, 27.547, 27.906, 28.363, 29.730, and 32.

464. The crystal form B is characterized by having characteristic peaks at 6.032, 7.640, 10.016, 10.424, 13.413, 14.522, 14.933, 17.105, 18.218, 19.468, 20.307, 21.807, 23.377, 24.083, 24.862, 27.100, 28.577, 30.393, and 31.915 in its X-ray powder diffraction pattern expressed as a diffraction angle of 2θ. The crystal form C is characterized by having characteristic peaks at 10.164, 10.795, 15.090, 15.613, 17.142, 18.181, 19.443, 20.258, 20.873, 22.227, 23.934, 24.677, 25.567, 27.164, 27.906, 28.092, 29.502, and 30.615 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form D is characterized by having characteristic peaks at 5.649, 6.549, 8.630, 10.118, 11.350, 12.255, 14.061, 17.179, 18.764, 19.258, 21.376, 22.190, 23.027, 24.049, 25.973, 26.616, 27.869, and 28.834 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form E is characterized by having characteristic peaks at 4.299, 6.539, 7.009, 11.314, 12.279, 14.106, 16.786, 19.183, 21.893, 22.301, 23.340, and 29.242 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form F is characterized by having characteristic peaks at 5.487, 8.531, 9.941, 10.906, 11.537, 13.133, 14.311, 17.127, 18.070, 20.124, 23.034, and 26.124 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form G is characterized by having characteristic peaks at 5.240, 6.361, 8.065, 10.062, 10.536, 15.833, 20.510, 21.209, 22.098, 23.868, 24.215, 25.736, 28.108, 29.542, 31.036, 32.026, and 37.603 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ. The crystal form H is characterized by having characteristic peaks at 5.235, 6.294, 8.177, 10.079, 10.595, 13.699, 15.718, 17.066, 20.963, 23.495, 23.832, 25.106, 26.977, and 27.661 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

4. The crystal form of the compound of formula (I) according to claim 1, wherein crystal form A is characterized in that, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 1. The crystal form B is characterized in that the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 2; The crystal form C is characterized by the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ, as shown in Figure 3. The crystal form D is characterized in that the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 4; The crystal form E is characterized in that the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 5. The crystal form F is characterized in that the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 6; The crystal form G is characterized in that the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 7. The crystal form H is characterized by an X-ray powder diffraction pattern expressed in terms of a diffraction angle of 2θ, as shown in Figure 8.

5. The crystal form of the compound of formula (I) according to any one of claims 1 to 4, wherein the 2θ angle error range is ±0.

20.

6. A method for preparing crystal form A of the compound of formula (I) according to any one of claims 1-5, comprising the following steps: The compound shown in formula (I) is mixed with solvent I, stirred to crystallize, and then separated into solid and liquid components. Solvent I is selected from one or more of water, ester solvents, ketone solvents, ether solvents, and hydrocarbon solvents. Preferably, the ester solvent is ethyl acetate or isopropyl acetate, the ketone solvent is selected from 2-butanone or methyl isobutyl ketone, the ether solvent is selected from tetrahydrofuran, and the hydrocarbon solvent is selected from n-heptane or cyclohexane.

7. A method for preparing crystal form B of the compound of formula (I) according to any one of claims 1-5, comprising the following steps: The compound shown in formula (I) is dissolved in an ether solvent, volatilized and crystallized, and then the solid and liquid are separated; preferably, the ether solvent is 1,4-dioxane.

8. A method for preparing crystal form C of the compound of formula (I) according to any one of claims 1-5, wherein the method is selected from the following: (i) Dissolve the compound shown in formula (I) in an ether solvent, volatilize and crystallize, and separate the solid and liquid phases; preferably, the ether solvent is propylene glycol methyl ether; (ii) The compound shown in formula (I) is mixed with a halohydrocarbon solvent, stirred to crystallize, and then separated into solid and liquid components; preferably, the halohydrocarbon solvent is chloroform; (iii) Dissolve the compound shown in formula (I) in solvent II to form solution 1, mix solution 1 with solvent III, stir to precipitate crystals, and separate the solid and liquid components. Solvent II is selected from ether solvents, and solvent III is selected from one or more of water, ether solvents, and hydrocarbon solvents. Preferably, the ether solvent in solvent II is selected from tetrahydrofuran, 1,4-dioxane or a mixture thereof; the ether solvent in solvent III is methyl tert-butyl ether, and the hydrocarbon solvent is n-heptane.

9. A method for preparing crystal form D of the compound of formula (I) according to any one of claims 1-5, wherein the method is selected from the following: (i) Dissolve the compound shown in formula (I) in solvent IV to form solution 1, mix solution 1 with solvent V, stir to precipitate crystals, and separate solid and liquid. Solvent IV is selected from ketone solvents, mixed solvents of ketone solvent and water, or halogenated hydrocarbon solvents. Solvent V is selected from water, hydrocarbon solvents, or ether solvents. Preferably, the ketone solvent in solvent IV is acetone, and the halohydrocarbon solvent is dichloromethane; the hydrocarbon solvent in solvent V is n-heptane, and the ether solvent is methyl tert-butyl ether. (ii) Mix the compound shown in formula (I) with solvent VI, stir to precipitate crystals, and separate the solid and liquid. Solvent VI is selected from ketone solvents or halogenated hydrocarbon solvents. Preferably, the ketone solvent in solvent VI is acetone, and the halogenated hydrocarbon solvent is dichloromethane.

10. A method for preparing crystal form E of the compound of formula (I) according to any one of claims 1-5, comprising the following steps: The compound shown in formula (I) is mixed with an ether solvent, stirred to induce crystallization, and then separated into solid and liquid components; preferably, the ether solvent is methyl tert-butyl ether.

11. A method for preparing crystal form F of the compound of formula (I) according to any one of claims 1-5, comprising the following steps: The compound shown in formula (I) is mixed with an ether solvent, stirred to crystallize, and then separated into solid and liquid components. Preferably, the ether solvent is propylene glycol methyl ether.

12. A method for preparing crystal form G of the compound of formula (I) according to any one of claims 1-5, wherein the crystal form is selected from the following methods: (i) The compound shown in formula (I) is mixed with solvent VII, stirred to induce crystallization, and then separated into solid and liquid components. Solvent VII is selected from water, alcohol solvents, ketone solvents, halogenated hydrocarbon solvents, ester solvents, and a mixture of hydrocarbon solvents. Preferably, the alcohol solvent is isopropanol, the ester solvent is ethyl acetate, the ketone solvent is acetone, the halogenated hydrocarbon solvent is selected from dichloromethane or trichloromethane, and the hydrocarbon solvent is n-heptane. (ii) The compound shown in formula (I) is mixed with solvent VIII, and the mixture is volatilized and crystallized, followed by solid-liquid separation. Solvent VIII is selected from a mixture of ketone solvents, halogenated hydrocarbon solvents, nitrile solvents, water, and alcohol solvents. Preferably, the ketone solvent is acetone, the halogenated hydrocarbon solvent is selected from dichloromethane or trichloromethane, the nitrile solvent is acetonitrile, and the alcohol solvent is isopropanol. (iii) Dissolve the compound shown in formula (I) in solvent IX to form solution 1, and mix solution 1 with solvent X, wherein solvent IX is selected from alcohol solvents, ether solvents, ketone solvents, halogenated hydrocarbon solvents or mixtures thereof, and solvent X is selected from hydrocarbon solvents, water, and ether solvents; preferably, the alcohol solvent in solvent IX is ethanol, the ether solvent is tetrahydrofuran, the ketone solvent is acetone, the halogenated hydrocarbon solvent is dichloromethane, the hydrocarbon solvent in solvent X is selected from n-heptane, and the ether solvent is selected from methyl tert-butyl ether.

13. A method for preparing crystal form H of the compound of formula (I) according to any one of claims 1-5, comprising the following steps: mixing the compound of formula (I) with an ester solvent, evaporating and crystallizing, and separating the solid and liquid phases; preferably, the ester solvent is ethyl acetate.

14. A pharmaceutical composition comprising the crystal form of the compound of formula (I) according to any one of claims 1 to 5 and at least one pharmaceutically acceptable excipient.

15. A method for preparing a pharmaceutical composition, comprising the step of mixing a crystal form of the compound of formula (I) according to any one of claims 1 to 5 or a mixture thereof with a pharmaceutically acceptable excipient.

16. Use of the crystal form of the compound of formula (I) according to any one of claims 1 to 5, or a mixture thereof, or the pharmaceutical composition according to claim 14, in the preparation of a medicament for treating or preventing diseases or conditions associated with abnormal activity of serine / threonine kinases.

17. Use of the crystal form of the compound of formula (I) according to any one of claims 1 to 5, or a mixture thereof, or the pharmaceutical composition according to claim 14, in the preparation of a medicament for treating or preventing diseases or conditions associated with abnormal activity of CDK2.

18. Use of the crystal form of the compound of formula (I) according to any one of claims 1 to 5, or a mixture thereof, or the pharmaceutical composition according to claim 14, in the preparation of a medicament for treating or preventing diseases or conditions associated with abnormal activity of CDK4.

19. Use of the crystal form of the compound of formula (I) according to any one of claims 1 to 5, or a mixture thereof, or the pharmaceutical composition according to claim 14, in the preparation of a medicament for treating or preventing a disease or condition, said disease or condition being selected from proliferative diseases, inflammatory diseases, autoinflammatory diseases, autoimmune diseases, or infectious diseases; preferably, said disease or condition is a proliferative disease; most preferably, said proliferative disease is selected from breast cancer (e.g., triple-negative breast cancer or ER-negative, PR-negative and Her2-positive breast cancer or CDK4 / 6 inhibitor-resistant breast cancer), colorectal cancer, lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, prostate cancer, Ewing's sarcoma, osteoma, neuroblastoma, cervical cancer, ovarian cancer, gastric cancer, and liver cancer.

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