Salts, crystalline forms of nitrogen-containing fused heterocyclic compounds, methods of making, pharmaceutical compositions, and uses thereof

Abstract

The present application relates to salts, crystalline forms of nitrogen-containing fused heterocyclic compounds, and methods of preparation, pharmaceutical compositions and uses thereof, in particular to salts, crystalline forms of compound I as shown below, and methods of preparation, pharmaceutical compositions and uses thereof. The salts of compound I or crystalline forms thereof exhibit at least one of the following advantages: improved bioavailability, good mechanical properties, improved chemical stability, excellent flow properties, good compressibility and improved dissolution characteristics.

Description

Technical Field

[0001] This invention relates to N-(4-(1-cyclopropyl-4-fluoro-2-methyl-1H-benzimidazol-6-yl)-5-fluoropyrimidin-2-yl)-6-(2-(dimethylamino)ethyl)-5,6,7,8-tetrahydro-1,6-naphthidine-2-amine), and more specifically, to the salts, crystal forms and preparation methods, pharmaceutical compositions and uses of the compound. Background Technology

[0002] Compound I, N-(4-(1-cyclopropyl-4-fluoro-2-methyl-1H-benzimidazol-6-yl)-5-fluoropyrimidin-2-yl)-6-(2-(dimethylamino)ethyl)-5,6,7,8-tetrahydro-1,6-naphthidine-2-amine, has the structure shown below: .

[0003] Compound I has been disclosed in CN106928219A and WO2017114512A1 (the entire contents of which are incorporated herein by reference, as if recorded herein). It exhibits high selectivity for cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6) at the molecular level, and high inhibitory activity against both CDK4 and CDK6. It also shows significant inhibitory effects on the proliferation of tumor cells associated with cyclin-dependent kinase activity at both the cellular and animal levels. It can be used to treat malignant tumors such as breast cancer, colon cancer, non-small cell lung cancer, astrocytoma, chronic myeloid leukemia, pancreatic cancer, acute monocytic leukemia, liver cancer (including hepatocellular carcinoma and hepatic adenocarcinoma), gastric cancer, non-small cell lung cancer, glioblastoma, and prostate adenocarcinoma. Furthermore, it exhibits good stability in human and mouse liver microsomes, no significant inhibition of metabolic enzymes, good absorption properties in mice and rats, high bioavailability, and good drug-like properties.

[0004] However, there is still a need for compound I that can provide desirable processing properties to improve the performance characteristics of the drug, such as ease of handling, ease of processing, improved solubility, improved shelf life, ease of purification, etc. Summary of the Invention

[0005] This invention provides a salt of compound I, its crystal form, a method for preparing the same, a pharmaceutical composition, and its uses. The salt of compound I or its crystal form exhibits advantages in at least one of the following aspects: improved bioavailability, good mechanical properties, improved chemical stability, excellent flow properties, good compressibility, and improved solubility characteristics.

[0006] On one hand, the present invention provides a salt of compound I as shown below, said salt being selected from maleate, hydrochloride, phosphate, lactate, fumarate, succinate, malate, adipate, hippurate, glycolate, benzoate, and nicotinate. .

[0007] The salt of compound I can be obtained by conventional methods for preparing salt compounds in the art, such as reacting the free base of compound I with a corresponding acid, or by acid displacement reaction, or by combining the methods described below and the methods disclosed in the examples with common knowledge in the art.

[0008] The salt may be one or more selected from the following crystal forms of compound I: maleate crystal form A, maleate crystal form B, maleate crystal form C, hydrochloride crystal form A, hydrochloride crystal form B, phosphate crystal form A, lactate crystal form A, fumarate crystal form A, succinate crystal form A, malate crystal form A, adipate crystal form A, hippurate crystal form A, glycolate crystal form A, benzoate crystal form A, and nicotinate crystal form A.

[0009] More specifically, the salt may be one or more selected from maleate crystal form A, fumarate crystal form A, adipate crystal form A and benzoate crystal form A of compound I.

[0010] More specifically, the salt can be maleate crystal form A of compound I.

[0011] More specifically, the salt can be the fumarate crystal form A of compound I.

[0012] More specifically, the salt can be the adipate crystal form A of compound I.

[0013] More specifically, the salt can be the benzoate crystal form A of compound I.

[0014] In one embodiment, the salt of compound I is a maleate.

[0015] In one embodiment, the maleate salt of compound I is present in the form of crystal form A, crystal form B, or crystal form C.

[0016] The X-ray powder diffraction (XRPD) pattern of the maleate crystal form A shows characteristic peaks at diffraction angles 2θ of approximately 5.48 ± 0.2°, 8.55 ± 0.2°, 10.92 ± 0.2°, 12.04 ± 0.2°, 12.98 ± 0.2°, 13.81 ± 0.2°, 16.40 ± 0.2°, 19.59 ± 0.2°, and 27.46 ± 0.2°. Preferably, characteristic peaks are also observed at 6.11 ± 0.2°, 7.84 ± 0.2°, 15.32 ± 0.2°, 18.70 ± 0.2°, 21.55 ± 0.2°, 22.21 ± 0.2°, 22.80 ± 0.2°, 23.32 ± 0.2°, 24.57 ± 0.2°, 25.62 ± 0.2°, and 26.07. Diffraction peaks are observed at one or more locations with diffraction angles of ±0.2°, 28.23 ±0.2°, 28.68 ±0.2°, 33.08 ±0.2°, and 38.76 ±0.2°, preferably at diffraction angles of approximately 5.48 ±0.2°, 6.11 ±0.2°, 7.84 ±0.2°, 8.55 ±0.2°, 10.92 ±0.2°, 12.04 ±0.2°, 12.98 ±0.2°, 13.81 ±0.2°, 15.32 ±0.2°, 16.40 ±0.2°, 18.70 ±0.2°, 19.59 ±0.2°, 21.55 ±0.2°, 22.21 ±0.2°, 22.80 ±0.2°, and 23.32. Characteristic peaks are observed at ±0.2°, 24.57±0.2°, 25.62 ±0.2°, 26.07 ±0.2°, 27.46 ±0.2°, 28.23 ±0.2°, 28.68±0.2°, 33.08 ±0.2°, and 38.76±0.2°. Specifically, the maleate crystal form A has essentially the following characteristics. Figure 1 The XRPD diagram shown is a Cu target used in the XRPD process.

[0017] The differential scanning calorimetry (DSC) plot of the maleate crystal form A has an endothermic peak with a starting temperature of approximately 222.8°C and an endothermic peak with a peak temperature of approximately 235.3°C. Specifically, the maleate crystal form A has substantially the following characteristics: Figure 2 The DSC diagram shown.

[0018] The thermogravimetric analysis (TGA) plot of maleate crystal form A shows that the sample loses approximately 4.1% of its weight at approximately 200°C, and the total weight loss in the range of approximately 200–290°C is approximately 17.5%, presumably due to deacidification (the theoretical weight loss for deacidification is approximately 18.7%). Specifically, maleate crystal form A has essentially the following characteristics: Figure 2 The TGA diagram shown is shown below.

[0019] through 1¹H NMR analysis showed that the molar ratio of maleic acid to free base in maleate crystal form A was approximately 1:1. Temperature-dependent XRPD (VT-XRPD) analysis revealed that maleate crystal form A remained unchanged after heating to approximately 170°C and cooling to approximately 30°C under N₂ protection, suggesting that maleate crystal form A is an amorphous form.

[0020] The X-ray powder diffraction (XRPD) pattern of the maleate crystal form B exhibits characteristic peaks at diffraction angles of approximately 5.75 ± 0.2°, 9.72 ± 0.2°, 14.91 ± 0.2°, 15.76 ± 0.2°, 17.43 ± 0.2°, 18.09 ± 0.2°, 22.20 ± 0.2°, 23.23 ± 0.2°, 25.17 ± 0.2°, and 27.96 ± 0.2°. In particular, the maleate crystal form B possesses essentially the following characteristics... Figure 3 The XRPD diagram shown is a Cu target used in the XRPD process.

[0021] The differential scanning calorimetry (DSC) plot of maleate crystal form B exhibits two weak endothermic peaks with peak temperatures of approximately 74.6 °C and 236.8 °C, respectively, and a sharp endothermic peak with an onset temperature of approximately 225.5 °C. Specifically, maleate crystal form B has substantially the following characteristics: Figure 4 The DSC diagram shown.

[0022] The thermogravimetric analysis (TGA) plot of maleate crystal form B shows a weight loss of 6.4% when the sample is heated to approximately 200 °C, and a total weight loss of approximately 19.1% in the range of approximately 200–290 °C, presumably due to deacidification (the theoretical weight loss for deacidification is approximately 18.7%). Specifically, maleate crystal form B has essentially the following characteristics: Figure 4 The TGA diagram shown is shown below.

[0023] through 1 According to H NMR analysis, the maleate crystal form B contains maleic acid in a molar ratio of approximately 1:1 to free base and contains a small amount of antisolvent, such as acetone or toluene.

[0024] The X-ray powder diffraction (XRPD) pattern of the maleate crystal form C exhibits characteristic peaks at diffraction angles of approximately 2θ: 5.51 ± 0.2°, 6.31 ± 0.2°, 8.91 ± 0.2°, 9.62 ± 0.2°, 10.59 ± 0.2°, 11.05 ± 0.2°, 12.17 ± 0.2°, 14.99 ± 0.2°, 15.73 ± 0.2°, 16.65 ± 0.2°, 21.40 ± 0.2°, 22.94 ± 0.2°, 24.71 ± 0.2°, 26.72 ± 0.2°, 28.22 ± 0.2°, 32.36 ± 0.2°, and 38.52 ± 0.2°. In particular, the maleate crystal form C has essentially the following characteristics... Figure 5 The XRPD diagram shown is a Cu target used in the XRPD process.

[0025] The differential scanning calorimetry (DSC) plot of the maleate crystal form C exhibits an exothermic peak with a peak temperature of approximately 142.8 °C and a sharp endothermic peak with a starting temperature of approximately 222.0 °C. In particular, the maleate crystal form C has substantially the following characteristics: Figure 6 The DSC diagram shown.

[0026] The thermogravimetric analysis (TGA) plot of the maleate crystal form C shows a weight loss of 6.3% when the sample is heated to approximately 200 °C, and a total weight loss of approximately 17.5% in the range of approximately 200–290 °C, presumably due to deacidification (the theoretical weight loss for deacidification is approximately 18.7%). Specifically, the maleate crystal form C has essentially the following characteristics: Figure 6 The TGA diagram shown is shown below.

[0027] through 1 ¹H NMR analysis showed that the molar ratio of maleic acid to free base in maleate crystal form C was approximately 1:1, with no solvent residue. VT-XRPD analysis revealed that maleate crystal form C remained unchanged when heated to approximately 110°C under N2 protection, suggesting that maleate crystal form C is an amorphous form.

[0028] Maleate crystal form B transforms into maleate crystal form C after being purged with N2 at approximately 30 °C, suggesting that maleate crystal form B is a hydrate. Maleate crystal forms B and C can also transform into maleate crystal form A.

[0029] The present invention also provides a method for preparing the above-mentioned maleate crystal form A, wherein the method is one of the following methods: Method 1: (1) The free base of compound I and maleic acid are reacted in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), alcohol solvents (e.g., methanol (MeOH)) and ester solvents (e.g., ethyl acetate (EtOA)); Method 2: (2) It is obtained by conversion of maleate crystal form B or C.

[0030] Both maleate crystal forms B and C are thermodynamically unstable crystals and will eventually transform into maleate crystal form A. Therefore, there are no particular limitations on the methods for converting maleate crystal forms B or C into maleate crystal form A. For example, maleate crystal forms B or C can be transformed into maleate crystal form A by suspending and stirring in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), and ester solvents (e.g., ethyl acetate (EtOA)), but the present invention is not limited thereto.

[0031] The present invention also provides a method for preparing the above-mentioned maleate crystal form B, the method comprising: recrystallizing the maleate of compound I by adding an antisolvent (e.g., acetone and toluene) dropwise to a mixed solution of methanol (MeOH) / dichloromethane (DCM) (1:1, v / v), and then placing the resulting solid under room temperature and humidity.

[0032] The present invention also provides a method for preparing the above-mentioned maleate crystal form C, the method comprising: purging maleate crystal form B with an inert gas (e.g., N2) to obtain it.

[0033] In one embodiment, the salt of compound I is a hydrochloride salt.

[0034] In one embodiment, the hydrochloride salt of compound I exists in crystal form A or crystal form B.

[0035] The X-ray powder diffraction (XRPD) pattern of the hydrochloride crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.41 ± 0.2°, 5.34 ± 0.2°, 8.90 ± 0.2°, 9.16 ± 0.2°, 10.69 ± 0.2°, 11.07 ± 0.2°, 13.16 ± 0.2°, 15.63 ± 0.2°, 18.49 ± 0.2°, 19.25 ± 0.2°, 21.28 ± 0.2°, 24.60 ± 0.2°, and 26.85 ± 0.2°. Specifically, the XRPD pattern of the hydrochloride crystal form A is essentially as follows: Figure 7 As shown, the target used in the XRPD is a Cu target.

[0036] The differential scanning calorimetry (DSC) plot of the hydrochloride crystal form A has endothermic peaks with peak temperatures of approximately 94.6°C, approximately 237.4°C, and approximately 266.9°C, respectively. In particular, the hydrochloride crystal form A has substantially the following characteristics: Figure 8 The DSC diagram shown.

[0037] The thermogravimetric analysis (TGA) plot of the hydrochloride crystal form A shows that the sample experiences a weight loss of approximately 3.5% when heated to approximately 100°C. Specifically, the hydrochloride crystal form A has essentially the following characteristics: Figure 8 The TGA diagram shown is shown below.

[0038] through 1 According to H NMR analysis, in the hydrochloride crystal form A, the molar ratio of hydrochloric acid to free base is approximately 1:1.

[0039] The present invention also provides a method for preparing the above-mentioned hydrochloride crystal form A, the method comprising: reacting a free base of compound I and hydrochloric acid in a molar ratio of about 1:1 in a solvent selected from ether solvents (e.g., tetrahydrofuran (THF)) and ester solvents (e.g., ethyl acetate (EtOA)).

[0040] The X-ray powder diffraction (XRPD) pattern of the hydrochloride crystal form B exhibits characteristic peaks at diffraction angles of approximately 3.73 ± 0.2°, 4.96 ± 0.2°, 7.24 ± 0.2°, 10.26 ± 0.2°, and 15.47 ± 0.2°. Specifically, the XRPD pattern of the hydrochloride crystal form B is essentially as follows: Figure 7 As shown, the target type used in the XRPD is a Cu target.

[0041] The differential scanning calorimetry (DSC) plot of the hydrochloride crystal form B has endothermic peaks with peak temperatures of approximately 109.9°C, approximately 160.3°C, and approximately 266.9°C, respectively. In particular, the hydrochloride crystal form B has substantially the following characteristics: Figure 9 The DSC diagram shown is shown below.

[0042] The thermogravimetric analysis (TGA) plot of the hydrochloride crystal form B shows that the sample experiences a weight loss of approximately 9.1% upon heating to approximately 130°C. Specifically, the hydrochloride crystal form B has essentially the following characteristics: Figure 9 The TGA diagram shown is shown below.

[0043] through 1 According to H NMR analysis, the molar ratio of hydrochloric acid to free base in the hydrochloride crystal form B is approximately 3:1.

[0044] The present invention also provides a method for preparing the above-mentioned hydrochloride crystal form B, the method comprising: reacting a free base of compound I and hydrochloric acid in a molar ratio of about 3:1 in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), ester solvents (e.g., ethyl acetate (EtOA)), and mixed solvents (e.g., haloalkanes and alcohol solvents, such as dichloromethane / MeOH).

[0045] In one embodiment, the salt of compound I is a phosphate.

[0046] In one embodiment, the phosphate of compound I exists in crystal form A.

[0047] The X-ray powder diffraction (XRPD) pattern of the phosphate crystal form A exhibits characteristic peaks at diffraction angles of approximately 5.63 ± 0.2°, 7.86 ± 0.2°, 9.88 ± 0.2°, 12.43 ± 0.2°, 15.86 ± 0.2°, 19.64 ± 0.2°, 21.26 ± 0.2°, 22.72 ± 0.2°, 25.42 ± 0.2°, and 27.32 ± 0.2°. In particular, the phosphate crystal form A has essentially the following characteristics... Figure 10 The XRPD diagram shown is a Cu target used in the XRPD process.

[0048] The differential scanning calorimetry (DSC) plot of the phosphate crystal form A has endothermic peaks with onset temperatures of approximately 48.0°C and approximately 228.3°C, respectively. In particular, the phosphate crystal form A has substantially the following characteristics: Figure 11 The DSC diagram shown is shown below.

[0049] The thermogravimetric analysis (TGA) plot of the phosphate crystal form A shows that the sample experiences a weight loss of approximately 3.6% upon heating to approximately 130°C. Specifically, the phosphate crystal form A has essentially the following characteristics: Figure 11 The TGA diagram shown is shown below.

[0050] through 1 According to H NMR analysis, in the phosphate crystal form A, the molar ratio of phosphoric acid to free base is approximately 1:1.

[0051] The present invention also provides a method for preparing the above-mentioned phosphate crystal form A, the method comprising: reacting a free base of compound I and phosphoric acid in a molar ratio of about 1:1 in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), ester solvents (e.g., ethyl acetate (EtOA)), and mixed solvents (e.g., haloalkanes and alcohol solvents, such as dichloromethane / MeOH).

[0052] In one embodiment, the salt of compound I is a lactate.

[0053] In one embodiment, the lactate of compound I exists in crystal form A.

[0054] The X-ray powder diffraction (XRPD) pattern of the lactate crystal form A shows characteristic peaks at diffraction angles of approximately 4.52 ± 0.2°, 5.12 ± 0.2°, 6.94 ± 0.2°, 8.97 ± 0.2°, 10.16 ± 0.2°, 10.49 ± 0.2°, 11.30 ± 0.2°, 13.35 ± 0.2°, 13.86 ± 0.2°, 17.51 ​​± 0.2°, 18.52 ± 0.2°, 21.02 ± 0.2°, 21.97 ± 0.2°, 25.10 ± 0.2°, 26.05 ± 0.2°, and 27.04 ± 0.2°. In particular, the lactate crystal form A has essentially the following characteristics... Figure 12 The XRPD diagram shown is a Cu target used in the XRPD process.

[0055] The differential scanning calorimetry (DSC) plot of the lactate crystal form A has endothermic peaks with peak temperatures of approximately 99.0 ºC and approximately 183.3 ºC, respectively. In particular, the lactate crystal form A has substantially the following characteristics: Figure 13 The DSC diagram shown is shown below.

[0056] The thermogravimetric analysis (TGA) plot of the lactate crystal form A shows that the sample experiences a weight loss of approximately 4.0% when heated to approximately 130°C. Specifically, the lactate crystal form A has essentially the following characteristics: Figure 13 The TGA diagram shown is shown below.

[0057] through 1 According to H NMR analysis, the molar ratio of lactic acid to free base in the lactate crystal form A is approximately 1:1.

[0058] The present invention also provides a method for preparing the above-mentioned lactate crystal form A, the method comprising: reacting compound I free base and lactic acid in a molar ratio of about 1:1 in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), and ester solvents (e.g., ethyl acetate (EtOA)).

[0059] In one embodiment, the salt of compound I is a fumarate.

[0060] In one embodiment, the fumarate of compound I exists in crystal form A.

[0061] The X-ray powder diffraction (XRPD) pattern of the fumarate crystal form A exhibits characteristic peaks at diffraction angles of approximately 5.73 ± 0.2°, 6.29 ± 0.2°, 8.04 ± 0.2°, 10.28 ± 0.2°, 11.27 ± 0.2°, 12.79 ± 0.2°, 14.17 ± 0.2°, 14.99 ± 0.2°, 16.07 ± 0.2°, 17.28 ± 0.2°, 18.16 ± 0.2°, 19.90 ± 0.2°, 20.62 ± 0.2°, 22.13 ± 0.2°, 23.10 ± 0.2°, 23.82 ± 0.2°, 24.52 ± 0.2°, and 27.03 ± 0.2°. In particular, the fumarate crystal form A has essentially the following characteristics... Figure 14 The XRPD diagram shown is a Cu target used in the XRPD process.

[0062] The differential scanning calorimetry (DSC) plot of the fumarate crystal form A shows an exothermic peak with a peak temperature of approximately 163.5 °C and an endothermic peak with an onset temperature of approximately 248.9 °C. Specifically, the fumarate crystal form A has substantially the following characteristics: Figure 15 The DSC diagram shown.

[0063] The thermogravimetric analysis (TGA) plot of fumarate crystal form A shows that the sample experiences a weight loss of approximately 2.26% upon heating to approximately 130°C. Specifically, fumarate crystal form A has essentially the following characteristics: Figure 15 The TGA diagram shown is shown below.

[0064] through 1 According to H NMR analysis, the molar ratio of fumaric acid to free base in the fumarate crystal form A is approximately 1:1.

[0065] The present invention also provides a method for preparing the above-mentioned fumarate crystal form A, the method comprising: reacting a free base of compound I and fumaric acid in a molar ratio of about 1:1 in a solvent selected from ether solvents (e.g., tetrahydrofuran (THF)), ester solvents (e.g., ethyl acetate (EtOA)), and mixed solvents (e.g., haloalkanes and alcohol solvents, such as dichloromethane / MeOH).

[0066] In one embodiment, the salt of compound I is a succinate.

[0067] In one embodiment, the succinate of compound I exists in crystal form A.

[0068] The X-ray powder diffraction (XRPD) pattern of the succinate crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.18 ± 0.2°, 5.33 ± 0.2°, 6.82 ± 0.2°, 8.35 ± 0.2°, 11.56 ± 0.2°, 13.67 ± 0.2°, 16.44 ± 0.2°, 17.74 ± 0.2°, 20.47 ± 0.2°, and 23.15 ± 0.2°. In particular, the succinate crystal form A has essentially the following characteristics... Figure 16 The XRPD diagram shown is a Cu target used in the XRPD process.

[0069] The differential scanning calorimetry (DSC) plot of the succinate crystal form A has endothermic peaks with onset temperatures of approximately 51.6°C and approximately 182.1°C, respectively. In particular, the succinate crystal form A has substantially the following characteristics: Figure 17 The DSC diagram shown.

[0070] The thermogravimetric analysis (TGA) plot of succinate crystal form A shows that the sample experiences a weight loss of approximately 3.2% upon heating to approximately 130°C. Specifically, succinate crystal form A has essentially the following characteristics: Figure 17 The TGA diagram shown is shown below.

[0071] through 1 According to H NMR analysis, the molar ratio of succinic acid to free base in the succinate crystal form A is approximately 1:1.

[0072] The present invention also provides a method for preparing the above-mentioned succinate crystal form A, the method comprising: reacting a free base of compound I and succinic acid in a molar ratio of about 1:1 in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), and ester solvents (e.g., ethyl acetate (EtOA)).

[0073] In one embodiment, the salt of compound I is malate.

[0074] In one embodiment, the malate of compound I exists in crystal form A.

[0075] The X-ray powder diffraction (XRPD) pattern of the malate crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.49 ± 0.2°, 6.10 ± 0.2°, 7.16 ± 0.2°, 9.00 ± 0.2°, 10.99 ± 0.2°, 14.87 ± 0.2°, 16.65 ± 0.2°, 19.73 ± 0.2°, 20.55 ± 0.2°, 21.96 ± 0.2°, 23.12 ± 0.2°, 24.03 ± 0.2°, 25.87 ± 0.2°, and 26.58 ± 0.2°. In particular, the malate crystal form A has essentially the following characteristics... Figure 18 The XRPD diagram shown is a Cu target used in the XRPD process.

[0076] The differential scanning calorimetry (DSC) plot of the malate crystal form A has endothermic peaks with peak temperatures of approximately 100.4°C, approximately 175.9°C, and approximately 185.6°C, respectively. In particular, the malate crystal form A has substantially the following characteristics: Figure 19 The DSC diagram shown.

[0077] The thermogravimetric analysis (TGA) plot of the malate crystal form A shows that the sample experiences a weight loss of approximately 4.0% upon heating to approximately 130°C. Specifically, the malate crystal form A has essentially the following characteristics: Figure 19 The TGA diagram shown is shown below.

[0078] through 1 According to H NMR analysis, the molar ratio of malic acid to free base in the malate crystal form A is approximately 1:1.

[0079] The present invention also provides a method for preparing the above-mentioned malate crystal form A, the method comprising: reacting a free base of compound I and malic acid in a molar ratio of about 1:1 in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), and ester solvents (e.g., ethyl acetate (EtOA)).

[0080] In one embodiment, the salt of compound I is adipic acid salt.

[0081] In one embodiment, the adipate of compound I exists in crystal form A.

[0082] The X-ray powder diffraction (XRPD) pattern of the adipate crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.53 ± 0.2°, 4.85 ± 0.2°, 6.03 ± 0.2°, 7.15 ± 0.2°, 9.05 ± 0.2°, 9.56 ± 0.2°, 10.94 ± 0.2°, 12.18 ± 0.2°, 13.66 ± 0.2°, 14.61 ± 0.2°, 18.23 ± 0.2°, 20.12 ± 0.2°, 24.05 ± 0.2°, 25.77 ± 0.2°, 26.25 ± 0.2°, and 27.50 ± 0.2°. In particular, the adipate crystal form A has essentially the following characteristics... Figure 20 The XRPD diagram shown is a Cu target used in the XRPD process.

[0083] The differential scanning calorimetry (DSC) plot of the adipate crystal form A shows an endothermic peak with a starting temperature of approximately 182.0ºC. Specifically, the adipate crystal form A has substantially the following characteristics: Figure 21The DSC diagram shown.

[0084] The thermogravimetric analysis (TGA) plot of adipate crystal form A shows that the sample experiences a weight loss of approximately 2.5% upon heating to approximately 130°C. Specifically, adipate crystal form A has essentially the following characteristics: Figure 21 The TGA diagram shown is shown below.

[0085] through 1 According to H NMR analysis, in the adipic acid salt crystal form A, the molar ratio of adipic acid to free base is approximately 1:1.

[0086] The present invention also provides a method for preparing the above-mentioned adipate crystal form A, the method comprising: reacting a free base of compound I and adipic acid in a molar ratio of about 1:1 in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), and ester solvents (e.g., ethyl acetate (EtOA)).

[0087] In one embodiment, the salt of compound I is hippurate.

[0088] In one embodiment, the hippurate of compound I exists in crystal form A.

[0089] The X-ray powder diffraction (XRPD) pattern of hippurate crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.45 ± 0.2°, 5.50 ± 0.2°, 9.17 ± 0.2°, 18.79 ± 0.2°, 23.20 ± 0.2°, and 25.42 ± 0.2°. In particular, hippurate crystal form A possesses essentially the following characteristics... Figure 22 The XRPD diagram shown is a Cu target used in the XRPD process.

[0090] The differential scanning calorimetry (DSC) plot of the hippurate crystal form A has endothermic peaks with peak temperatures of approximately 124.2°C, approximately 141.0°C, approximately 154.6°C, approximately 178.0°C, and approximately 217.2°C, respectively. In particular, the hippurate crystal form A has substantially the following characteristics: Figure 23 The DSC diagram shown.

[0091] The thermogravimetric analysis (TGA) plot of hippurate crystal form A shows that the sample experiences a weight loss of approximately 4.2% upon heating to approximately 130°C. Specifically, hippurate crystal form A has essentially the following properties: Figure 23 The TGA diagram shown is shown below.

[0092] through 1 According to H NMR analysis, the molar ratio of hippuric acid to free base in hippurate crystal form A is approximately 1:1.

[0093] The present invention also provides a method for preparing the above-mentioned hippurate crystal form A, the method comprising: reacting compound I free base and hippuric acid in a molar ratio of about 1:1 in a solvent selected from ketone solvents (e.g., acetone) and ester solvents (e.g., ethyl acetate (EtOA)).

[0094] In one embodiment, the salt of compound I is a glycolate.

[0095] In one embodiment, the glycolate of compound I exists in crystal form A.

[0096] The X-ray powder diffraction (XRPD) pattern of the glycolate crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.50 ± 0.2°, 5.19 ± 0.2°, 6.90 ± 0.2°, 9.00 ± 0.2°, 10.42 ± 0.2°, 11.35 ± 0.2°, 13.74 ± 0.2°, 15.60 ± 0.2°, 16.24 ± 0.2°, 17.97 ± 0.2°, 20.77 ± 0.2°, 22.68 ± 0.2°, 25.12 ± 0.2°, 26.61 ± 0.2°, 27.69 ± 0.2°, and 31.73 ± 0.2°. In particular, the glycolate crystal form A has essentially the following characteristics... Figure 24 The XRPD diagram shown is a Cu target used in the XRPD process.

[0097] The differential scanning calorimetry (DSC) plot of the glycolate crystal form A has endothermic peaks with onset temperatures of approximately 54.1°C and approximately 183.2°C, respectively. In particular, the glycolate crystal form A has substantially the following characteristics: Figure 25 The DSC diagram shown is shown below.

[0098] The thermogravimetric analysis (TGA) plot of glycolate crystal form A shows that the sample experiences a weight loss of approximately 4.8% upon heating to approximately 130°C. Specifically, glycolate crystal form A has essentially the following characteristics: Figure 25 The TGA diagram shown is shown below.

[0099] through 1 According to H NMR analysis, in the glycolate crystal form A, the molar ratio of glycolic acid to free base is approximately 1:1.

[0100] The present invention also provides a method for preparing the above-mentioned glycolate crystal form A, the method comprising: reacting a free base of compound I and glycolic acid in a molar ratio of about 1:1 in a solvent selected from ketone solvents (e.g., acetone), ether solvents (e.g., tetrahydrofuran (THF)), and ester solvents (e.g., ethyl acetate (EtOA)).

[0101] In one embodiment, the salt of compound I is a benzoate.

[0102] In one embodiment, the benzoate of compound I exists in crystal form A.

[0103] The X-ray powder diffraction (XRPD) pattern of the benzoate crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.47 ± 0.2°, 4.80 ± 0.2°, 6.74 ± 0.2°, 8.96 ± 0.2°, 9.94 ± 0.2°, 10.40 ± 0.2°, 12.78 ± 0.2°, 13.50 ± 0.2°, 14.88 ± 0.2°, 17.41 ± 0.2°, 18.32 ± 0.2°, 19.54 ± 0.2°, 20.84 ± 0.2°, 23.58 ± 0.2°, 25.01 ± 0.2°, 26.31 ± 0.2°, 27.11 ± 0.2°, and 29.33 ± 0.2°. In particular, the benzoate crystal form A has essentially the following characteristics... Figure 26 The XRPD diagram shown is a Cu target used in the XRPD process.

[0104] The differential scanning calorimetry (DSC) plot of the benzoate crystal form A shows an endothermic peak with a starting temperature of approximately 169.5ºC. Specifically, the benzoate crystal form A has substantially the following characteristics: Figure 27 The DSC diagram shown is shown below.

[0105] The thermogravimetric analysis (TGA) plot of benzoate crystal form A shows that the sample experiences a weight loss of approximately 2.7% upon heating to approximately 130°C. Specifically, benzoate crystal form A has essentially the following characteristics: Figure 27 The TGA diagram shown is shown below.

[0106] through 1 According to H NMR analysis, in the benzoate crystal form A, the molar ratio of benzoic acid to free base is approximately 1:1.

[0107] The present invention also provides a method for preparing the above-mentioned benzoate crystal form A, the method comprising: reacting compound I free base and benzoic acid in a molar ratio of about 1:1 in a ketone solvent (e.g., acetone) to obtain the product.

[0108] In one embodiment, the salt of compound I is nicotinic acid salt.

[0109] In one embodiment, the nicotinate of compound I exists in crystal form A.

[0110] The X-ray powder diffraction (XRPD) pattern of the nicotinate crystal form A exhibits characteristic peaks at diffraction angles of approximately 4.54±0.2°, 6.46±0.2°, 10.14±0.2°, 13.41±0.2°, 14.66±0.2°, 17.14±0.2°, 18.83±0.2°, 21.36±0.2°, 24.83±0.2°, and 26.27±0.2°. In particular, the nicotinate crystal form A has essentially the following characteristics... Figure 28 The XRPD diagram shown is a Cu target used in the XRPD process.

[0111] The differential scanning calorimetry (DSC) plot of the nicotinate crystal form A has endothermic peaks with peak temperatures of approximately 103.9°C, approximately 160.3°C, and approximately 212.5°C, respectively. In particular, the nicotinate crystal form A has substantially the following characteristics: Figure 29 The DSC diagram shown is shown below.

[0112] The thermogravimetric analysis (TGA) plot of the nicotinate crystal form A shows that the sample experiences a weight loss of approximately 4.9% upon heating to approximately 130°C. Specifically, the nicotinate crystal form A has essentially the following characteristics: Figure 29 The TGA diagram shown is shown below.

[0113] through 1 According to H NMR analysis, in the nicotinic acid salt crystal form A, the molar ratio of nicotinic acid to free base is approximately 1:1.

[0114] The present invention also provides a method for preparing the above-mentioned nicotinic acid salt crystal form A, the method comprising: reacting compound I free base and nicotinic acid in a molar ratio of about 1:1 in a solvent selected from ether solvents (e.g., tetrahydrofuran (THF)) and ester solvents (e.g., ethyl acetate (EtOA)).

[0115] In the method for preparing the crystal form of the present invention, seed crystals may also be used. Seed crystals may be used in the methods for preparing the various crystal forms of the present application according to methods known in the art.

[0116] In another aspect, the present invention provides a pharmaceutical composition comprising one or more salts selected from the above-described compound I, said salt being selected from maleate, hydrochloride, phosphate, lactate, fumarate, succinate, malate, adipate, hippurate, glycolate, benzoate, and nicotinate.

[0117] In addition, the pharmaceutical composition may also include a pharmaceutically acceptable carrier. The choice of a pharmaceutically acceptable carrier varies depending on the route of administration and characteristics of action, and is typically a filler, diluent, binder, wetting agent, disintegrant, lubricant, emulsifier, suspending agent, etc., which are conventional in the art.

[0118] The pharmaceutical composition can be administered orally, by injection (intravenous, intramuscular, subcutaneous, and intracoronary), sublingually, buccally, rectally, urethra, vaginally, nasally, by inhalation, or topically, with oral administration being the preferred route.

[0119] The pharmaceutical composition can be used to prevent or treat diseases associated with abnormal cell cycle regulation. The term "diseases associated with abnormal cell cycle regulation" can refer to "diseases associated with abnormalities in cyclin-dependent kinases (preferably CDK4 and / or CDK6)," particularly tumors, and more particularly malignant tumors (e.g., breast cancer, colon cancer, non-small cell carcinoma, astrocytoma, chronic myeloid leukemia, pancreatic cancer, acute monocytic leukemia, liver cancer (including hepatocellular carcinoma, hepatic adenocarcinoma), gastric cancer, non-small cell lung cancer, glioblastoma, and prostate adenocarcinoma), and advanced solid tumors (including but not limited to breast cancer, primary / metastatic tumors of the central nervous system, etc.).

[0120] The pharmaceutical composition can be used as a cyclin-dependent kinase (preferably CDK4 and / or CDK6) inhibitor.

[0121] The pharmaceutical composition can be used to inhibit the proliferation of tumor cells. The tumor cells are preferably cancer cells; the cancer cells are preferably breast cancer cells, colon cancer cells, non-small cell carcinoma cells, brain astrocytoma cells, chronic myeloid leukemia cells, pancreatic cancer cells, acute monocytic leukemia cells, liver cancer cells (including hepatocellular carcinoma cells and hepatic adenocarcinoma cells), gastric cancer cells, non-small cell lung cancer cells, glioblastoma cells, and prostate adenocarcinoma cells; the breast cancer cells are preferably one or more of breast cancer cells MCF-7, T-47D, and ZR-75-1.

[0122] On the other hand, the present invention provides the use of a salt of the above-mentioned compound I for the preparation of a medicament, said salt being selected from maleate, hydrochloride, phosphate, lactate, fumarate, succinate, malate, adipic acidate, hippurate, glycolate, benzoate, and nicotinate, said medicament for the prevention or treatment of diseases associated with abnormal cell cycle regulation. Specifically, the "diseases associated with abnormal cell cycle regulation" can be "diseases associated with abnormalities in cyclin-dependent kinases (preferably CDK4 and / or CDK6)," particularly tumors, more particularly malignant tumors (e.g., breast cancer, colon cancer, non-small cell carcinoma, astrocytoma, chronic myeloid leukemia, pancreatic cancer, acute monocytic leukemia, liver cancer (including hepatocellular carcinoma, hepatic adenocarcinoma), gastric cancer, non-small cell lung cancer, glioblastoma, and prostate adenocarcinoma), and advanced solid tumors (including but not limited to breast cancer, primary / metastatic tumors of the central nervous system, etc.).

[0123] In another aspect, the present invention provides the use of salts of the above-mentioned compound I for the preparation of inhibitors of cyclin-dependent kinases (preferably CDK4 and / or CDK6), wherein the salts are selected from maleate, hydrochloride, phosphate, lactate, fumarate, succinate, malate, adipate, hippurate, glycolate, benzoate and nicotinate.

[0124] In another aspect, the present invention provides the use of a salt of the above-described compound I for preparing a medicament that inhibits the proliferation of tumor cells, said salt being selected from maleate, hydrochloride, phosphate, lactate, fumarate, succinate, malate, adipate, hippurate, glycolate, benzoate, and nicotinate. The tumor cells are preferably cancer cells; the cancer cells are preferably breast cancer cells; the breast cancer cells are preferably one or more selected from breast cancer cells MCF-7, T-47D, and ZR-75-1.

[0125] In the above-described pharmaceutical compositions and uses, the salt may be one or more selected from the following crystal forms of compound I: maleate crystal form A, maleate crystal form B, maleate crystal form C, hydrochloride crystal form A, hydrochloride crystal form B, phosphate crystal form A, lactate crystal form A, fumarate crystal form A, succinate crystal form A, malate crystal form A, adipate crystal form A, hippurate crystal form A, glycolate crystal form A, benzoate crystal form A, and nicotinate crystal form A.

[0126] More specifically, the salt may be one or more selected from maleate crystal form A, fumarate crystal form A, adipate crystal form A and benzoate crystal form A of compound I.

[0127] More specifically, the salt can be maleate crystal form A of compound I.

[0128] More specifically, the salt can be the fumarate crystal form A of compound I.

[0129] More specifically, the salt can be the adipate crystal form A of compound I.

[0130] More specifically, the salt can be the benzoate crystal form A of compound I.

[0131] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0132] Figure 1 This is an XRPD diagram of maleate crystal form A of compound I according to the present invention.

[0133] Figure 2These are the DSC and TGA diagrams of maleate crystal form A of compound I according to the present invention.

[0134] Figure 3 This is an XRPD diagram of maleate crystal form B of compound I according to the present invention.

[0135] Figure 4 These are the DSC and TGA diagrams of maleate crystal form B of compound I according to the present invention.

[0136] Figure 5 This is an XRPD diagram of maleate crystal form C of compound I according to the present invention.

[0137] Figure 6 These are the DSC and TGA diagrams of maleate crystal form C of compound I according to the present invention.

[0138] Figure 7 These are XRPD diagrams of crystal forms A and B of compound I hydrochloride according to the present invention.

[0139] Figure 8 These are DSC and TGA diagrams of crystal form A of compound I hydrochloride according to the present invention.

[0140] Figure 9 These are the DSC and TGA diagrams of crystal form B of compound I hydrochloride according to the present invention.

[0141] Figure 10 This is an XRPD diagram of phosphate crystal form A of compound I according to the present invention.

[0142] Figure 11 These are DSC and TGA diagrams of phosphate crystal form A of compound I according to the present invention.

[0143] Figure 12 This is an XRPD diagram of compound I lactate crystal form A according to the present invention.

[0144] Figure 13 These are the DSC and TGA diagrams of compound I lactate crystal form A according to the present invention.

[0145] Figure 14 This is an XRPD diagram of the fumarate crystal form A of compound I according to the present invention.

[0146] Figure 15 These are the DSC and TGA diagrams of compound I fumarate crystal form A according to the present invention.

[0147] Figure 16 This is an XRPD diagram of succinate crystal form A of compound I according to the present invention.

[0148] Figure 17These are the DSC and TGA diagrams of succinate crystal form A of compound I according to the present invention.

[0149] Figure 18 This is an XRPD diagram of compound I malate crystal form A according to the present invention.

[0150] Figure 19 These are the DSC and TGA diagrams of compound I malate crystal form A according to the present invention.

[0151] Figure 20 This is an XRPD diagram of compound I adipate crystal form A according to the present invention.

[0152] Figure 21 These are the DSC and TGA diagrams of compound I adipate crystal form A according to the present invention.

[0153] Figure 22 This is an XRPD diagram of the crystal form A of compound I hippurate according to the present invention.

[0154] Figure 23 These are the DSC and TGA diagrams of compound I hippurate crystal form A according to the present invention.

[0155] Figure 24 This is an XRPD diagram of the crystal form A of compound I glycolate according to the present invention.

[0156] Figure 25 The figures shown are DSC and TGA diagrams of crystal form A of compound I glycolate according to the present invention.

[0157] Figure 26 This is an XRPD diagram of benzoate form A of compound I according to the present invention.

[0158] Figure 27 These are the DSC and TGA diagrams of benzoate form A of compound I according to the present invention.

[0159] Figure 28 This is an XRPD diagram of nicotinate crystal form A of compound I according to the present invention.

[0160] Figure 29 These are the DSC and TGA diagrams of compound I nicotinate crystal form A according to the present invention. Detailed Implementation

[0161] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0162] Unless otherwise stated, the equipment and testing methods involved in the embodiments of this invention are as follows: X-ray powder diffraction (XRPD): XRPD patterns were acquired using a PANalytacal Empyrean X-ray powder diffractometer. XRPD parameters are shown in Table 1.

[0163] Table 1

[0164] Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC): TGA and DSC spectra were acquired on a TA Q500 / 5000 thermogravimetric analyzer and a TAQ200 / 2000 differential scanning calorimeter, respectively. The test parameters are shown in Table 2.

[0165] Table 2: DSC and TGA Test Parameters

[0166] High performance liquid chromatography (HPLC): High performance liquid chromatography was performed on an Agilent 1100 HPLC. Specific HPLC purity test parameters are shown in Table 3 below.

[0167] Table 3

[0168] Ion chromatography (IC): The content of negative ions was measured on an ion chromatograph (IC) Thermo ICS1100 and the molar ratio was determined by coupling with HPLC data. Specific instrument parameters are listed in Table 4.

[0169] Table 4

[0170] Dynamic moisture adsorption (DVS): Dynamic moisture adsorption (DVS) curves were acquired on the DVS Intrinsic of SMS (Surface Measurement Systems). Relative humidity at 25 ºC was corrected for the deliquescence points of LiCl, Mg(NO3)2, and KCl. DVS test parameters are listed in Table 5.

[0171] Table 5

[0172] Liquid NMR: Liquid NMR spectra were acquired on a Bruker 400M NMR spectrometer using DMSO-d6 as the solvent.

[0173] The heating test was conducted as follows: The sample was heated to the target temperature under nitrogen protection and then cooled to room temperature. The sample was then removed and exposed to air, and XRPD was collected for testing.

[0174] The free base of compound 1 was prepared according to the method disclosed in CN106928219A. Only a few diffraction peaks were observed in its XRPD results. The HPLC purity was 95.2% by area. TGA results showed a weight loss of 4.8% when the sample was heated to 200ºC. DSC results showed one endothermic and two exothermic signals before melting at 233.6ºC (initial temperature). Heating experiments showed no change in XRPD results when the initial sample was heated to 80ºC; the diffraction peaks at approximately 2θ of 4.7º disappeared when heated to 135ºC and 175ºC, indicating increased crystallinity.

[0175] Preparation Example 1 The crystal form was prepared using the following method: Acetone, tetrahydrofuran (THF), ethyl acetate (EtOA), and dichloromethane (DCM) / methanol (MeOH) (10:1, v / v) were used as solvents, respectively.

[0176] Weigh approximately 20 mg of the free base of compound I, add the corresponding acid according to the molar ratio in Table 6, add 0.3–0.5 mL of solvent, stir at room temperature for 2 days, centrifuge to separate the solid, and dry at 50 °C for 2 hours for purity characterization by XRPD, TGA, DSC, and HPLC. Blank indicates no acid was added.

[0177] The results of crystal form preparation and screening are shown in Table 6.

[0178] Table 6

[0179] XRPD results for free alkali crystal form A showed a few diffraction peaks, with an HPLC purity of 98.2% (area %). TGA results indicated a 1.2% weight loss when the sample was heated to 130ºC. DSC results showed an endothermic peak before melting at 239.9ºC (initial temperature). DVS results showed a 2.9% water adsorption capacity at 25ºC / 80%RH, and XRPD results before and after DVS testing showed no change in crystal form.

[0180] Repeated crystal form preparation revealed that the crystallinity of samples in the tartaric acid, citric acid, gentian acid, and methanesulfonic acid systems was relatively weak.

[0181] The safety level and crystallinity data of the acids used for each crystal form sample are summarized in Table 7 below.

[0182] Table 7

[0183] Note: +: Data not collected due to small sample size; --: Data not collected due to sample crystallinity and acid safety level.

[0184] The XRPD characterization results of maleate crystal form A are as follows: Figure 1 The XRPD diffraction peak data are shown in Table 8. The TGA results are as follows: Figure 2 The TGA curves show that maleate crystal form A loses 4.05% of its weight when heated to approximately 200 ºC. The DSC results are as follows: Figure 2 The DSC curves show melting at approximately 222.8 ºC (initial temperature) accompanied by decomposition. HPLC / IC results indicate a molar ratio of 1.03 (acid / base), suggesting that maleate crystal form A is an amorphous form of maleate.

[0185] Table 8: XRPD diffraction peak data of maleate crystal form A

[0186] The XRPD results of the fumarate crystal form A are as follows: Figure 14 As shown. TGA results are as follows. Figure 15 The TGA curves show that the fumarate crystal form A sample loses 2.3% of its weight when heated to approximately 130ºC. The DSC results are as follows... Figure 15 The DSC curves show an exothermic peak at approximately 163.5 ºC (peak temperature) and an endothermic peak at approximately 248.9 ºC (starting temperature). Heating experiments show that the crystal form remains unchanged when the sample is heated to approximately 170 ºC, but the crystallinity significantly increases, suggesting that the exothermic signal is due to amorphous transformation. HPLC / IC results indicate that the molar ratio of fumarate crystal form A is 1.07 (acid / base), suggesting that fumarate crystal form A is an amorphous form of fumarate.

[0187] The XRPD characterization results of adipate crystal form A are as follows: Figure 20 TGA results are as follows Figure 21 The TGA curves show that the adipate crystal form A sample lost approximately 2.5% of its weight when heated to about 130 ºC. The DSC results are as follows... Figure 21 The DSC curves show an endothermic peak at approximately 182.0ºC (initial temperature), suggesting that adipate crystal form A is amorphous. HPLC / IC results indicate that the molar ratio of adipate crystal form A is 0.88 (acid / base).

[0188] The XRPD results of the benzoate crystal form A are as follows: Figure 26 As shown. TGA results are as follows. Figure 27 The TGA curves show that benzoate crystal form A loses approximately 2.7% of its weight when heated to about 130ºC. The DSC results are as follows... Figure 27The DSC curves show an endothermic peak at approximately 169.5ºC (initial temperature), suggesting that benzoate crystal form A is an amorphous form. HPLC / IC results indicate that the molar ratio of benzoate crystal form A is 0.81 (acid / base).

[0189] The XRPD results of the hydrochloride crystal form A are as follows: Figure 7 As shown in the figure. TGA results are as follows. Figure 8 The TGA curve in the image shows that the sample loses approximately 3.5% of its weight when heated to about 100 °C. The DSC results are as follows... Figure 8 The DSC curves show multiple endothermic and exothermic peaks, specifically endothermic peaks with peak temperatures of approximately 94.6°C, 237.4°C, and 266.9°C, and exothermic peaks with peak temperatures of approximately 156.2°C and 203.0°C, respectively. Furthermore, the endothermic peaks with peak temperatures of approximately 237.4°C and 266.9°C have onset temperatures of approximately 233.7°C and 261.9°C, respectively.

[0190] The XRPD results of the hydrochloride crystal form B sample are as follows: Figure 7 As shown in the figure. TGA results are as follows. Figure 9 The TGA curve in the image shows that the sample loses approximately 9.1% of its weight when heated to about 130 °C. The DSC results are as follows: Figure 9 The DSC curves show endothermic peaks at approximately 109.9 ºC, approximately 160.3 ºC, and approximately 266.9 ºC (peak temperature).

[0191] The XRPD results of the phosphate crystal form A are as follows: Figure 10 As shown. TGA results are as follows. Figure 11 The TGA curves show that the phosphate crystal form A sample loses approximately 3.6% of its weight when heated to about 130ºC. The DSC results are as follows: Figure 11 The DSC curves show endothermic peaks at approximately 48.0ºC and approximately 228.3ºC (starting temperature).

[0192] The XRPD results of lactate crystal form A are as follows: Figure 12 As shown. TGA results are as follows. Figure 13 The TGA curves show that the lactate crystal form A sample loses approximately 4.0% of its weight when heated to about 130ºC. The DSC results are as follows: Figure 13 The DSC curves show endothermic peaks at approximately 99.0ºC and approximately 183.3ºC (peak temperature).

[0193] The XRPD results of the succinate crystal form A are as follows: Figure 16 As shown. TGA results are as follows. Figure 17The TGA curves show that the succinate crystal form A sample lost approximately 3.2% of its weight when heated to about 130 ºC. The DSC results are as follows... Figure 17 The DSC curves show endothermic peaks at approximately 51.6ºC and approximately 182.1ºC (starting temperature). Heating tests show that the crystal form of the sample remains unchanged when heated to 100ºC.

[0194] The XRPD results of the malate crystal form A are as follows: Figure 18 As shown. TGA results are as follows. Figure 19 The TGA curves show that the malate crystal form A sample loses approximately 4.0% of its weight when heated to about 130 °C. The DSC results are as follows: Figure 19 The DSC curves show that there are multiple endothermic peaks before decomposition.

[0195] The XRPD characterization results of hippurate crystal form A are as follows: Figure 22 TGA results are as follows Figure 23 The TGA curves show that the hippurate crystal form A sample lost approximately 4.2% of its weight when heated to about 130 ºC. The DSC results are as follows: Figure 23 The DSC curves show that there are multiple endothermic peaks before decomposition, especially the endothermic peaks with peak temperatures of approximately 124.2°C, approximately 141.0°C, approximately 154.6°C, approximately 178.0°C and approximately 217.2°C.

[0196] The XRPD results of the glycolate crystal form A are as follows: Figure 24 TGA results are as follows Figure 25 The TGA curves show that the glycolate crystal form A sample loses approximately 4.8% of its weight when heated to about 130 °C. The DSC results are as follows: Figure 25 The DSC curves show endothermic peaks at approximately 54.1°C and approximately 183.2°C (starting temperature). Heating tests show that the crystal form of the sample remains unchanged when heated to 130°C.

[0197] The XRPD results of the nicotinate crystal form A are as follows: Figure 28 TGA results are as follows Figure 29 The TGA curves show that the nicotinate crystal form A sample loses approximately 4.9% of its weight when heated to about 130 °C. The DSC results are as follows: Figure 29 The DSC curves show that there are multiple endothermic peaks before decomposition, especially the endothermic peaks with peak temperatures of approximately 103.9°C, approximately 160.3°C and approximately 212.5°C.

[0198] Taking into account 1) the safety level of the selected acid (Level I > Level II > Level III); 2) sharp diffraction peaks in the XRPD pattern (at the same time, to ensure the representativeness of the sample and the collected data, those without obvious amorphous peaks are preferred); 3) small and gradual weight loss in TGA; 4) single and relatively sharp melting peaks in DSC (those without crystal form and with low adsorption water or solvent content are preferred), maleate crystal form A, fumarate crystal form A, adipate crystal form A, and benzoate crystal form A are found to be preferred.

[0199] Experimental Example 1 The solubility, solid-state stability, and hygroscopicity of the samples in the 37 ºC state were evaluated for maleate crystal form A, fumarate crystal form A, adipate crystal form A, and benzoate crystal form A, respectively.

[0200] Dynamic solubility at 37°C Dynamic solubility tests were conducted in four solvents: water, simulated gastric juice (SGF), simulated fasting intestinal fluid (FaSSIF), and simulated feeding intestinal fluid (FeSSIF) to assess the solubility and disproportionation risk of maleate crystal form A, fumarate crystal form A, adipate crystal form A, and benzoate crystal form A.

[0201] In the experiment, using free alkali crystal form A as a reference, approximately 32 mg of solid was mixed with 4.0 mL of solvent in a 5 mL glass bottle, sealed, and fixed on a rotating disk at 25 rpm. Samples were taken after mixing at 37ºC for 1 hour, 2 hours, 4 hours, and 24 hours. Turbid samples were centrifuged, and the supernatant was used to determine the HPLC concentration and pH. The crystal form of the solid was determined using XRPD. If the sample remained clear, a sample was taken after 24 hours to test the concentration and pH.

[0202] HPLC data were collected on an Agilent 1100 HPLC system. The solubility test parameters are shown in Table 9 below.

[0203] Table 9 Solubility Test Parameters

[0204] The results of dynamic solubility tests of the four salt crystal forms A and free alkali crystal form A at 37 ºC are summarized in Table 10.

[0205] Table 10: Summary of Dynamic Solubility Test Results

[0206] Note: S 1 Solubility, μg / mL; measured data is free alkali concentration; pH 2Measured pH and FC values ​​at each sampling point 3 : Changes in crystal form after XRPD detection. --: No data collected from sample dissolution. NA: Undetectable due to small sample volume.

[0207] Free base: Free base crystal form A, Maleate: Maleate crystal form A, Fumarate: Fumarate crystal form A, Adipate: Adipate crystal form A, Benzoate: Benzoate crystal form A.

[0208] The results showed that: 1) In water, the solubility of crystal form A of the four salts was significantly higher than that of free alkali crystal form A (from a few micrograms to a few milligrams per milliliter), while the crystal form of maleate crystal form A remained unchanged after being suspended in water for 24 hours; 2) In other solvents, the solubility of crystal form A of the four salts was similar to that of free alkali crystal form A, both being a few milligrams per milliliter. However, oil formation was observed in all samples after suspension in FeSSIF for 1 hour. Compared with the observations in FaSSIF, the in vitro solubility test results suggest that food effects may need to be considered for optimized absorption.

[0209] One-week SSD stability Appropriate amounts of samples were weighed and placed in the open at 25 ºC / 60%RH and 40 ºC / 75%RH respectively. After one week, all samples were characterized by XRPD and tested by HPLC to detect changes in crystal form and purity.

[0210] The results of the one-week stability assessment for maleate crystal form A, fumarate crystal form A, adipate crystal form A, benzoate crystal form A and free base crystal form A are summarized in Table 11.

[0211] Table 11

[0212] The results showed that no changes in crystal form or HPLC purity occurred in the four salt forms and the free base at 25 ºC / 60%RH, demonstrating good physicochemical stability; however, the HPLC purity of adipate crystal form A and benzoate crystal form A samples decreased after storage at 40 ºC / 75%RH.

[0213] Hygroscopic To assess the stability of samples under 25 °C with varying humidity, DVS was used to test maleate crystal form A, fumarate crystal form A, adipate crystal form A, and benzoate crystal form A. All samples were pre-dried at 0% RH to remove adsorbed solvent or water before testing.

[0214] The following conclusions can be drawn from the test results: 1) At 80% RH, the moisture adsorption (wt%) of maleate crystal form A and fumarate crystal form A were 0.8% and 1.0%, respectively, indicating slight hygroscopicity; while the moisture adsorption of adipate crystal form A and benzoate crystal form A were 9.1% and 7.7%, respectively, also indicating hygroscopicity. Furthermore, XRPD tests were performed before and after the DVS test. The results showed that the crystal forms of maleate crystal form A and adipate crystal form A remained unchanged, while the crystallinity of fumarate crystal form A and benzoate crystal form A decreased.

[0215] 2) At 80%RH to 95%RH, except for maleate crystal form A which absorbs less water, the water absorption of the other three salt forms increased rapidly.

[0216] In summary, the solubility of maleate crystal form A, fumarate crystal form A, adipate crystal form A, and benzoate crystal form A in water and biological solvents is similar to or higher than that of free base crystal form A. Except for adipate and benzoate, which showed a decrease in HPLC purity, the other salt forms and free base crystal form A exhibited good physicochemical stability in the solid-state stability assessment. According to the water adsorption results, maleate crystal form A and fumarate crystal form A samples have slight hygroscopicity, while the other salt forms and free base crystal form A all showed hygroscopicity.

[0217] Based on the above evaluations of dynamic solubility, solid stability, and hygroscopicity, maleate crystal form A exhibits superior properties, primarily in the following ways: 1) Under the same operating conditions, the laboratory batch of samples had higher crystallinity, while the fumarate crystal form A, adipate crystal form A and benzoate crystal form A had moderate crystallinity; 2) Compared with free alkali crystal form A, maleate crystal form A showed a significant increase in water solubility at 37 ºC (from a few micrograms to a few milligrams / mL), and the crystal form did not change after 24 hours; the solubility in other solvents (simulated gastric juice SGF, simulated fasting intestinal juice FaSSIF, and simulated feeding intestinal juice FeSSIF) was similar; 3) Maleate crystal form A sample did not change in crystal form or HPLC purity after being placed at 25 ºC / 60%RH and 40 ºC / 75%RH for one week, demonstrating good physicochemical stability; 4) Dynamic moisture adsorption (DVS) test results at 25 °C showed that maleate crystal form A sample has slight hygroscopicity and its moisture adsorption capacity is lower than that of other candidate salts and free bases under high humidity conditions (95%RH).

[0218] Preparation Example 2 Crystal forms are prepared using the following general methods: Weigh approximately 200 mg of free base and add the corresponding acid and 5 mL of EtOAc in a 1:1 molar ratio to obtain a suspension. Stir at room temperature for 30 minutes. Add ~2 mg of seed crystals and magnetically stir overnight under temperature cycling (50-20 ºC, 0.1 ºC / min, 3 times; rotation speed 500 rpm). Centrifuge to separate the solid, vacuum dry at room temperature for 5 hours, collect the crystalline sample, and perform XRPD and DSC tests.

[0219] Maleate, fumarate, adipate, and benzoate crystal forms were prepared using the above method. XRPD and DSC test results showed that they were consistent with the references maleate crystal form A, fumarate crystal form A, adipate crystal form A, and benzoate crystal form A, respectively.

[0220] Preparation Example 3 3.1 Preparation of maleate crystal form B Add the antisolvent to a clear solution of maleate of compound I in the normal solvent while stirring dropwise (~1000 rpm) until solid precipitates. Stop the process if no solid precipitates after adding approximately 10 mL of antisolvent. Centrifuge (~10000 rpm, 2 min) to separate the precipitated solid and perform XRPD analysis.

[0221] The results are shown in Table 12, which yielded maleate crystal forms A, B, amorphous and oil.

[0222] Table 12

[0223] In addition, maleate crystal form A of compound I was suspended in CHCl3 / n-hexane (1:1, v / v) and magnetically stirred at 50°C (~1000 rpm) for about 4 days. The solid was then collected by centrifugation (~10000 rpm, 3 min) and subjected to XRPD testing. Maleate crystal form B was also obtained.

[0224] The XRPD results for maleate crystal form B are as follows: Figure 3 The TGA characterization results are as follows: Figure 4 The TGA curves show that the sample lost 6.4% of its weight when heated to approximately 200 °C (the weight loss of approximately 19.1% between 200 and 290 °C is presumably due to deacidification, with a theoretical weight loss of approximately 18.7% for deacidification). DSC characterization results are as follows... Figure 4 The DSC curves show two weak endothermic peaks at approximately 74.6 °C and approximately 236.8 °C (peak temperature), and a sharp endothermic peak at approximately 225.5 °C (starting temperature). 1 ¹H NMR results showed that the molar ratio of maleic acid to free base was 1:1.

[0225] 3.2 Preparation of maleate crystal form C Maleate crystal form B is transformed into maleate crystal form C after being purged with N2 at 30 °C for 20 min.

[0226] The XRPD results for maleate crystal form C are as follows: Figure 5 The TGA characterization results are as follows: Figure 6 The TGA curves show that the sample loses approximately 6.3% of its weight when heated to about 200 °C (the weight loss of approximately 17.5% between 200 and 290 °C is presumably due to deacidification, with a theoretical weight loss of approximately 18.7% for deacidification). DSC characterization results are as follows... Figure 6 The DSC curves show an exothermic peak at approximately 142.8 °C (peak temperature) and a sharp endothermic peak at approximately 222.0 °C (starting temperature). 1 ¹H NMR results showed that the molar ratio of maleic acid to free base was 1:1. VT-XRPD results showed that the crystal form of maleate C remained unchanged when heated to approximately 110 °C under N₂ protection. Based on these results, it is speculated that maleate crystal form C is an amorphous form.

[0227] Maleate crystal form C transforms into maleate crystal form A when heated to approximately 170 ºC under N2 protection.

[0228] Based on the above results, it is speculated that maleate crystal form B is a hydrate, which is transformed into an amorphous form after being purged with N2 to remove the bound water.

[0229] 3.3 Slow Volatilization Preparation Experiment Seal a vial containing a clear solution of maleate of compound I in one of the solvents listed in Table 13 below with sealing film and poke five small holes in it, and allow it to evaporate slowly at room temperature. Once the solvent has completely evaporated, collect the resulting solid and perform an XRPD test.

[0230] The experimental results are shown in Table 13. Crystal form A and amorphous form were obtained in the slow volatilization test.

[0231] Table 13

[0232] 3.4 Slow Cooling Preparation Experiment The clear solution of maleate containing compound I in the solvents listed in Table 14 below was placed in a biochemical incubator and cooled from 50 °C to 5 °C at a cooling rate of 0.1 °C / min. The unprecipitated solid was transferred to room temperature to evaporate, and the resulting solid was collected and subjected to XRPD testing.

[0233] The experimental results are shown in Table 14. The slow cooling test yielded an amorphous product.

[0234] Table 14

[0235] Note: *: Obtained by vacuum drying at room temperature.

[0236] Experimental Example 2 1. Hybrid suspension competition test To further investigate the stability relationship among the amorphous forms A and C and the hydrated form B of maleate, a suspension competition experiment was conducted among the various crystal forms. Specifically, this included the suspension competition of crystal forms A and C in acetone and EtOAc at room temperature and 50 °C, and the suspension competition between amorphous form A and hydrated form B in MeOH / H₂O (water activity a) at room temperature. w The suspension competition test in (=0~1).

[0237] The specific steps are as follows: 1) Prepare saturated solutions of maleate amorphous form A in different solvents at a specific temperature; 2) Add equal masses of crystal form A and B or C samples (approximately 4 mg each) to 0.5 mL of the saturated solution to form suspensions; 3) Suspend and stir for approximately 5 days (~800 rpm) at room temperature and 50 °C respectively; 4) Separate the remaining solids and test XRPD.

[0238] The results of the hybrid suspension competition test are summarized in Table 15.

[0239] Table 15

[0240] The results showed that at room temperature and 50 °C, a physical mixture of crystal forms A and C transformed into crystal form A after suspension and stirring in acetone and EtOAc; at room temperature, a physical mixture of crystal forms A and B transformed into crystal form A in MeOH / H2O (a w After being suspended and stirred in a solution of 0~1), it transforms into crystal form A. Based on this, it is inferred that the water activity a at room temperature to 50 °C... w When the saturation value is 0~1, crystal form A is a thermodynamically stable crystal form.

[0241] 2. Equilibrium solubility test The equilibrium solubility of maleate crystal forms A, B, and C in water at room temperature was tested.

[0242] Suspensions of various crystal forms were prepared in the experiment, and after being magnetically stirred at room temperature for 24 hours (speed ~800 rpm), they were centrifuged (10000 rpm, 5 min). The supernatant was filtered (using a 0.22 μm PTFE filter membrane) and its solubility and pH were measured. XRPD of the solid was measured.

[0243] Table 16 summarizes the equilibrium solubility results of maleate crystal forms A, B, and C in water.

[0244] Table 16

[0245] Notes: *: Calculated based on free base equivalent. NA: Remaining solids were low and not tested.

[0246] 3. Solid-state stability test This experiment was used to evaluate the solid-state stability of maleate crystal form A.

[0247] Weigh out an appropriate amount of maleate crystal form A and place it in a closed container at 80 °C for 1 day, and then in an open container at 25 °C / 60%RH and 40 °C / 75%RH for 1 week. The solid samples separated under different conditions were subjected to HPLC to assess purity and XRPD to assess crystal form to assess physical stability.

[0248] The results of the solid-state stability assessment of maleate crystal form A are summarized in Table 17.

[0249] Table 17

[0250] No crystal form transformation or decrease in HPLC purity occurred under any of the three test conditions, indicating that maleate crystal form A has good physical and chemical stability under the selected test conditions.

[0251] 4. Stability in solvents This experiment was used to evaluate the stability of the maleate salt of compound I in solvents.

[0252] The stability of compound I maleate in solvents was evaluated by suspension stirring tests in dichloromethane (DCM), methanol (MeOH), ethyl acetate (EtOAc), dimethylformamide (DMF), tetrahydrofuran (THF), MTBE, and 1,4-dioxane.

[0253] The specific steps are as follows: Weigh ~15 mg of maleate crystal form A of compound I per sample, add it to different solvents, suspend and stir at room temperature for 6 h, then transfer to 60 ºC and stir for 5 h, centrifuge to separate the solid and measure XRPD, and use the solution for HPLC purity analysis.

[0254] The results showed that maleate crystal form A remained unchanged after suspension and stirring in DCM, EtOAc, THF, and MTBE, and its purity did not decrease significantly; after suspension and stirring in MeOH and DMF, its crystal form remained unchanged, and its purity increased slightly; after suspension and stirring in 1,4-dioxane, its crystal form remained unchanged, and its purity decreased slightly.

[0255] The results of the stability assessment of maleate crystal form A in solvents are summarized in Tables 18 and 19.

[0256] Table 18

[0257] Table 19

[0258] Experimental Example 3: Comparison of the stability of free alkali crystal form A and maleate crystal form A The free alkali crystal form A and the maleate crystal form A obtained therefrom were subjected to high temperature (60℃) and light irradiation (1.2*10⁻⁶) tests. 6 Lux*hr) conducted a 5-day investigation, and the testing items were related substances.

[0259] Related substance analysis methods for free base Instrument: High Performance Liquid Chromatography (HPLC) Chromatographic column: Waters XBridge C18, 150*4.6mm, 3.5µm Mobile phase A: 10 mmol ammonium dihydrogen phosphate (containing 0.1% triethylamine): methanol (80:20) Mobile phase B: Methanol Gradient table:

[0260] Detection wavelength: 230nm Flow rate: 1.0 mL / min Injection volume: 20 μl Column temperature: 45℃ Runtime: 45 min Test solution: Take about 25 mg of this product, place it in a 100 ml volumetric flask, dissolve and dilute with methanol to the mark, shake well, and use it as the test solution.

[0261] Related material analysis methods for maleate crystal form A Instrument: High Performance Liquid Chromatography (HPLC) Column: Waters Xbridge C18, 4.6 × 150 mm, 3.5 μm Mobile phase A: 0.01M ammonium dihydrogen phosphate aqueous solution (containing 0.1% triethylamine): methanol = 80:20 Mobile phase B: Methanol Gradient table:

[0262] Detection wavelength: 230nm Flow rate: 1.0 ml / min Injection volume: 20 μl Column temperature: 45℃ Runtime: 45 min Diluent: 0.01M ammonium dihydrogen phosphate aqueous solution (containing 0.1% triethylamine): methanol = 30:70 Test solution: Take about 30 mg of this product, place it in a 100 ml volumetric flask, add diluent to dissolve and dilute to the mark, shake well, and use it as the test solution.

[0263] The data obtained is as follows: High-temperature and light-related material data for free alkali crystal form A / maleate crystal form A

[0264] The table above shows that free alkali is more effective at high temperatures (60℃) and under light (1.2*10). 6 In a 5-day experiment, the Lux·hr form showed instability. However, the maleate form was more stable than the free base form under the same conditions.

Claims

1. The maleate crystal form A of compound I is shown below: in, The X-ray powder diffraction pattern of the maleate crystal form A has characteristic peaks at diffraction angles 2θ of 5.48±0.2°, 8.55±0.2°, 10.92±0.2°, 12.04±0.2°, 12.98±0.2°, 13.81±0.2°, 16.40±0.2°, 19.59±0.2°, and 27.46±0.2°.

2. The maleate crystal form A of compound I according to claim 1, wherein, The X-ray powder diffraction pattern of the maleate crystal form A shows diffraction peaks at one or more locations at diffraction angles of 2θ, namely 6.11 ±0.2°, 7.84 ±0.2°, 15.32 ±0.2°, 18.70 ±0.2°, 21.55 ±0.2°, 22.21 ±0.2°, 22.80 ±0.2°, 23.32 ±0.2°, 24.57 ±0.2°, 25.62 ±0.2°, 26.07 ±0.2°, 28.23 ±0.2°, 28.68 ±0.2°, 33.08 ±0.2°, and 38.76 ±0.2°.

3. The maleate crystal form A of compound I according to claim 1, wherein, The X-ray powder diffraction patterns of the maleate crystal form A at diffraction angles 2θ were 5.48±0.2°, 6.11±0.2°, 7.84±0.2°, 8.55±0.2°, 10.92±0.2°, 12.04±0.2°, 12.98±0.2°, 13.81±0.2°, 15.32±0.2°, 16.40±0.2°, 18.70±0.2°, 19.59±0.2°, 21.55±0.2°, 22.21±0.2°, 22.80±0.2°, 23.32±0.2°, 24.57±0.2°, 25.62±0.2°, 26.07±0.2°, 27.46±0.2°, and 28.

23. Characteristic peaks are present at ±0.2°, 28.68±0.2°, 33.08±0.2° and 38.76±0.2°.

4. A method for preparing maleate crystal form A according to any one of claims 1-3, wherein, The compound I is prepared by reacting its free base with maleic acid in a solvent selected from acetone, tetrahydrofuran, and ethyl acetate.

5. A pharmaceutical composition comprising maleate crystal form A selected from compound I according to any one of claims 1-3.

6. Use of the maleate crystal form A of compound I according to any one of claims 1-3 or the pharmaceutical composition according to claim 5 for the preparation of a medicament for the prevention or treatment of diseases associated with abnormalities of cyclin-dependent kinases CDK4 and / or CDK6.

7. The use according to claim 6, wherein the disease is a tumor.

8. The use according to claim 7, wherein, The tumor is selected from the following malignant tumors: breast cancer, colon cancer, astrocytoma, chronic myeloid leukemia, pancreatic cancer, acute monocytic leukemia, liver cancer, gastric cancer, non-small cell lung cancer, glioblastoma, and prostate adenocarcinoma.

9. Use of the maleate crystal form A of compound I according to any one of claims 1-3 or the pharmaceutical composition according to claim 5 for the preparation of cyclin-dependent kinase CDK4 and / or CDK6 inhibitors.

10. Use of the maleate crystal form A of compound I according to any one of claims 1-3 or the pharmaceutical composition according to claim 5 for the preparation of a medicament for inhibiting the proliferation of tumor cells, wherein the tumor cells are cancer cells.

11. The use according to claim 10, wherein, The cancer cells are selected from breast cancer cells, colon cancer cells, brain astrocytoma cells, chronic myeloid leukemia cells, pancreatic cancer cells, acute monocytic leukemia cells, liver cancer cells, gastric cancer cells, non-small cell lung cancer cells, malignant glioblastoma cells, and prostate adenocarcinoma cells.

12. The use according to claim 11, wherein, The breast cancer cells are selected from one or more of the following breast cancer cells: MCF-7, T-47D, and ZR-75-1.

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