Pharmaceutically acceptable salts and polymorphs of substituted pyrazolo [1, 5-a] pyrimidine-7-amine derivatives and uses thereof

Abstract

Relates to a pharmaceutically acceptable salt and a polymorphic substance of a substituted pyrazolo [1, 5-a] pyrimidine-7-amine derivative as shown in a formula (I), a pharmaceutical composition containing the same, a preparation method of the pharmaceutical composition, and application of the pharmaceutically acceptable salt and the polymorphic substance in treatment or prevention of CDK9 activity-related diseases or CDK9 activity-mediated diseases. # imgabs0 #

Description

Pharmaceutically acceptable salts and polymorphs of substituted pyrazolo[1,5-a]pyrimidin-7-amine derivatives and their uses

[0001] Related applications

[0002] This application claims priority to Chinese patent applications filed with the China Patent Office on July 22, 2022, with application number No. 202210866135.0, entitled “Pharmaceutically acceptable salts and polymorphs of substituted pyrazolo[1,5-a]pyrimidine-7-amine derivatives and their applications” and application number No. 202210870337.2, entitled “Phosphate salts and polymorphs of CDK9 inhibitors and their applications”, the entire text of which is hereby incorporated by reference. Technical Field

[0003] The present disclosure belongs to the field of medical technology, and specifically relates to pharmaceutically acceptable salts and polymorphs of substituted pyrazolo[1,5-a]pyrimidin-7-amine derivatives, pharmaceutical compositions containing the same, preparation methods thereof, and medical applications thereof. Background Art

[0004] CDK9 is a member of the cyclin-dependent kinase (CDK) protein family and plays an important role in gene transcription regulation. CDK9 mainly regulates gene transcription elongation by phosphorylating the carbon-terminal region of RNA complex II. CDK9 is widely and highly expressed in tumors and is also an important factor in the progression and maintenance of tumor cells. CDK9 inhibitors downregulate the expression of related oncoproteins (MYC) and the apoptosis inhibitor protein Mcl-1 by inhibiting gene transcription elongation, thereby promoting the apoptosis of cancer cells. CDK9 inhibitors reactivate silent genes by regulating the epigenetic factor BRG1, including the activation of endogenous retroviruses (ERVs) in tumor cells, promoting interferon expression, and making tumor cells more sensitive to immunotherapy.

[0005] Currently, many companies are developing CDK9 inhibitors, including the selective CDK9 inhibitor BAY1251152 developed by Bayer, the selective CDK9 inhibitor AZD4573 developed by AstraZeneca, the non-selective CDK9 inhibitor TP-1287 developed by Tolero, and the non-selective CDK9 inhibitor QHRD107 developed by Changzhou Qianhong Pharmaceutical.

[0006] The solubility and stability of active ingredients in pharmaceuticals have an important impact on drugability research, and the solid form is beneficial to product quality control. Therefore, it is necessary to explore the solid form of the product.

[0007] Summary of the Invention

[0008] Based on this, the purpose of the present disclosure is to provide pharmaceutically acceptable salts and polymorphs of substituted pyrazolo[1,5-a]pyrimidin-7-amine derivatives and their uses. Specifically, the pharmaceutically acceptable salts are L-tartrate and phosphate salts of substituted pyrazolo[1,5-a]pyrimidin-7-amine derivatives. The substituted pyrazolo[1,5-a]pyrimidin-7-amine derivative is a CDK9 inhibitor, and its molecular structure is (1S,3S)-N1-(5-((S)-1-cyclobutylethyl)pyrazolo[1,5-a]pyrimidin-7-yl)cyclopentane-1,3-diamine shown in formula (I).

[0009] In a first aspect of the present disclosure, there is provided a pharmaceutically acceptable salt of a compound of formula (I):

[0010] Wherein, the pharmaceutically acceptable salt is selected from: phosphate and L-tartrate.

[0011] In some embodiments, the pharmaceutically acceptable salt of the compound of formula (I) is in an anhydrous form, a hydrate form, or a solvate form.

[0012] In some embodiments, the pharmaceutically acceptable salt of the compound of formula (I) is crystalline. In some embodiments, the pharmaceutically acceptable salt of the compound of formula (I) is amorphous.

[0013] In some embodiments, the pharmaceutically acceptable salt is a phosphate salt. In some embodiments, the phosphate salt is a first-class phosphate salt of a compound of formula (I), wherein the molar ratio of phosphoric acid to the compound of formula (I) is (1.8-2.4):1. In some embodiments, the molar ratio of phosphoric acid to the compound of formula (I) can be 1.8:1, 1.9:1, 2.0:1, 2.1:1, 2.2:1, 2.3:1, or 2.4:1. In some embodiments, the molar ratio of phosphoric acid to the compound of formula (I) is (1.9-2.3):1. In one embodiment, the molar ratio of phosphoric acid to the compound of formula (I) is 2:1.

[0014] In some embodiments, the phosphate is a phosphate salt of the compound of formula (I) in Form I, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following diffraction angles 2θ (°): 18.234±0.2, 19.131±0.2, and 21.266±0.2. In some embodiments, the phosphate is a phosphate salt of the compound of formula (I) in Form I, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following diffraction angles 2θ (°): 18.803±0.2, 18.234±0.2, 19.131±0.2, and 21.266±0.2.

[0015] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystalline form I has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 18.234 ± 0.2, 19.131 ± 0.2 and 21.266 ± 0.2, and further includes characteristic diffraction peaks at 2 or more (such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.) selected from the following group of diffraction angle 2θ (°) values: 2.121 ± 0.2, 7.373 ± 0.2, 9.556 ± 0.2, 10.607 ± 0.2, 1 1.105±0.2, 12.322±0.2, 12.917±0.2, 13.999±0.2, 14.902±0.2, 15.238±0.2, 18.803±0.2, 19.881±0.2, 20.697±0.2, 22.01±0.2, 22.464±0.2, 23.247±0.2, 24.025±0.2, 26.367±0.2, 28.513±0.2, 30.297±0.2, 31.216±0.2 and 34.017±0.2.

[0016] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystalline form I has characteristic diffraction peaks at 3 or more (such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.) or all selected from the following diffraction angle 2θ (°) values: 2.121 ± 0.2, 7.373 ± 0.2, 9.556 ± 0.2, 10.607 ± 0.2, 11.105 ± 0.2, 12.322 ± 0.2, 12.917 ± 0.2, 13.999 ± 0.2, 14.9 02±0.2, 15.238±0.2, 18.234±0.2, 18.803±0.2, 19.131±0.2, 19.881±0.2, 20.697±0.2, 21.266±0.2, 22.01±0.2, 22.464±0.2, 23.247±0.2, 24.025±0.2, 26.367±0.2, 28.513±0.2, 30.297±0.2, 31.216±0.2 and 34.017±0.2.

[0017] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystalline form I has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 13.999±0.2, 18.234±0.2, 18.803±0.2, 19.131±0.2, 21.266±0.2, 22.01±0.2 and 23.247±0.2.

[0018] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystalline form I has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 7.373±0.2, 10.607±0.2, 11.105±0.2, 12.322±0.2, 12.917±0.2, 13.999±0.2, 14.902±0.2, 15.238±0.2, 18.234±0.2, 18.803±0.2, 19.131±0.2, 20.697±0.2, 21.266±0.2, 22.01±0.2, 22.464±0.2, 23.247±0.2, 24.025±0.2, 26.367±0.2 and 28.513±0.2.

[0019] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystal form I has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 2.121±0.2, 7.373±0.2, 9.556±0.2, 10.607±0.2, 11.105±0.2, 12.322±0.2, 12.917±0.2, 13.999±0.2, 14.902±0.2, 15.238±0.2, 18.23 4±0.2, 18.803±0.2, 19.131±0.2, 19.881±0.2, 20.697±0.2, 21.266±0.2, 22.01±0.2, 22.464±0.2, 23.247±0.2, 24.025±0.2, 26.367±0.2, 28.513±0.2, 30.297±0.2, 31.216±0.2 and 34.017±0.2.

[0020] In some embodiments, the X-ray powder diffraction pattern of the phosphate salt of the compound of formula (I) in Form I has characteristic diffraction peaks expressed in 2θ (°) values ​​and d values ​​as shown in Table 1, and the relative intensity of each peak is shown in Table 1.

[0021] Table 1. 2θ(°), d value and relative intensity I / I0 of phosphate crystal form I

[0022] In some embodiments, the molar ratio of phosphoric acid to the compound of formula (I) in the phosphate salt crystal form I is 2:1.

[0023] In some embodiments, the X-ray powder diffraction pattern of Form I of the phosphate salt of the compound of formula (I) is substantially as shown in FIG1 .

[0024] In some embodiments, the differential scanning calorimetry curve of the phosphate salt of the compound of formula (I) Form I has an endothermic peak at 188.01°C ± 3°C, 188.01°C ± 2°C, 188.01°C ± 1°C, or 188.01°C ± 0.5°C. In some embodiments, the onset temperature in the differential scanning calorimetry curve of the phosphate salt of the compound of formula (I) Form I is 188.01°C ± 3°C, 188.01°C ± 2°C, 188.01°C ± 1°C, or 188.01°C ± 0.5°C, and the peak temperature is 193.69°C ± 3°C, 193.69°C ± 2°C, 193.69°C ± 1°C, or 193.69°C ± 0.5°C. In some embodiments, the differential scanning calorimetry analysis spectrum (DSC spectrum) of the phosphate salt of the compound of formula (I) Form I is substantially as shown in Figure 2. In the embodiment shown in FIG2 , the melting point of the phosphate crystal form I of the compound of formula (I) is approximately 188.01±0.5° C.

[0025] In some embodiments, the thermogravimetric analysis (TGA) pattern of the phosphate salt of the compound of formula (I) Form I is substantially as shown in Figure 2. In some embodiments, the TGA pattern of the phosphate salt of the compound of formula (I) Form I shows a weight loss of 1.139% at around 190°C.

[0026] In some embodiments, the dynamic moisture absorption spectrum (DVS graph) of the phosphate salt of the compound of formula (I) in Form I is substantially as shown in Figure 3. In some embodiments, the DVS graph of the phosphate salt of the compound of formula (I) in Form I shows that under 80% relative humidity (RH), the weight gain due to moisture absorption is 9%.

[0027] In some embodiments, a microscopic image of Form I of the phosphate salt of the compound of formula (I) is substantially as shown in Figure 4. In some embodiments, a microscopic image of Form I of the phosphate salt of the compound of formula (I) shows that Form I of the phosphate salt is in the form of rods and blocks.

[0028] In this first aspect, in some embodiments, the pharmaceutically acceptable salt is an L-tartrate salt. The molar ratio of L-tartaric acid to the compound of formula (I) in the L-tartrate salt is (0.8-1.2):1. In some embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) can be 0.8:1, 0.9:1, 1.0:1, 1.1:1 or 1.2:1. In some embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) is (0.9-1.1):1. In one embodiment, the molar ratio of L-tartaric acid to the compound of formula (I) is 1:1.

[0029] In some embodiments, the L-tartrate salt is L-tartrate salt form I of the compound of formula (I), and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following group of diffraction angles 2θ (°) values: 13.946±0.2, 16.881±0.2, 19.405±0.2, 21.505±0.2 and 24.262±0.2.

[0030] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form I has characteristic diffraction peaks at the following group of diffraction angle 2θ (°) values: 13.946 ± 0.2, 16.881 ± 0.2, 19.405 ± 0.2, 21.505 ± 0.2 and 24.262 ± 0.2, and further includes 2 or more (such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.) selected from the following group of characteristic diffraction peaks at diffraction angle 2θ (°): 6.687 ± 0.2, 7.436 ± 0.2, 9.493 ± 0.2, 10.615±0.2, 12.053±0.2, 12.776±0.2, 13.164±0.2, 14.875±0.2, 15.201±0.2, 16.013±0.2, 18.175±0.2, 19.045±0.2, 20.659±0.2, 22.434±0.2, 23.04±0.2, 25.202±0.2, 26.452±0.2, 28.105±0.2, 29.692±0.2, 31.579±0.2, 34.139±0.2 and 34.543±0.2.

[0031] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form I has characteristic diffraction peaks at 5 or more (such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.) or all selected from the following diffraction angle 2θ (°) values: 6.687±0.2, 7.436±0.2, 9.493±0.2, 10.615±0.2, 12.053±0.2, 12.776±0.2, 13.164±0.2, 13.946±0.2, 14.875±0.2, 15.2 01±0.2, 16.013±0.2, 16.881±0.2, 18.175±0.2, 19.045±0.2, 19.405±0.2, 20.659±0.2, 21.505±0.2, 22.434±0.2, 23.04±0.2, 24.262±0.2, 25.202±0.2, 26.452±0.2, 28.105±0.2, 29.692±0.2, 31.579±0.2, 34.139±0.2 and 34.543±0.2.

[0032] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form I has characteristic diffraction peaks at the following diffraction angles 2θ (°): 7.436±0.2, 13.946±0.2, 16.013±0.2, 16.881±0.2, 18.175±0.2, 19.045±0.2, 19.405±0.2, 21.505±0.2, 24.262±0.2, 25.202±0.2, 26.452±0.2, 28.105±0.2 and 31.579±0.2.

[0033] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form I has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 6.687±0.2, 7.436±0.2, 10.615±0.2, 12.053±0.2, 13.164±0.2, 13.946±0.2, 14.875±0.2, 15.201±0.2, 16.013±0.2, 16.881±0.2, 18. 175±0.2, 19.045±0.2, 19.405±0.2, 20.659±0.2, 21.505±0.2, 22.434±0.2, 23.04±0.2, 24.262±0.2, 25.202±0.2, 26.452±0.2, 28.105±0.2, 29.692±0.2, 31.579±0.2, 34.139±0.2 and 34.543±0.2.

[0034] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form I has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 6.687±0.2, 7.436±0.2, 9.493±0.2, 10.615±0.2, 12.053±0.2, 12.776±0.2, 13.164±0.2, 13.946±0.2, 14.875±0.2, 15.201±0.2, 16.013±0.2. 2, 16.881±0.2, 18.175±0.2, 19.045±0.2, 19.405±0.2, 20.659±0.2, 21.505±0.2, 22.434±0.2, 23.04±0.2, 24.262±0.2, 25.202±0.2, 26.452±0.2, 28.105±0.2, 29.692±0.2, 31.579±0.2, 34.139±0.2 and 34.543±0.2.

[0035] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form I has characteristic diffraction peaks expressed in 2θ (°) values ​​and d values ​​as shown in Table 2, and the relative intensity of each peak is shown in Table 2.

[0036] Table 2. 2θ (°), d value and relative intensity I / I0 of L-tartrate salt form I

[0037] In some embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) in the L-tartrate salt form I is 1:1.

[0038] In some embodiments, the X-ray powder diffraction pattern of Form I of the L-tartrate salt of the compound of formula (I) is substantially as shown in FIG5 .

[0039] In some embodiments, the thermogravimetric analysis spectrum of the L-tartrate salt of the compound of formula (I), Form I, is substantially as shown in Figure 6. In some embodiments, the TGA spectrum of the L-tartrate salt of the compound of formula (I), Form I, shows a weight loss of 1.057% at around 110°C and a further weight loss of 3.482% at around 178°C.

[0040] In some embodiments, the L-tartrate salt is L-tartrate salt form II of the compound of formula (I), and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following set of diffraction angles 2θ (°): 11.662±0.2 and 21.353±0.2.

[0041] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form II has characteristic diffraction peaks at the following group of diffraction angle 2θ (°) values: 11.662 ± 0.2 and 21.353 ± 0.2, and further includes 2 or more (such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, etc.) selected from the following group of characteristic diffraction peaks at diffraction angle 2θ (°): 6.985 ±0.2, 10.376±0.2, 10.884±0.2, 12.994±0.2, 14.244±0.2, 16.548±0.2, 17.481±0.2, 18.349±0.2, 18.984±0.2, 21.026±0.2, 26.925±0.2, 29.335±0.2, 30.896±0.2 and 31.784±0.2.

[0042] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form II has characteristic diffraction peaks at 3 or more (such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.) or all selected from the following diffraction angle 2θ (°) values: 6.985 ± 0.2, 10.376 ± 0.2, 10.884 ± 0.2, 11.662 ± 0.2, 12.994±0.2, 14.244±0.2, 16.548±0.2, 17.481±0.2, 18.349±0.2, 18.984±0.2, 21.026±0.2, 21.353±0.2, 26.925±0.2, 29.335±0.2, 30.896±0.2 and 31.784±0.2.

[0043] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form II has characteristic diffraction peaks at the following diffraction angles 2θ (°): 11.662±0.2, 14.244±0.2, 17.481±0.2, 18.349±0.2, 21.026±0.2 and 21.353±0.2.

[0044] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form II has characteristic diffraction peaks at the following diffraction angles 2θ (°): 10.376±0.2, 11.662±0.2, 14.244±0.2, 16.548±0.2, 17.481±0.2, 18.349±0.2, 18.984±0.2, 21.026±0.2, 21.353±0.2, 26.925±0.2, 29.335±0.2 and 31.784±0.2.

[0045] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form II has characteristic diffraction peaks at the following diffraction angles 2θ (°): 6.985±0.2, 10.376±0.2, 10.884±0.2, 11.662±0.2, 12.994±0.2, 14.244±0.2, 16.548±0.2, 17.481±0.2, 18.349±0.2, 18.984±0.2, 21.026±0.2, 21.353±0.2, 26.925±0.2, 29.335±0.2, 30.896±0.2 and 31.784±0.2.

[0046] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt form II has characteristic diffraction peaks expressed in 2θ (°) values ​​and d values ​​as shown in Table 3, and the relative intensity of each peak is shown in Table 3.

[0047] Table 3. 2θ (°), d value and relative intensity I / I0 of L-tartrate salt form II

[0048] In some embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) in the L-tartrate salt form II is 1:1.

[0049] In some embodiments, the X-ray powder diffraction pattern of the L-tartrate salt Form II is substantially as shown in FIG7 .

[0050] In this first aspect, in some embodiments, the phosphate is a second type phosphate of a compound of formula (I), wherein the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate is 1:(0.8-1.2). In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.9-1.1). In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid can be 1:0.8, 1:0.9, 1:1, 1:1.1, or 1:1.2. In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate is 1:1.

[0051] In some embodiments, the phosphate salt of the compound of formula (I) is in an anhydrous form, a hydrate form, or a solvate form.

[0052] In some embodiments, the phosphate salt of the compound of formula (I) is a crystalline powder and exists in the form of crystals.

[0053] In some embodiments, the phosphate salt of the compound of formula (I) is phosphate crystal form II, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 18.230±0.2 and 21.144±0.2.

[0054] In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate crystal form II is 1:1.

[0055] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystal form II has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 18.230 ± 0.2 and 21.144 ± 0.2, and further includes 2 or more (such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.) selected from the following group of characteristic diffraction peaks at diffraction angle 2θ (°): 10.584 ± 0.2, 11.115 ± 0.2, 11.813 ± 0.2, 12.825 ± 0.2, 13 .969±0.2, 14.873±0.2, 15.313±0.2, 18.771±0.2, 19.85±0.2, 20.576±0.2, 22.01±0.2, 22.492±0.2, 23.153±0.2, 23.96±0.2, 24.947±0.2, 26.273±0.2, 27.837±0.2, 28.432±0.2, 30.143±0.2, 31.072±0.2, 31.88±0.2 and 33.982±0.2.

[0056] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystalline form II has characteristic diffraction peaks at 2 or more (such as 2, 3, 4, 5, 6, 7, 8, 9, 10) or all of the following diffraction angle 2θ (°) values: 10.584±0.2, 13.969±0.2, 14.873±0.2, 18.230±0.2, 20.576±0.2, 21.144±0.2, 22.01±0.2, 22.492±0.2, 23.153±0.2 and 23.96±0.2.

[0057] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystalline form II has characteristic diffraction peaks at the following diffraction angles 2θ (°): 10.584±0.2, 13.969±0.2, 14.873±0.2, 18.230±0.2, 20.576±0.2, 21.144±0.2, 22.01±0.2, 22.492±0.2, 23.153±0.2 and 23.96±0.2.

[0058] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystal form II has characteristic diffraction peaks at 2 or more (such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, etc.) or all selected from the following diffraction angle 2θ (°) values: 10.584±0.2, 11.115±0.2, 11.813±0.2, 12.825±0.2, 13.969±0.2, 14.873±0.2, 15.313±0.2, 18.2 30±0.2, 18.771±0.2, 19.85±0.2, 20.576±0.2, 21.144±0.2, 22.01±0.2, 22.492±0.2, 23.153±0.2, 23.96±0.2, 24.947±0.2, 26.273±0.2, 27.837±0.2, 28.432±0.2, 30.143±0.2, 31.072±0.2, 31.88±0.2 and 33.982±0.2.

[0059] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystal form II has characteristic diffraction peaks at the following diffraction angle 2θ (°) values: 10.584±0.2, 11.115±0.2, 11.813±0.2, 12.825±0.2, 13.969±0.2, 14.873±0.2, 15.313±0.2, 18.230±0.2, 18.771±0.2, 19.85±0.2, 20.576±0.2, 21.144±0.2, 22.01±0.2, 22.492±0.2, 23.153±0.2, 23.96±0.2, 24.947±0.2, 26.273±0.2, 27.837±0.2, 28.432±0.2, 30.143±0.2, 31.072±0.2, 31.88±0.2 and 33.982±0.2.

[0060] In some embodiments, the X-ray powder diffraction pattern of the phosphate crystal form II has characteristic diffraction peaks expressed in 2θ (°) values ​​and d values ​​as shown in Table 4, and the relative intensity of each peak is shown in Table 4.

[0061] Table 4. 2θ(°), d value and relative intensity I / I0 of phosphate crystal form II

[0062] In some embodiments, the X-ray powder diffraction pattern (XRPD pattern) of the phosphate salt Form II is substantially as shown in FIG10 .

[0063] In some embodiments, the differential scanning calorimetry curve of the phosphate salt crystal form II has an endothermic peak at 207.48 ± 3 ° C, ± 2 ° C, ± 1 ° C or ± 0.5 ° C. In some embodiments, the starting temperature in the differential scanning calorimetry curve of the phosphate salt crystal form II is 207.48 ° C ± 3 ° C, 207.48 ° C ± 2 ° C, 207.48 ° C ± 1 ° C or 207.48 ° C ± 0.5 ° C, and the peak temperature is 214.31 ° C ± 3 ° C, 214.31 ° C ± 2 ° C, 214.31 ° C ± 1 ° C or 214.31 ° C ± 0.5 ° C. In some embodiments, the differential scanning calorimetry analysis curve (DSC curve) of the phosphate salt crystal form II is substantially as shown in Figure 11. In the embodiment shown in Figure 11, the melting point of the phosphate salt crystal form II of the compound of formula (I) is 207.48 ± 0.5 ° C.

[0064] In some embodiments, the thermogravimetric analysis curve (TGA curve) of the phosphate salt of the compound of formula (I), Form II, is substantially as shown in Figure 11. In some embodiments, the TGA curve of the phosphate salt of the compound of formula (I), Form II, shows almost no weight loss near 100°C, and begins to melt when heated to a melting point near 207.48°C. In some embodiments, the TGA curve of the phosphate salt of the compound of formula (I), Form II, shows that the phosphate salt is an anhydrate.

[0065] In some embodiments, the dynamic moisture absorption spectrum (DVS graph) of the phosphate salt of the compound of formula (I) Form II is substantially as shown in Figure 12. In some embodiments, the DVS graph of the phosphate salt of the compound of formula (I) Form II shows that under 80% relative humidity (RH), the moisture absorption weight gain is 0.45%.

[0066] In some embodiments, a microscopic image of Form II of the phosphate salt of the compound of formula (I) is substantially as shown in Figure 13. In some embodiments, a microscopic image of Form II of the phosphate salt of the compound of formula (I) shows that Form II of the phosphate salt is rod-shaped.

[0067] In a second aspect of the present disclosure, a method for preparing L-tartrate of the compound of formula (I) is provided.

[0068] In some embodiments, the preparation method of the L-tartrate salt of the compound of formula (I) comprises the following steps: reacting the compound of formula (I) with L-tartaric acid to form the L-tartrate salt of the compound of formula (I).

[0069] In some embodiments, in the preparation method of the L-tartrate salt, the molar ratio of L-tartaric acid to the compound of formula (I) is (0.8-1.4): 1. In some embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) is (0.9-1.2): 1.

[0070] In some embodiments, in the method for preparing the L-tartrate salt, the molar ratio of L-tartaric acid to the compound of formula (I) in the formed L-tartrate salt is (0.8-1.2): 1. In some embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) in the formed L-tartrate salt is (0.9-1.1): 1. In one embodiment, the molar ratio of L-tartaric acid to the compound of formula (I) in the formed L-tartrate salt is 1:1.

[0071] In some embodiments, a method for preparing a polymorph of an L-tartrate salt of a compound of formula (I) is also provided, wherein the polymorph is L-tartrate salt form I, comprising the following steps:

[0072] The compound of formula (I) and L-tartaric acid are subjected to a salt-forming reaction in an organic solvent to form a reaction solution;

[0073] The reaction solution is slowly cooled to obtain the L-tartrate salt form I.

[0074] In some embodiments, the organic solvent is selected from one or more of ethanol, acetonitrile, ethyl acetate, acetone, and methanol.

[0075] In some embodiments, in the preparation method of the L-tartrate salt form I, the molar ratio of L-tartaric acid to the compound of formula (I) is (0.8-1.4): 1. In other embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) is (0.9-1.2): 1.

[0076] In some embodiments, in the preparation method of the L-tartrate salt crystalline Form I, the molar ratio of L-tartaric acid to the compound of formula (I) in the obtained L-tartrate salt crystalline Form I is (0.8-1.2):1. In other embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) in the obtained L-tartrate salt crystalline Form I is (0.9-1.1):1. In one embodiment, the molar ratio of L-tartaric acid to the compound of formula (I) in the obtained L-tartrate salt crystalline Form I is 1:1.

[0077] In some embodiments, a method for preparing L-tartrate salt form II is also provided, comprising the following steps:

[0078] The compound of formula (I) and L-tartaric acid are subjected to a salt-forming reaction in an organic solvent to form a reaction solution;

[0079] The reaction solution is slowly cooled, and an anti-solvent is added to obtain the L-tartrate salt form II.

[0080] In some embodiments, the organic solvent is selected from one or more of ethanol, acetonitrile, ethyl acetate, acetone, and methanol.

[0081] In some embodiments, the anti-solvent is selected from one or more of methyl tert-butyl ether, petroleum ether, n-heptane, n-hexane, cyclohexane, isopropanol, acetone, acetonitrile, and ethyl acetate, and the anti-solvent is different from the organic solvent. In other embodiments, the anti-solvent is selected from one or more of methyl tert-butyl ether, n-heptane, isopropanol, and acetone.

[0082] In some embodiments, in the preparation method of the L-tartrate salt form II, the molar ratio of L-tartaric acid to the compound of formula (I) is (0.8-1.4): 1. In other embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) is (0.9-1.2): 1.

[0083] In some embodiments, in the preparation method of the L-tartrate salt Form II, the molar ratio of L-tartaric acid to the compound of formula (I) in the obtained L-tartrate salt Form II is (0.8-1.2): 1. In other embodiments, the molar ratio of L-tartaric acid to the compound of formula (I) in the obtained L-tartrate salt Form II is (0.9-1.1): 1. In one embodiment, the molar ratio of L-tartaric acid to the compound of formula (I) in the obtained L-tartrate salt Form II is 1:1.

[0084] In some embodiments, the temperature of the salt-forming reaction is -10°C to 90°C. For example, the temperature of the salt-forming reaction can be any range (including the end values) selected from the end values ​​of -10°C, -5°C, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C and 90°C, without limitation.

[0085] In some embodiments, the reaction solution is slowly cooled to -10°C to 60°C. In some embodiments, the reaction solution is slowly cooled to 0°C to room temperature. In some embodiments, the reaction solution is slowly cooled to room temperature.

[0086] In some embodiments, the molar concentration of the L-tartaric acid is 0.1-5 mol / L. For example, the molar concentration of the L-tartaric acid can be 0.1 mol / L, 1 mol / L, 1.5 mol / L, 2 mol / L, 2.5 mol / L, 3 mol / L, 3.5 mol / L, 4 mol / L, 4.5 mol / L, or 5 mol / L. In other embodiments, the molar concentration of the L-tartaric acid is 0.5-3 mol / L. In other embodiments, the molar concentration of the L-tartaric acid is 1-2 mol / L.

[0087] In some embodiments, the preparation method of L-tartrate salt form I comprises the following steps:

[0088] (BI-a) dissolving the compound of formula (I) in a solvent, adding an aqueous L-tartaric acid solution, and stirring to react; and

[0089] (BI-b) Cooling the reaction solution obtained in step (BI-a), precipitating a solid, performing solid-liquid separation, and collecting the solid phase to obtain L-tartrate crystalline form I of the compound of formula (I).

[0090] In some embodiments, the temperature of the reaction in step (BI-a) is 10°C to 60°C.

[0091] In some embodiments, the molar ratio of the compound of formula (I) in step (BI-a) to the L-tartaric acid contained in the L-tartaric acid aqueous solution is 1:(0.8-1.4). In other embodiments, the molar ratio of the compound of formula (I) in step (BI-a) to the L-tartaric acid contained in the L-tartaric acid aqueous solution is 1:(0.9-1.2).

[0092] In some embodiments, the molar concentration of the L-tartaric acid aqueous solution in step (BI-a) is 0.1-5 mol / L. In other embodiments, the molar concentration of the L-tartaric acid aqueous solution in step (BI-a) is 0.5-3 mol / L. In other embodiments, the molar concentration of the L-tartaric acid aqueous solution in step (BI-a) is 1-2 mol / L.

[0093] In some embodiments, the solvent in step (BI-a) is ethanol.

[0094] In some embodiments, the mixture is stirred at 30-60° C. in step (BI-a), followed by stirring at room temperature. In some embodiments, the mixture is stirred at 30-60° C. for 2-5 hours and then stirred at room temperature. In some embodiments, the mixture is stirred at 30-60° C. for 2-5 hours and then stirred at room temperature for 7-16 hours.

[0095] In some embodiments, the reaction solution is cooled to -10°C to 10°C in step (BI-b). In some embodiments, the reaction solution is cooled to -5°C to 5°C. In some embodiments, the reaction solution is cooled to about 0°C.

[0096] In some embodiments, the reaction solution is cooled for 0.25-3 hours in step (B1-b). In some embodiments, the reaction solution is cooled for 0.5-2 hours. In some embodiments, the reaction solution is cooled for 0.5-1 hour.

[0097] In some embodiments, the separation method in step (BI-b) is selected from centrifugation and filtration.

[0098] In some embodiments, after the separation in step (BI-b), the method further comprises the step of evaporating the solvent or drying.

[0099] In some embodiments, the drying step after separation in step (BI-b) is performed at 25-70°C.

[0100] In some embodiments, the preparation method of L-tartrate salt form II comprises the following steps:

[0101] (BII-a) dissolving the compound of formula (I) in a solvent, adding an aqueous L-tartaric acid solution, and stirring to react;

[0102] (BII-b) Cooling the reaction solution obtained in step (BII-a), adding an anti-solvent for crystallization, performing solid-liquid separation, and collecting the solid phase to obtain L-tartrate salt form II of the compound of formula (I).

[0103] In some embodiments, the reaction temperature in step (BII-a) is 10-60°C.

[0104] In some embodiments, the molar ratio of the compound of formula (I) to the L-tartaric acid contained in the L-tartaric acid aqueous solution in step (BII-a) is 1:(0.8-1.4). In other embodiments, the molar ratio of the compound of formula (I) to the L-tartaric acid contained in the L-tartaric acid aqueous solution is 1:(0.9-1.2).

[0105] In some embodiments, the molar concentration of the L-tartaric acid aqueous solution in step (BII-a) is 0.1-5 mol / L. In some embodiments, the molar concentration of the L-tartaric acid aqueous solution is 0.5-3 mol / L. In other embodiments, the molar concentration of the L-tartaric acid aqueous solution is 1-2 mol / L.

[0106] In some embodiments, the solvent in step (BII-a) is methanol.

[0107] In some embodiments, the mixture is stirred at 30-60° C. in step (BII-a), followed by stirring at room temperature. In some embodiments, the mixture is stirred at 30-60° C. for 2-5 h, followed by stirring at room temperature. In some embodiments, the mixture is stirred at 30-60° C. for 2-5 h, followed by stirring at room temperature for 7-16 h.

[0108] In some embodiments, the reaction solution is cooled to -10°C to 10°C in step (BII-b). In some embodiments, the reaction solution is cooled to -5°C to 5°C. In some embodiments, the reaction solution is cooled to about 0°C.

[0109] In some embodiments, the reaction solution is cooled for 0.25-3 hours in step (BII-b). In some embodiments, the reaction solution is cooled for 0.5-2 hours. In some embodiments, the reaction solution is cooled for 0.5-1 hour.

[0110] In certain embodiments, in step (BII-b), the anti-solvent is selected from one or more of methyl tert-butyl ether, normal heptane, normal hexane, cyclohexane, acetone, acetonitrile and ethyl acetate, and the solvent is different from the anti-solvent. In certain embodiments, the anti-solvent is selected from one or more of methyl tert-butyl ether, acetone, acetonitrile and ethyl acetate. In certain embodiments, the anti-solvent is methyl tert-butyl ether.

[0111] In some embodiments, the separation method in step (BII-b) is selected from centrifugation and filtration.

[0112] In some embodiments, after the separation in step (BII-b), the step of evaporating the solvent or drying is further included.

[0113] In some embodiments, the drying step after separation in step (BII-b) is performed at 25-70°C.

[0114] In some embodiments, the preparation method of L-tartrate amorphous form comprises the following steps:

[0115] (Ca) dissolving the compound of formula (I) in a solvent, adding an aqueous L-tartaric acid solution, and stirring to react;

[0116] (Cb) Cooling the reaction solution obtained in step (Ca) to precipitate a solid, performing solid-liquid separation, and collecting the solid phase to obtain an amorphous L-tartrate salt.

[0117] In some embodiments, the reaction temperature in step (Ca) is 10-60°C.

[0118] In some embodiments, the molar ratio of the compound of formula (I) in step (Ca) to the L-tartaric acid contained in the L-tartaric acid aqueous solution is 1:(0.8-1.4). In some embodiments, the molar ratio of the compound of formula (I) in step (Ca) to the L-tartaric acid contained in the L-tartaric acid aqueous solution is 1:(0.9-1.2).

[0119] In some embodiments, the molar concentration of the L-tartaric acid aqueous solution in step (Ca) is 0.1-5 mol / L. In some embodiments, the molar concentration of the L-tartaric acid aqueous solution is 0.5-3 mol / L. In some embodiments, the molar concentration of the L-tartaric acid aqueous solution is 1-2 mol / L.

[0120] In some embodiments, the solvent in step (Ca) is acetone.

[0121] In some embodiments, the mixture is stirred at 30-60° C. in step (Ca), followed by stirring at room temperature. In some embodiments, the mixture is stirred at 30-60° C. for 2-5 hours and then stirred at room temperature. In some embodiments, the mixture is stirred at 30-60° C. for 2-5 hours and then stirred at room temperature for 7-16 hours.

[0122] In some embodiments, in step (Cb), the reaction solution is cooled to -10°C to 10°C. In some embodiments, the reaction solution is cooled to -5°C to 5°C. In some embodiments, the reaction solution is cooled to about 0°C.

[0123] In some embodiments, the reaction solution is cooled for 0.25-3 hours in step (Cb). In some embodiments, the reaction solution is cooled for 0.5-2 hours. In some embodiments, the reaction solution is cooled for 0.5-1 hour.

[0124] In some embodiments, the separation method in step (Cb) is selected from centrifugation and filtration.

[0125] In some embodiments, after the separation in step (Cb), the step of evaporating the solvent or drying is further included.

[0126] In some embodiments, the drying step after separation in step (Cb) is performed at 25-70°C.

[0127] The third aspect of the present disclosure provides a method for preparing the phosphate salt of the compound of formula (I).

[0128] The preparation method of the phosphate of the compound of formula (I) comprises the following steps: subjecting the compound of formula (I) to a salt-forming reaction with phosphoric acid in the presence of an organic solvent to form the phosphate of the compound of formula (I).

[0129] In some embodiments, the phosphate is a second type phosphate of the compound of formula (I). In the step, the molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.8-1.2). For example, the molar ratio of the compound of formula (I) to phosphoric acid can be 1:0.8, 1:0.9, 1:1.0, 1:1.1, or 1:1.2.

[0130] In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.9-1.1). In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid is 1:1. In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid is 1:0.95.

[0131] In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate is 1:(0.8-1.2). In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate is 1:(0.9-1.1). In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate can be 1:0.8, 1:0.9, 1:1, 1:1.1 or 1:1.2. In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate is 1:1.

[0132] In some embodiments, the organic solvent is selected from one or more of ethanol, ethyl acetate, acetonitrile, and acetone. In some embodiments, the organic solvent is ethanol, acetone, or a mixture thereof. In some embodiments, the organic solvent is ethanol.

[0133] In some embodiments, the organic solvent is one or more of methanol, ethyl acetate, and acetone. In some embodiments, the organic solvent is one or more of methanol and ethyl acetate. In some embodiments, the organic solvent is a mixture of methanol and ethyl acetate.

[0134] In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate salt of the compound of formula (I) is 1:1.

[0135] In some embodiments, a method for preparing phosphate crystal form II is also provided, comprising the following steps:

[0136] (i) reacting the compound of formula (I) with phosphoric acid in ethanol, methanol and / or acetone to form a salt to precipitate a solid; and

[0137] (ii) collecting the solid to obtain phosphate crystal form II;

[0138] The molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.8-1.2), for example, 1:0.8, 1:0.9, 1:1.0, 1:1.1, or 1:1.2. In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.9-1.1). In one embodiment, the molar ratio of the compound of formula (I) to phosphoric acid is 1:0.95. In another embodiment, the molar ratio of the compound of formula (I) to phosphoric acid is 1:1.

[0139] In some embodiments, the salt-forming reaction is carried out in one or both of ethanol and acetone.

[0140] In some embodiments, the salt-forming reaction is carried out in ethanol. In some embodiments, the salt-forming reaction is carried out in methanol.

[0141] In some embodiments, the preparation method of phosphate crystal form II comprises the following steps:

[0142] (i-1) reacting an ethanol solution of the compound of formula (I) with an ethanol solution of phosphoric acid to form a salt to precipitate a solid; and

[0143] (ii-1) collecting the solid to obtain phosphate crystal form II;

[0144] The molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.8-1.2), for example, 1:0.8, 1:0.9, 1:1.0, 1:1.1, or 1:1.2. In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.9-1.1). In one embodiment, the molar ratio of the compound of formula (I) to phosphoric acid is 1:1.

[0145] In some embodiments, the preparation method of phosphate crystal form II comprises the following steps:

[0146] (i-1) reacting an ethanol solution of the compound of formula (I) with an ethanol solution of phosphoric acid at reflux temperature to form a salt, cooling the temperature to -10°C to 60°C to precipitate a solid; and

[0147] (ii-1) collecting the solid to obtain phosphate crystal form II;

[0148] The molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.8-1.2), for example, 1:0.8, 1:0.9, 1:1.0, 1:1.1, or 1:1.2. In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.9-1.1). In one embodiment, the molar ratio of the compound of formula (I) to phosphoric acid is 1:1.

[0149] In some embodiments, the molar ratio of the compound of formula (I) to phosphoric acid in the phosphate salt form II is 1:1.

[0150] In some embodiments, the temperature of the salt-forming reaction is -10°C to 90°C. For example, it can be any range (including the end values) consisting of the end values ​​selected from -10°C, -5°C, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C and 90°C, without specific limitation.

[0151] In some embodiments, the solids collected in step (ii) or (ii-1) can be separated by centrifugation or filtration.

[0152] In some embodiments, after collecting the solid in step (ii) or (ii-1), the step further includes evaporating the solvent or drying.

[0153] In some embodiments, the drying step is performed at 25°C to 70°C.

[0154] In some embodiments, the phosphate is a first type phosphate of the compound of formula (I). The molar ratio of the compound of formula (I) to phosphoric acid in the step is 1:(1.8-2.6), for example, 1:1.8, 1:1.9, 1:2.0, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5, or 1:2.6, etc.; preferably 1:(1.9-2.3), more preferably 1:(2.0-2.2).

[0155] In some embodiments, the molar ratio of phosphoric acid to the compound of formula (I) in the phosphate salt is (1.8-2.4):1. In some embodiments, the molar ratio of phosphoric acid to the compound of formula (I) may be 1.8:1, 1.9:1, 2.0:1, 2.1:1, 2.2:1, 2.3:1, or 2.4:1. In some embodiments, the molar ratio of phosphoric acid to the compound of formula (I) is (1.9-2.3):1. In one embodiment, the molar ratio of phosphoric acid to the compound of formula (I) is 2:1.

[0156] In some embodiments, the organic solvent is selected from one or more of ethanol, ethyl acetate, acetone, and methanol.

[0157] In some embodiments, the molar ratio of phosphoric acid to the compound of formula (I) in the phosphate salt of the compound of formula (I) is 2:1.

[0158] In one embodiment, the method for preparing the phosphate salt crystalline form I of the compound of formula (I) comprises:

[0159] (AI-a) dissolving the compound of formula (I) in an organic solvent, adding an aqueous phosphoric acid solution, and stirring to react;

[0160] (AI-b) The reaction solution obtained in step (AI-a) is cooled, an anti-solvent is added for crystallization, the solid-liquid separation is performed, and the solid phase is collected to obtain the phosphate crystal form I of the compound of formula (I).

[0161] In one embodiment, the reaction temperature in step (AI-a) is 10-60°C.

[0162] In one embodiment, the molar ratio of the compound of formula (I) to the phosphoric acid contained in the phosphoric acid aqueous solution is 1:(1.8-2.6), for example, it can be 1:1.8, 1:1.9, 1:2.0, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5 or 1:2.6, etc.; preferably it is 1:(1.9-2.3), and more preferably it is 1:(2.0-2.2).

[0163] In one embodiment, the molar concentration of the phosphoric acid aqueous solution is 0.1-5 mol / L, preferably 0.5-3 mol / L, more preferably 1-2 mol / L.

[0164] In some embodiments, in step (AI-b), the reaction solution is cooled to -10°C to 10°C, preferably -5°C to 5°C; in some embodiments, the reaction solution is cooled to about 0°C.

[0165] In some embodiments, the reaction solution is cooled for 0.5-2 h in step (AI-b); in some embodiments, the reaction solution is cooled for 0.5 h.

[0166] In one embodiment, the anti-solvent is selected from one or more of methyl tert-butyl ether, petroleum ether, n-heptane, n-hexane, cyclohexane, isopropanol, acetone, acetonitrile and ethyl acetate, and the anti-solvent is different from the solvent; preferably, the anti-solvent is selected from one or more of methyl tert-butyl ether, n-heptane, isopropanol and acetone.

[0167] In one embodiment, the separation method in step (AI-b) is selected from centrifugal separation and filtration separation.

[0168] In one embodiment, the separation in step (AI-b) further comprises the step of evaporating the solvent or drying.

[0169] In one embodiment, the drying step after separation in step (AI-b) is carried out at 25-70°C; for example, it can be 25°C, 30°C, 40°C, 50°C, 60°C or 70°C, etc., without specific limitation.

[0170] In one embodiment, the reaction temperature in step (AI-a) is 10-60°C; and / or the molar ratio of the compound of formula (I) to the phosphoric acid contained in the phosphoric acid aqueous solution is 1:(1.8-2.4), preferably 1:(1.9-2.3), more preferably 1:(2.0-2.2); and / or the molar concentration of the phosphoric acid aqueous solution is 0.1-5 mol / L, preferably 0.5-3 mol / L, more preferably 1-2 mol / L; and / or in step (AI-b), the reaction solution is cooled to -10°C-10°C, preferably -5°C-5°C, more preferably about 0°C; and / or in step (AI-b), the reaction solution is cooled to The time is 0.5-2h, more preferably 0.5h; and / or the anti-solvent is selected from one or more of methyl tert-butyl ether, petroleum ether, n-heptane, n-hexane, cyclohexane, isopropanol, acetone, acetonitrile and ethyl acetate, and the anti-solvent is different from the solvent; preferably, the anti-solvent is selected from one or more of methyl tert-butyl ether, n-heptane, isopropanol and acetone; and / or the separation method in step (AI-b) is selected from centrifugal separation and filtration separation; and / or the separation in step (AI-b) further includes a step of evaporating the solvent or drying; and / or the drying step after separation in step (AI-b) is carried out at 25-70°C.

[0171] A fourth aspect of the present disclosure provides a pharmaceutical composition comprising:

[0172] (a) L-tartrate or phosphate of the compound of formula (I) according to the first aspect of the present disclosure; and (b) a pharmaceutically acceptable carrier.

[0173] In some embodiments, the pharmaceutical composition comprises (a) the L-tartrate salt of the compound of formula (I), the first type phosphate salt of the compound of formula (I), or the second type phosphate salt of the compound of formula (I) described in the first aspect of the present disclosure; and (b) a pharmaceutically acceptable carrier.

[0174] In some embodiments, the pharmaceutical composition comprises (a) the L-tartrate salt form I of the compound of formula (I), the L-tartrate salt form II of the compound of formula (I), the L-tartrate salt amorphous form of the compound of formula (I), the phosphate salt form I of the compound of formula (I), or the phosphate salt form II of the compound of formula (I) as described in the first aspect of the present disclosure; and (b) a pharmaceutically acceptable carrier.

[0175] The fifth aspect of the present disclosure provides the use of a pharmaceutically acceptable salt of the compound of formula (I) described in the first aspect of the present disclosure, or the pharmaceutical composition described in the fourth aspect of the present disclosure in the preparation of a kinase inhibitor. Specifically, the pharmaceutically acceptable salt of the compound of formula (I) includes an L-tartrate salt of the compound of formula (I), a first type of phosphate salt of the compound of formula (I), and a second type of phosphate salt of the compound of formula (I). In some embodiments, the pharmaceutically acceptable salt of the compound of formula (I) includes an L-tartrate salt crystalline form I of the compound of formula (I), an L-tartrate salt crystalline form II of the compound of formula (I), an L-tartrate salt amorphous form of the compound of formula (I), a phosphate salt crystalline form I of the compound of formula (I), and a phosphate salt crystalline form II of the compound of formula (I). In some embodiments, the kinase inhibitor is a CDK9 inhibitor.

[0176] The sixth aspect of the present disclosure provides a pharmaceutically acceptable salt of the compound of formula (I) described in the first aspect of the present disclosure, or the pharmaceutical composition described in the fourth aspect of the present disclosure in the preparation of a drug for treating and / or preventing a disease associated with CDK9 activity or mediated by CDK9 activity. Specifically, the pharmaceutically acceptable salt of the compound of formula (I) includes an L-tartrate salt of the compound of formula (I), a first type of phosphate salt of the compound of formula (I), and a second type of phosphate salt of the compound of formula (I). In some embodiments, the pharmaceutically acceptable salt of the compound of formula (I) includes an L-tartrate salt crystalline form I of the compound of formula (I), an L-tartrate salt crystalline form II of the compound of formula (I), an L-tartrate salt amorphous form of the compound of formula (I), a phosphate salt crystalline form I of the compound of formula (I), and a phosphate salt crystalline form II of the compound of formula (I).

[0177] The present disclosure also provides a pharmaceutically acceptable salt of the compound of formula (I) according to the first aspect of the present disclosure, or a pharmaceutical composition according to the fourth aspect of the present disclosure for treating and / or preventing diseases associated with or mediated by CDK9 activity.

[0178] The present disclosure also provides a method for inhibiting CDK9 activity, comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of formula (I) described in the first aspect of the present disclosure, or the pharmaceutical composition described in the fourth aspect of the present disclosure.

[0179] The present disclosure also provides a method for treating a disease associated with or mediated by CDK9 activity, comprising administering to a subject in need thereof an effective amount of a pharmaceutically acceptable salt of a compound of formula (I) as described in the first aspect of the present disclosure, or the pharmaceutical composition as described in the fourth aspect of the present disclosure.

[0180] In some embodiments, the disease comprises a hyperproliferative disease, a virus-induced infectious disease, and a cardiovascular disease.

[0181] In some embodiments, the disease is a hyperproliferative disease. In some embodiments, the hyperproliferative disease includes angiogenesis or angiogenic proliferative disorders, mesangial cell proliferative diseases, and solid tumors. The solid tumors are, for example, cancers of the breast, respiratory tract, brain, reproductive organs, digestive tract, urinary tract, eye, liver, skin, head and neck, thyroid or parathyroid glands, and their distal metastases. In some embodiments, the disease is selected from one or more of lymphoma, sarcoma, and leukemia. In some embodiments, the disease is cancer. The cancer is, for example, pancreatic cancer, breast cancer, ovarian cancer, cervical cancer, or leukemia. In some embodiments, the disease includes solid tumors and hematologic tumors.

[0182] In the present disclosure, diseases associated with or mediated by CDK9 activity include diseases associated with or involving CDK9 activity (e.g., overactivity of CDK9), and conditions associated with these diseases. Overactivity of CDK9 refers to increased CDK9 enzymatic activity compared to normal, non-disease cells, or increased CDK9 activity that leads to unwanted cell proliferation, or reduced or insufficient programmed cell death (apoptosis), or mutations that lead to constitutive activation of CDK9.

[0183] Hyperproliferative diseases include undesirable or uncontrolled proliferative diseases involving cells, and include diseases involving reduced or insufficient programmed cell death (apoptosis). Pharmaceutically acceptable salts of compounds of formula (I) of the present disclosure or pharmaceutical compositions containing pharmaceutically acceptable salts of compounds of formula (I) can be used to prevent, inhibit, block, reduce, reduce, control, etc., and / or produce apoptosis. Treatment and or prevention of diseases associated with or mediated by CDK9 activity include administering to a subject in need thereof (including mammals, such as humans) a certain amount of pharmaceutically acceptable salts of compounds of formula (I) of the present disclosure or pharmaceutical compositions containing pharmaceutically acceptable salts of compounds of formula (I) that effectively treat or prevent the disease.

[0184] In the present disclosure, the phosphate and L-tartrate salts of the compound of formula (I) have higher solubility than the free base, and the L-tartrate salt crystal form I of the compound of formula (I), the L-tartrate salt crystal form II of the compound of formula (I), the L-tartrate salt amorphous form of the compound of formula (I), the phosphate salt crystal form I of the compound of formula (I) and the phosphate salt crystal form II of the compound of formula (I) all have good physical stability and are suitable for drug development. In particular, the phosphate salt crystal form I of the compound of formula (I) can be converted into the more stable phosphate salt crystal form II under suitable conditions, such as in ethanol. Compared with the phosphate salt crystal form I of the compound of formula (I), the phosphate salt crystal form II of the compound of formula (I) has lower hygroscopicity and better crystallinity and stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0185] In order to more clearly illustrate the embodiments of the present disclosure or the technical solutions in the related technologies, the following briefly introduces the drawings required for use in the embodiments or related technical descriptions. Obviously, the drawings described below are merely embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on the disclosed drawings without any creative work.

[0186] FIG1 is an X-ray powder diffraction (XRPD) pattern of the phosphate crystal form I of the compound of formula (I).

[0187] FIG2 is a graph of differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) of the phosphate salt form I of the compound of formula (I).

[0188] FIG3 is a dynamic water absorption curve (DVS) of the phosphate crystal form I of the compound of formula (I).

[0189] FIG4 is a microscopic image of the phosphate crystal form I of the compound of formula (I).

[0190] Figure 5 is an X-ray powder diffraction (XRPD) pattern of L-tartrate salt form I of the compound of formula (I).

[0191] FIG6 is a thermogravimetric analysis (TGA) spectrum of L-tartrate crystalline form I of the compound of formula (I).

[0192] FIG7 is an X-ray powder diffraction (XRPD) pattern of L-tartrate crystalline form II of the compound of formula (I).

[0193] FIG8 is an X-ray powder diffraction (XRPD) pattern of the amorphous L-tartrate salt of the compound of formula (I).

[0194] FIG9 is an ellipsoid diagram of the molecular stereostructure of a single crystal of the compound of formula (II).

[0195] FIG10 is an X-ray powder diffraction (XRPD) pattern of the phosphate crystal form II of the compound of formula (I).

[0196] FIG11 is a graph of differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) of Form II of the phosphate salt of the compound of formula (I).

[0197] FIG12 is a dynamic water absorption curve (DVS) of the phosphate crystal form II of the compound of formula (I).

[0198] FIG13 is a microscopic image of Form II of the phosphate salt of the compound of formula (I).

[0199] FIG14 is a comparative X-ray powder diffraction (XRPD) pattern of the conversion of the phosphate crystal form I of the compound of formula (I) into the phosphate crystal form II of the compound of formula (I) in ethanol.

[0200] Figure 15 is an XRPD diagram of the L-tartrate crystalline form I of the compound of formula (I) under high temperature (60°C) and accelerated (40°C-75% RH) conditions.

[0201] FIG16 is an XRPD diagram of the phosphate crystal form I of the compound of formula (I) under high temperature conditions.

[0202] Figure 17 is a graph showing changes in tumor volume after once-daily oral administration of Compound D and the compound of formula (I).

[0203] Figure 18 is a graph showing changes in body weight of mice after oral administration of Compound D and the compound of formula (I) once daily. DETAILED DESCRIPTION

[0204] To make the above-mentioned objects, features, and advantages of the present disclosure more clearly understood, the specific embodiments of the present disclosure are described in detail below. The following description sets forth many specific details to facilitate a full understanding of the present disclosure. However, the present disclosure can be implemented in many other ways than those described herein, and those skilled in the art may make similar modifications without violating the scope of the present disclosure. Therefore, the present disclosure is not limited to the specific embodiments disclosed below.

[0205] As used herein, the “crystal of the present disclosure,” “crystalline form of the present disclosure,” “polymorph of the present disclosure,” etc. may be used interchangeably.

[0206] In the present disclosure, the compound of formula (I) is (1S,3S)-N1-(5-((S)-1-cyclobutylethyl)pyrazolo[1,5-a]pyrimidin-7-yl)cyclopentane-1,3-diamine, and its structure is shown below:

[0207] Also included in the present disclosure are phosphate and L-tartrate salts of the compound of formula (I).

[0208] Polymorphs: Solids exist in either amorphous or crystalline forms. In the crystalline form, the molecules are arranged in a three-dimensional lattice. When a compound crystallizes from a solution or slurry, it can crystallize in different spatial arrangements (a property known as "polymorphism"), forming crystals with different crystalline forms, known as "polymorphs." Different polymorphs of a given substance can differ from one another in one or more physical properties, such as solubility and dissolution rate, true specific gravity, crystal shape, packing pattern, flowability, and / or solid-state stability.

[0209] Crystallization: Production-scale crystallization can be accomplished by manipulating the solution so that the solubility limit of the compound of interest is exceeded. This can be accomplished in a variety of ways, for example, by dissolving the compound at a relatively high temperature and then cooling the solution to below the saturation limit. Alternatively, the liquid volume can be reduced by boiling, atmospheric evaporation, vacuum drying, or other methods. The solubility of the compound of interest can be reduced by adding an antisolvent or a solvent or mixture of such solvents in which the compound has low solubility. Another alternative is to adjust the pH to reduce solubility. For a detailed description of crystallization, see Crystallization, 3rd edition, J.W. Mullens, Butterworth-Heineman Ltd., 1993, ISBN 0750611294.

[0210] The "suspension shaking" mentioned in the present disclosure refers to a method of mixing the compound of formula (I) and the corresponding acid or a solution of the corresponding acid in a suitable solvent to form a turbid solution and then shaking to obtain crystals. The suitable solvent can be water or an organic solvent.

[0211] The "suspension centrifugation" described in the present disclosure refers to a method in which the compound of formula (I) and the corresponding acid or a solution of the corresponding acid are mixed in a suitable solvent to form a turbid solution and then centrifuged to obtain crystals. The suitable solvent may be water or an organic solvent.

[0212] The "slow volatilization" mentioned in the present disclosure refers to a method of placing a solution containing the compound of formula (I) and the corresponding acid at a certain temperature to slowly volatilize the solvent to obtain crystals.

[0213] The “anti-solvent addition” or “addition of anti-solvent” mentioned in the present disclosure refers to a method of adding another suitable solvent to a solution of the compound of formula (I) and then precipitating the resulting crystals.

[0214] If it is desired that salt formation and crystallization occur simultaneously, then if the salt is less soluble in the reaction medium than the starting materials, then the addition of an appropriate acid or base can result in direct crystallization of the desired salt. Similarly, completion of the synthesis reaction in a medium in which the final desired form is less soluble than the reactants can result in direct crystallization of the final product.

[0215] Optimization of crystallization can include seeding the crystallization medium with crystals of the desired form. In addition, many crystallization methods use a combination of the above strategies. One embodiment is to dissolve the compound of interest in a solvent at high temperature, followed by the controlled addition of an appropriate volume of antisolvent to bring the system just below saturation. At this point, seeds of the desired form can be added (while maintaining the integrity of the seeds) and the system cooled to complete crystallization. As used herein, the term "about" means ±5 on the basis of a given value.

[0216] In this document, when referring to a range, if the unit is followed only by the right endpoint, it means that the units of the left and right endpoints are the same. For example, 3-5h means that the units of the left endpoint "3" and the right endpoint "5" are both h (hours).

[0217] Herein, the terms "preferred" and "better" are merely used to describe preferred implementations or examples and should not be construed as limiting the scope of protection of this disclosure. If multiple "preferred" terms appear in a technical solution, each "preferred" term is considered independent unless otherwise specified and there are no contradictions or mutual constraints.

[0218] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the any and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND" and technical solutions connected by "logical OR".

[0219] As used herein, "plurality," "multiple," "multiple times," etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "multiple" means greater than or equal to two.

[0220] In this article, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0221] As used herein, the term "polymorphs of the present disclosure" includes, but is not limited to, phosphate salt Form I of the compound of formula (I), phosphate salt Form II of the compound of formula (I), L-tartrate Form I of the compound of formula (I), and L-tartrate Form II of the compound of formula (I).

[0222] Also included in the present disclosure are phosphate salts of the compound of formula (I), particularly the monophosphate salt, and Form II of the phosphate salt of the compound of formula (I).

[0223] "Compound of formula (I)", "free base of compound of formula (I)" and "free base" are used interchangeably.

[0224] "Polymorphs of the compound of formula (I)" and "polymorphs of the free base of the compound of formula (I)" are used interchangeably.

[0225] In the present disclosure, certain crystal forms can be converted into each other, and thus the present disclosure also provides methods for converting some crystal forms into each other.

[0226] Pharmaceutical composition and its application

[0227] Generally, the pharmaceutically acceptable salts of the compounds of formula (I) disclosed herein as active ingredients can be administered together with one or more pharmaceutical carriers to form suitable dosage forms.

[0228] Specifically, the phosphate salt, L-tartrate salt, or polymorph thereof of the compound of formula (I) of the present disclosure as an active ingredient can be administered in a suitable dosage form with one or more pharmaceutically acceptable carriers.

[0229] "Pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid substance or liquid filler, diluent, encapsulating material or auxiliary formulation or any type of excipient that is compatible with the subject to which it is administered (in some embodiments, a mammal, in one embodiment, a human) and is suitable for transporting the active substance of the present disclosure to the target site without terminating its activity.

[0230] The pharmaceutical compositions of the present disclosure are formulated, dosed, and administered in a manner consistent with standard medical practice. A "therapeutically effective amount" of the active ingredient administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

[0231] The present disclosure provides pharmaceutically acceptable salts of the compound of formula (I) described in the first aspect of the present disclosure and the pharmaceutical composition described in the fourth aspect of the present disclosure, which can be used as drugs for treating and / or preventing diseases related to or mediated by CDK9 activity.

[0232] The present disclosure provides a method for inhibiting CDK9 activity, comprising administering to a subject a therapeutically effective amount of a pharmaceutically acceptable salt of the compound of formula (I) according to the first aspect of the present disclosure, or the pharmaceutical composition according to the fourth aspect of the present disclosure.

[0233] As used herein, "therapeutically effective amount" refers to an amount of a pharmaceutically acceptable salt of a compound of formula (I) of the present disclosure that will elicit a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity, or improving symptoms, alleviating symptoms, relieving or delaying disease progression, or preventing disease.

[0234] As used herein, "subject" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs and humans.

[0235] "Treatment" refers to alleviating, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition (eg, cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent, one or more symptoms of a disease or condition.

[0236] Example

[0237] The present disclosure will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present disclosure only and are not intended to limit the scope of the present disclosure. Experimental methods in the following examples, for which specific conditions are not specified, are generally performed under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight. Unless otherwise defined, the terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present disclosure.

[0238] Reagents and instruments

[0239] In the present disclosure, the structure and purity of the compounds were determined by nuclear magnetic resonance ( 1 H NMR) and / or liquid chromatography-mass spectrometry (LC-MS).

[0240] 1 H NMR: Bruker AVANCE-400 nuclear magnetic spectrometer, internal standard is tetramethylsilane (TMS).

[0241] LC-MS: Agilent 1290 HPLC System / 6130 / 6150 MS liquid chromatography-mass spectrometer (manufacturer: Agilent), column: Waters BEH / CHS, 50×2.1 mm, 1.7 μm.

[0242] HPLC analysis was performed using an Agilent 1260 Infinity HPLC, OpenLAB CDS Chemstation workstation, an XBridge C18 4.6*250 mm column, ID 5 μm column, and a DAD detector.

[0243] Elemental analysis was performed using an inductively coupled plasma optical emission spectrometer (ICP 500) with a power of 1300 W and a flow rate of 1 mL / min.

[0244] Known starting materials can be synthesized by methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0245] As used herein, room temperature in the following examples refers to about 20-30°C.

[0246] General Methods

[0247] X-ray powder diffraction (XRPD): In the present disclosure, the powder X-ray diffraction pattern of the above-mentioned crystalline or amorphous form is obtained by a known method in the art using an ARL Equinox3000 X-ray powder diffraction analyzer. The XRPD test parameters are shown in Table 5 below.

[0248] Table 5

[0249] In the powder X-ray diffraction pattern, the position of each peak is determined by the 2θ (°) value. It is understood that different instruments and / or conditions may result in slightly different data, and the position and relative intensity of each peak may vary.

[0250] The intensity division of peak only reflects the approximate size of peak in each position. In the present disclosure, each crystal form is used as the base peak with the highest diffraction peak of its peak height, and its relative intensity is defined as 100%, as I0 (the peak with 2θ (°) value of phosphate crystal form I is 18.803 as the base peak, the peak with 2θ (°) value of L-tartrate crystal form I is 20.27 as the base peak, the peak with 2θ (°) value of L-tartrate crystal form II is 21.353 as the base peak, and the peak with 2θ (°) value of phosphate crystal form II is 18.230 as the base peak), and the ratio of its peak height to the base peak height of each other peak is used as its relative intensity I / I0, and the division definition of each peak relative intensity is shown in Table 6 below.

[0251] Table 6

[0252] Single Crystal X-ray Diffraction (SXRD): In the present disclosure, single crystal X-ray diffraction patterns of the compound of Formula (II) were obtained using a D8 Venture diffractometer using methods known in the art. The SXRD test parameters are shown in Table 7. After collecting relevant data, the crystal structure was further analyzed using a direct method (SHELXT2014) to confirm the absolute configuration.

[0253] Table 7

[0254] The phosphate salt of the compound of formula (I) disclosed herein and its crystal form were determined by elemental analysis to determine the acid-base molar ratio, and the L-tartrate salt of the compound of formula (I) and its crystal form and its amorphous form were determined by HPLC / IC or 1 The acid-base molar ratio was determined by H NMR.

[0255] High Performance Liquid Chromatography: In this disclosure, high performance liquid chromatography (HPLC) was acquired on an Agilent 1260 HPLC.

[0256] Differential Scanning Calorimetry (DSC): In the present disclosure, the differential scanning calorimetry spectrum of the above-mentioned crystal form is obtained by a method known in the art using a DSC25A differential scanning calorimeter. The DSC test parameters are shown in Table 8 below.

[0257] Table 8

[0258] Thermogravimetric analysis (TGA): In the present disclosure, the thermogravimetric analysis spectrum of the above-mentioned crystal form is obtained by a known method in the art using a TGA550 thermogravimetric analyzer. The TGA test parameters are shown in Table 9 below.

[0259] Table 9

[0260] Dynamic moisture sorption (DVS) curves were collected on an SMS (Surface Measurement Systems) DVS Intrinsic instrument. Relative humidity at 25°C was calibrated using the deliquescent points of LiCl, Mg(NO₃)₂, and KCl. Instrument test conditions are shown in Table 10 below.

[0261] Table 10

[0262] The "Guidelines for Hygroscopicity Tests of Drugs" No. 9103 of the "Chinese Pharmacopoeia" (2020) stipulates the description of hygroscopic characteristics and the definition of hygroscopic weight gain: (1) Deliquescent: absorbing sufficient water to form a liquid; (2) Extremely hygroscopic: hygroscopic weight gain is not less than 15%; (3) Hygroscopic: hygroscopic weight gain is less than 15% but not less than 2%; (4) Slightly hygroscopic: hygroscopic weight gain is less than 2% but not less than 0.2%; (5) No or almost no hygroscopic: hygroscopic weight gain is less than 0.2%.

[0263] It is understood that other types of instruments having the same function as the above-mentioned instruments or using test conditions different from those used in the present disclosure may result in different values. Therefore, the quoted values ​​should not be regarded as absolute values.

[0264] Due to instrument errors or differences in operators, those skilled in the art will understand that the above parameters used to characterize the physical properties of crystals may have slight differences. Therefore, the above parameters are only used to assist in characterizing the polymorphs provided by the present disclosure and cannot be regarded as limitations on the polymorphs provided by the present disclosure.

[0265] Unless otherwise specified, "phosphoric acid solution" and "L-tartaric acid solution" used in this disclosure refer to aqueous phosphoric acid solutions and aqueous L-tartaric acid solutions. "Phosphate" and "L-tartrate" used in this disclosure refer to the phosphate and L-tartrate salts of the compound of formula (I).

[0266] The solvent used in the present disclosure is an analytically pure solvent, such as ethanol, which is analytically pure ethanol (water content ≤ 0.3%).

[0267] Compound D in this application and compound E It can be prepared by referring to existing published patent documents.

[0268] As used herein, MTBE refers to methyl tert-butyl ether; DMSO refers to dimethyl sulfoxide; THF refers to tetrahydrofuran; EA refers to ethyl acetate; PE refers to petroleum ether; DCM refers to dichloromethane; MeOH refers to methanol; CDI refers to N,N-carbonyldiimidazole; MOPS refers to 3-(N-morpholino)propanesulfonic acid; Tween-20 refers to Tween 20; DTT refers to dithiothreitol; EDTA refers to ethylenediaminetetraacetic acid; CDK1 refers to cyclin-dependent kinase 1; CDK2 refers to cyclin-dependent kinase 2; CDK9 refers to cyclin-dependent kinase 9; K2EDTA refers to dipotassium ethylenediaminetetraacetate; IPA refers to isopropanol; and NH3 refers to ammonia.

[0269] Preparation Example 1. Preparation of Compound 6

[0270] Step 1: Dissolve sodium hydride (18 g, 450.00 mmol, 60% purity) in THF (300 mL), cool to 0°C, and slowly add triethyl 2-phosphonopropyl ester (100 g, 419.78 mmol) dropwise. The reaction is stirred at 0-5°C for 1 hour. Then, cyclobutanone (25 g, 356.69 mmol) dissolved in THF (50 mL) is slowly added dropwise. After the addition is complete, the mixture is allowed to react at 25°C for 15 hours. The reaction is quenched with saturated sodium chloride solution (600 mL) and extracted with ethyl acetate (600 mL x 2). The combined organic phases are washed with saturated sodium chloride solution (500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by CombiFlash separation (120 g x 2, 0-20% EA / PE) to afford compound 2 (44 g, colorless oil) in an 80.00% yield. 1 H NMR (400MHz, DMSO-d6) δ4.05(q,J=7.2Hz,2H),3.01-2.93(m,2H),2.78-2.71(m,2H),2.00-1.89(m,2H),1.61-1.58(m,3H),1.18(t,J=7.2Hz,3H).

[0271] Step 2: Dissolve compound 2 (44 g, 285.33 mmol) in methanol (400 mL) and add wet palladium on carbon (4.4 g, 10% purity). Cover with a hydrogen balloon and replace the hydrogen atmosphere three times. Stir at 27°C under a hydrogen atmosphere (15 psi) for 6-8 hours. TLC shows the disappearance of the starting material spot. Filter off the Pd / C and remove the solvent under reduced pressure to obtain crude compound 3 (44 g, colorless oil, boiling point approximately 182°C) in a yield of 98.71%. Continue to the next step without purification.

[0272] Step 3: Compound 3 (44 g, 281.65 mmol) was dissolved in methanol (400 mL), and sodium hydroxide (45.06 g, 1.13 mol) and water (200 mL) were added, followed by stirring at room temperature for 16 hours. LCMS indicated the reaction was complete. The methanol was removed by concentration under reduced pressure, and the mixture was then extracted twice with dichloromethane. The aqueous phase was adjusted to pH 3 with dilute hydrochloric acid (6 M), and extracted with DCM (300 mL x 3). The organic phases were combined, dried, and spin-dried to afford compound 4 (35 g, colorless oil, boiling point approximately 220°C). Yield: 96.96%, which was carried on to the next step without purification. MS m / z (ESI): 127.1 [MH] - . 1H NMR (400MHz, DMSO-d6) δ11.91 (s, 1H), 2.36-2.20 (m, 2H), 1.99-1.90 (m, 2H), 1.82-1.58 (m, 4H), 0.93 (d, J = 6.8Hz, 3H).

[0273] Step 4: In a reaction flask, compound 4 (36 g, 280.88 mmol) was dissolved in THF (300 mL), and CDI (68.32 g, 421.32 mmol) was added. The mixture was allowed to react at room temperature for 16 hours (Solution A). In a separate reaction flask, potassium monomethyl malonate (143.42 g, 842.64 mmol) was added to anhydrous magnesium chloride (66.86 g, 702.20 mmol) and THF (900 mL). The mixture was heated at 50°C under argon for 16 hours (Solution B). Solution A was then added dropwise to Solution B at room temperature (approximately 10 minutes), and the mixture was stirred at 30°C for 16 hours. LCMS confirmed the reaction was complete and the product was produced. 800 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (800 mL x 3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and then evaporated to dryness under reduced pressure. The residue was separated and purified by CombiFlash (120 g × 2, 0-15% EA / PE) to give the product, compound 5 (42 g, light yellow oil, bp ~ 240°C), yield: 75.42%. MS m / z (ESI): 199.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ4.13-4.00(m,2H),3.55(d,J=0.8Hz,2H),2.63-2.54(m,1H),2.42-2.28(m,1H) ,1.97-1.85(m,2H),1.83-1.74(m,1H),1.73-1.61(m,3H),1.16(t,J=7.2Hz,3H),0.91(d,J=6.8Hz,3H).

[0274] Step 5: Dissolve compound 5 (42 g, 211.85 mmol) and 3-aminopyrazole (19.36 g, 233.03 mmol) in glacial acetic acid (300 mL) and heat to 120°C for 16 h. LCMS indicated the reaction was complete. Concentrate under reduced pressure to remove the acetic acid, and slurry the mixture with ethyl acetate (800 mL x 4). The solid precipitated, was filtered, and dried to afford compound 6 (41 g, pale yellow solid) in an 89.08% yield. The product was carried on to the next step without purification. MS m / z (ESI): 218.1 [M+H] + .

[0275] Preparation Example 2. Preparation of the compound of formula (I)

[0276] Step 1: Dissolve compound 6 (473.6 mg, 2.18 mmol) in phosphorus oxychloride (6 mL) and heat to 120°C with stirring for 3 hours. Cool to room temperature, pour into ice water (60 g), and extract with dichloromethane (80 mL). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue is separated by silica gel column chromatography (20 g, 0%-40% EA / DCM) to provide compound 7. MS m / z (ESI): 236.1 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.25(d,J=2.4Hz,1H),7.35(s,1H),6.76(d,J=2.4Hz,1H),2.893-2 .84(m,1H),2.61-2.51(m,1H),2.12-2.01(m,1H),1.80-1.59(m,5H),1.14(d,J=6.8Hz,3H).

[0277] Step 2: Compound 7 (142.7 mg, 605.5 μmol) and tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (121.27 mg, 605.52 μmol) were dissolved in acetonitrile (20 mL), and potassium carbonate (251.0 mg, 1.81 mmol) was added. The reaction was stirred at 90°C for 16 hours. Ethyl acetate (80 mL) was added to dilute the mixture, and the mixture was washed with saturated sodium chloride solution (80 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield compound 8. MS m / z (ESI): 400.3 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.00(d,J=2.0Hz,1H),7.58(d,J=7.6Hz,1H),6.97(d,J= 7.6Hz,1H),6.30(d,J=2.0Hz,1H),6.00(s,1H),4.19(q,J=7.2Hz,1H),4.02–3.9 3(m,1H),2.77-2.66(m,1H),2.61-2.53(m,1H),2.21-2.01(m,3H),1.97-1.88(m ,2H),1.80-1.63(m,6H),1.56-1.42(m,1H),1.40(s,9H),1.13(d,J=6.8Hz,3H).

[0278] Step 3: Compound 8 (174.7 mg, 437.38 μmol) was dissolved in 1,4-dioxane (3 mL), and hydrochloric acid solution (3.0 mL, 4 M) was added. The mixture was stirred at room temperature for 3 hours. The solvent was evaporated under reduced pressure, and water (60 mL) was added. The mixture was extracted with ethyl acetate (50 mL). The aqueous phase was adjusted to pH 9-10 with saturated sodium carbonate solution and extracted with ethyl acetate (60 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 9 was isolated by preparative HPLC chromatography. MS m / z (ESI): 300.2 [M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.97(s,1H), 7.54(s,1H),6.28(s,1H),5.99(s,1H),4.13-4.22(m,1H),3.90-3.99(m,1H),2.75-2.6.3(m,1H),2.51-2.5 7(m,1H),2.20-1.96(m,3H),1.85-1.94(m,2H),1.60-1.78(m,6H),1.40-1.49(m,1H),1.11(d,J=6.8Hz,3H).

[0279] Step 4: Compound 9 (94.46 mg, 315.48 μmol) was subjected to chiral separation (IC-3 4.6*100 mm 3 μm column; cosolvent: IPA [1% NH3 (7 M in MeOH)]; injection volume: 5.00 μL; wavelength: 220.0 nm; run time: 6.0 min; flow rate: 3.0 mL / min; pressure: 2000 psi; column temperature: 40°C) to afford compound (I) (9.70 mg, retention time 2.471 min) in 9.89% yield and 96.34% purity. MS m / z (ESI): 300.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ7.96(d,J=2.0Hz,1H),7.41(d,J=7.6Hz,1H),6.27(d ,J=2.0Hz,1H),5.96(s,1H),4.21(q,J=7.2Hz,1H),3.42(q,J=6.0Hz,1H),2. 65-2.74(m,1H),2.58-2.50(m,1H),2.16-2.24(m,1H),2.03-2.09(m,1H),1. 83-1.95(m,2H),1.80-1.61(m,7H),1.35-1.25(m,1H),1.11(d,J=6.8Hz,3H).

[0280] The compound of formula (II) is prepared from the compound of formula (I), so that the absolute configuration of the compound of formula (I) can be determined by the absolute configuration of the compound of formula (II).

[0281] The compound of formula (I) (200 mg, 667.97 μmol) was dissolved in dichloromethane (10 mL), and 4-chlorobenzoyl chloride (175.4 mg, 1.00 mmol) was added, followed by N,N-diisopropylethylamine (0.6 mL, 3.34 mmol). The reaction was stirred at room temperature for 2 hours. LCMS analysis showed that the reaction was complete. Water (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL). The organic phase was washed with saturated sodium chloride solution (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography to obtain the compound of formula (II) (160 mg, yield: 54.69%). LCMS (ESI) m / z: 438.2 [M+H] + .

[0282] A single crystal of the compound of formula (II) was further prepared, and the molecular structure ellipsoid diagram of the compound of formula (II) is shown in Figure 9. The Flack constant is 0.07(2), and C8, C10, and C19 are in the S configuration, that is, the absolute configuration of the three chiral centers of the compound of formula (II) is all S configuration, and its structure is shown in the structure of the compound of formula (II). Therefore, it can be determined that the absolute configuration of the three chiral centers of the compound of formula (I) is all S configuration, and its structure is shown in the structure of the compound of formula (I).

[0283] Example 1. Preparation of Phosphate Crystalline Form I of the Compound of Formula (I)

[0284] A 20 mL sample vial was charged with free base (500 mg) and methanol (1 mL), sonicated for dissolution, and then 1 mol / L phosphoric acid solution was added at a molar ratio of 2.5:1. The reaction was allowed to proceed at 50°C for 1 hour, followed by stirring at 40°C for 1 hour, and then at 30°C for another hour. The heat was then turned off and the mixture was stirred overnight. After the reaction, the temperature was slowly lowered to 0°C, acetone was added to precipitate a solid, which was collected by centrifugation and the solvent evaporated. The resulting solid was the phosphate salt Form I of the compound of formula (I). Elemental analysis of the resulting solid revealed a phosphorus content of 12.5%, indicating a molar ratio of the compound of formula (I) to phosphoric acid of 1:2 (the theoretical phosphorus content is 12.5%). The XRPD pattern of the resulting solid is substantially as shown in Figure 1 , and the XRPD pattern of the resulting solid exhibits peaks at the 2θ (°) values ​​shown in Table 1 above, with the relative intensities of each peak as shown in Table 1 above. The DSC and TGA patterns are shown in Figure 2 , the DVS pattern is shown in Figure 3 , and the microscopic image is shown in Figure 4 .

[0285] As shown in Figure 2, the DSC graph shows a melting absorption peak at 188.01°C, indicating good crystallinity, indicating that the melting point of the phosphate form I of the compound of formula (I) is approximately 188.01°C; in the TGA graph, the sample loses 1.139% of its weight when heated to around 190°C, which may be due to solvent volatilization.

[0286] As shown in FIG3 , in the DVS graph, under 80% RH conditions, the weight gain due to moisture absorption is 9%, indicating that the phosphate crystal form I of the compound of formula (I) is hygroscopic.

[0287] As shown in FIG4 , the polarizing microscope image shows that the phosphate crystal form I of the compound of formula (I) is in the shape of rods and blocks.

[0288] Example 2. Preparation of L-tartrate Form I of the compound of formula (I)

[0289] To a 20mL sample vial, add the free base (500mg) and ethanol (1mL) and sonicate to dissolve. Then, add a 1mol / L L-tartaric acid solution at a molar ratio of 1.2:1. The reaction was allowed to proceed at 50°C for 1h, followed by stirring at 40°C for 1h, and then at 30°C for another 1h. Finally, turn off the heat and stir at room temperature overnight. After the reaction, slowly cool to 0°C, collect the solid by centrifugation, and evaporate the solvent. The resulting solid is the L-tartrate salt, Form I, of the compound of formula (I). HPLC / IC results indicate that the molar ratio of the compound of formula (I) to L-tartaric acid in the resulting solid is 1:1. The XRPD pattern is substantially as shown in Figure 5 , and the XRPD pattern exhibits peaks at the 2θ (°) values ​​shown in Table 2 above, with the relative intensities of each peak as shown in Table 2 above. The TGA pattern is shown in Figure 6 .

[0290] As shown in Figure 6, the TGA graph shows that the weight loss is 1.057% when heated to around 110°C, which may be due to the volatilization of water. The weight loss continues to be 3.482% when heated to around 178°C, which is presumably due to the volatilization of the organic solvent. Further heating causes melting and decomposition.

[0291] Example 3. Preparation of L-tartrate Form II of the compound of formula (I)

[0292] In a sample bottle, add free base (about 100 mg) and methanol (0.2 mL to 0.3 mL), sonicate to dissolve, then add 1 mol / L L-tartaric acid solution according to the ratio of acid-base molar ratio = 1.2:1, react at 50 ° C for 1 hour, continue stirring at 40 ° C for 1 hour, continue stirring at 30 ° C for another hour, turn off the heat after cooling to room temperature, and stir overnight. After the reaction is completed, slowly cool to 0 ° C, add MTBE to crystallize, and evaporate the solvent to obtain a solid. The obtained solid is subjected to HPLC / IC analysis to determine that the molar ratio of the compound of formula (I) to L-tartaric acid in the obtained solid is 1:1; the XRPD pattern of the obtained solid is shown in Figure 7, and the XRPD pattern of the obtained solid has peaks at the 2θ (°) values ​​shown in Table 3 above, and the relative intensities of each peak are shown in Table 3 above. In this disclosure, it is defined as L-tartrate salt form II of the compound of formula (I).

[0293] Example 4. Preparation of amorphous L-tartrate of compound (I)

[0294] To a sample vial, add the free base (approximately 100 mg) and acetone (0.2 mL to 0.3 mL) and sonicate to dissolve. Then, add a 1 mol / L L-tartaric acid solution at a molar ratio of 1.2:1. React at 50°C for 1 hour, continue stirring at 40°C for 1 hour, and continue stirring at 30°C for another hour. Cool to room temperature, turn off the heat, and stir overnight. After the reaction, slowly cool to 0°C to precipitate a solid. The solid was collected by centrifugation and the solvent was evaporated. The resulting solid was subjected to HPLC / IC to determine a 1:1 molar ratio of the compound of formula (I) to L-tartaric acid. The XRPD pattern of the resulting solid is shown in Figure 8, and is defined as the amorphous L-tartrate salt in this application.

[0295] Example 5. Preparation of Phosphate Form II of the Compound of Formula (I)

[0296] The free base (100 mg) was dissolved in ethanol (15 mL) in a 50 mL sample vial. An ethanolic solution of phosphoric acid (5 mL) was then added dropwise at a molar ratio of 0.95:1 between the acid and base. The mixture was stirred at room temperature for 4 hours, filtered, rinsed with ethanol, and dried by rotary evaporation to yield a white solid. Elemental analysis of the resulting solid revealed a phosphorus content of 7.8%, indicating a molar ratio of 1:1 between the compound of formula (I) and phosphoric acid (theoretical phosphorus content is 7.79%). The resulting solid was subjected to XRPD analysis, and its XRPD pattern is shown in Figure 10. The solid is defined as phosphate crystal form II in this application.

[0297] The XRPD pattern of the obtained solid has peaks at the 2θ (°) values ​​shown in Table 4 above, and the relative intensities of the peaks are shown in Table 4 above; its DSC and TGA curves are shown in Figure 11, its DVS graph is shown in Figure 12, and its microscope image is shown in Figure 13.

[0298] As shown in Figure 11, in the DSC curve, the sample has a melting absorption peak at 207.48°C, indicating good crystallinity, indicating that the melting point of the phosphate crystal form II of the compound of formula (I) is approximately 207.48°C. In the TGA curve, the sample shows almost no weight loss when heated to around 100°C, and begins to melt when heated to around 207°C. Therefore, the phosphate crystal form II of the compound of formula (I) is an anhydrate.

[0299] As shown in FIG12 , in the DVS graph, under 80% RH conditions, the weight gain due to moisture absorption was 0.45%, indicating that the phosphate crystal form II of the compound of formula (I) is slightly hygroscopic.

[0300] As shown in FIG13 , polarizing microscopy shows that the phosphate crystal form II of the compound of formula (I) is rod-shaped.

[0301] Example 6. Preparation of Phosphate Crystalline Form II of the Compound of Formula (I)

[0302] The free base (100 mg) was dissolved in acetone (15 mL) in a 50 mL sample vial. An ethanolic solution of phosphoric acid (5 mL) was then added dropwise at a molar ratio of 0.98:1. The mixture was stirred at room temperature for 4 hours, filtered, rinsed with ethanol, and dried by rotary evaporation to yield a white solid. Elemental analysis of the resulting solid revealed a phosphorus content of 7.8%, indicating a 1:1 molar ratio of the compound of formula (I) to phosphoric acid (theoretical phosphorus content is 7.79%). The resulting solid was subjected to XRPD analysis, and its XRPD pattern is generally shown in Figure 10.

[0303] Example 7. Preparation of Phosphate Crystalline Form II of the Compound of Formula (I)

[0304] The free base (100 mg) was dissolved in ethanol (10 mL) in a 50 mL sample vial. An ethanolic solution of phosphoric acid (10 mL) was then added dropwise at a molar ratio of 1.05:1 between the acid and base. The mixture was stirred at room temperature for 4 hours, filtered, rinsed with ethanol, and dried by rotary evaporation to yield a white solid. Elemental analysis of the resulting solid revealed a phosphorus content of 7.8%, indicating a 1:1 molar ratio of the compound of formula (I) to phosphoric acid (theoretical phosphorus content is 7.79%). The resulting solid was subjected to XRPD analysis, and its XRPD pattern is generally shown in Figure 10.

[0305] Example 8. Preparation of Phosphate Crystalline Form II of the Compound of Formula (I)

[0306] 0.5 g of the free base was dissolved in methanol (15 ml). Commercially available phosphoric acid was added in a 1:1 molar ratio of acid to base. After stirring for 2 hours, the methanol was removed by rotary evaporation to obtain a crude phosphate. Ethanol (2.5 ml) was added and the mixture was concentrated. Ethanol (2.5 ml) was added and the mixture was concentrated again. Ethanol (10 ml) was added and the mixture was heated to 60° C. After stirring for 3 hours, the mixture was cooled to room temperature and stirred overnight. The mixture was filtered, rinsed with ethanol, and dried by rotary evaporation to obtain the phosphate salt Form II of the compound of formula (I) in a yield of 75.8%. Elemental analysis of the resulting solid revealed a phosphorus content of 7.7%, indicating a 1:1 molar ratio of the compound of formula (I) to phosphoric acid in the resulting solid (theoretical phosphorus content is 7.79%). The resulting solid was subjected to XRPD analysis, and its XRPD pattern is generally shown in Figure 10.

[0307] Comparative Example 1. Salt-forming reaction of the compound of formula (I)

[0308] An appropriate amount of the compound of formula (I) (100 mg) was weighed using the weight loss method and placed in a transparent sample vial. The corresponding solvent (0.2 mL to 0.3 mL) was added and dissolved by sonication. A 1 mol / L acid solution (aqueous solution) was added at a molar ratio of 1.2:1 between the acid and base. The reaction was incubated at 50°C for 1 hour, followed by stirring at 40°C for 1 hour, and then at 30°C for another hour. After cooling to room temperature, the heat was turned off and stirred overnight. After the reaction was completed, the temperature was slowly lowered to allow solid precipitation. If the solution remained clear, crystallization was induced by adding an antisolvent. The reaction results were analyzed by XRPD. The results showed that acids such as hydrochloric acid, sulfuric acid, and oxalic acid failed to form salts of the compound of formula (I) after cooling or adding an antisolvent. The types of acids and solvents are shown in Table 11 below.

[0309] Table 11

[0310] Test Example 1. Solubility Test

[0311] The solubility of the phosphate and L-tartrate salts of the compound of formula (I) prepared in the above examples in water was tested at room temperature. The solubility of the phosphate salt was no greater than 26.6 mg / mL, and the solubility of the L-tartrate salt was no greater than 17.3 mg / mL. The free base was slightly soluble in water. The salts of the compound of formula (I) significantly increase its solubility in water, with the phosphate salt having a higher solubility than the L-tartrate salt.

[0312] In addition, the crude solubility (unit: mg / mL) of the phosphate salt Form I and L-tartrate salt Form I of the compound of formula (I) in a pH 4.5 buffer (acetic acid-sodium acetate system) and a pH 6.8 buffer (sodium dihydrogen phosphate-sodium hydroxide system) at room temperature was tested, and the results are shown in Table 12. As can be seen from the results in Table 12, the phosphate salt Form I and L-tartrate salt Form I of the compound of formula (I) have high solubility in both pH 4.5 and pH 6.8 buffers.

[0313] Table 12

[0314] An appropriate amount of phosphate crystal form I or phosphate crystal form II was weighed and added to 0.1 mol / L hydrochloric acid solution (5 mL) and pH 6.8 phosphate buffer (5 mL), respectively. After 24 h (placed in a 37°C oven), samples were taken to determine the solubility. The results are shown in Table 13.

[0315] Table 13

[0316] Test Example 2. Stability Test

[0317] (1) Samples (approximately 100 mg) of the phosphate crystal form I of the compound of formula (I), the phosphate crystal form II of the compound of formula (I), the L-tartrate crystal form I of the compound of formula (I), and the L-tartrate crystal form II of the compound of formula (I) were weighed respectively and placed under 60°C (high temperature) and 40°C-75% RH (accelerated) conditions. At the same time, another group of samples was sealed and stored at 5°C as a control. The crystal form changes were detected after 7 days, 20 days, 30 days, or 60 days.

[0318] The test results show that the crystal forms of the L-tartrate salt form I of the compound of formula (I), the L-tartrate salt form II of the compound of formula (I), the phosphate salt form I of the compound of formula (I), and the phosphate salt form II of the compound of formula (I) all showed no significant changes under the above conditions, and had high stability. The XRPD pattern of the crystal form of the L-tartrate salt form I of the compound of formula (I) was stored under high temperature conditions for 7 days and 2 months (60 days), and the XRPD pattern of its crystal form showed no significant changes; the superposition of the XRPD pattern of the crystal form of the L-tartrate salt form I of the compound of formula (I) was stored under accelerated conditions for 7 days and 2 months (60 days) as shown in Figure 15, showing no significant changes; this indicates that the L-tartrate salt form I of the compound of formula (I) has high stability. The superposition of the XRPD pattern of the crystal form of the phosphate salt form I of the compound of formula (I) was stored under high temperature conditions for 7 days and 30 days as shown in Figure 16, showing no significant changes, indicating that the phosphate salt form I of the compound of formula (I) has high stability.

[0319] (2) About 40 mg of phosphate crystal form I (prepared according to Example 1 of the present application) was weighed into a glass vial, ethanol (1 mL) was added, the vial was sealed, and the mixture was shaken at 50°C and 150 rpm for seven days. The solid was then collected by centrifugation and the solvent was evaporated. The resulting solid was subjected to XRPD analysis, and its XRPD pattern is shown in Figure 10. The resulting solid is phosphate crystal form II. The test results in Figure 14 show that phosphate crystal form I can be converted into the more thermodynamically stable phosphate crystal form II.

[0320] (3) 25 mg of each of phosphate crystal form I and phosphate crystal form II were taken, 5 mg of water was added to each, and then placed in an 80°C oven for one week. Samples were taken and the relevant substances were determined. The results are shown in Table 14, indicating that phosphate crystal form I and phosphate crystal form II have high chemical stability.

[0321] Table 14

[0322] Test Example 3. Hygroscopicity test

[0323] 3.1 L-Tartrate Hygroscopicity Test

[0324] The hygroscopicity test was carried out in accordance with the operation of 9103 (Guidelines for Drug Hygroscopicity Test) in the "Chinese Pharmacopoeia (2020)", and it was found that the L-tartrate salt of the compound of formula (I) was slightly hygroscopic.

[0325] 3.2 Hygroscopicity test of phosphate crystals

[0326] DVS measurements were performed on Form I and Form II of the phosphate salt of the compound of formula (I). The DVS graph of Form I of the phosphate salt of the compound of formula (I) is shown in Figure 3 . The DVS graph shows that under conditions of 80% RH, the weight gain due to moisture absorption is 9%, indicating that Form I of the phosphate salt of the compound of formula (I) is hygroscopic. The DVS graph of Form II of the phosphate salt shows that under conditions of 80% RH, the weight gain due to moisture absorption is only 0.45%, indicating that Form II of the phosphate salt is slightly hygroscopic.

[0327] Test Example 4. Activity inhibition test of CDK family kinases

[0328] In the following LANCE Ultra test method, kinase reagents were purchased from Carna Bioscience, reaction substrates and detection reagents were purchased from PerkinElmer, and the remaining reagents were purchased from Thermo Scientific.

[0329] The LANCE Ultra method was used to determine the inhibitory effect of the test substances on the kinase activities of CDK1 / CycB (Carna bioscience, #04-102), CDK2 / CycA (Carna bioscience, #04-103), and CDK9 / CycT (Carna bioscience, #04-110).

[0330] The kinase activity test used a 10 μL system containing the following components: CDK kinase diluent, a substrate diluent containing a mixture of Ulight-Myelic basic protein (PerkinElmer, #TRF-0109, hereinafter referred to as U-MBP) and ATP (Thermo Scientific, #PV3227), and the compound of formula (I) disclosed herein (i.e., the test substance). Each kinase was tested in three groups: a background group (Blank), a non-inhibitory group (PC), and a compound test group (Test). The components included in each test group are shown in Table 15 below.

[0331] Table 15

[0332] The working concentrations of the components of the Test group in different kinase reactions are shown in Table 16:

[0333] Compounds: Dissolve the test compound at 10 mM at room temperature and serially dilute it in DMSO. Then dilute it to a 4x working solution in deionized water with 2% DMSO. The highest compound concentration used in the CDK1 and CDK2 assays is 10 μM, and CDK9 is 1 μM.

[0334] 1.33x reaction buffer: Components: 26.7 mM MOPS, 6.67 mM MgCl2, and 0.0133% Tween-20. Store in a 4°C refrigerator away from light. Add freshly prepared DTT to a final concentration of 5.33 mM before use.

[0335] Table 16

[0336] The working concentration of DMSO in the reaction was 0.5%.

[0337] After mixing the above components, place them on a shaker and incubate them in the dark at room temperature for 1 hour. Then, 10 μL of detection solution was added to all test groups (including Blank, PC and Test groups).

[0338] 10 μL of detection solution contains the following components: 16 mM EDTA (Thermo scientific, #15575), 1 nM phosphorylated U-MBP protein antibody (PerkinElmer, #TRF-0201) and 1x detection buffer (PerkinElmer, #CR97-100).

[0339] After adding the test solution, place on a shaker and continue incubating at room temperature in the dark for 1 hour. After incubation, use a PerkinElmer VictorX5 fluorescence microplate reader to read the signal, with an excitation wavelength of 320 nm and emission wavelengths of 615 nm and 665 nm. The inhibition rate is calculated as follows:

[0340] 1. Calculate the 665nm / 615nm value (hereinafter referred to as the Ratio value) for all groups. The inhibition rate is calculated based on the Ratio value of each group.

[0341] 2. Inhibition rate = (PC Ratio -Test Ratio ) / (PC Ratio -Blank Ratio )*100%;

[0342] 3. XLFIT5.0 software (IDBS, UK) was used for fitting, with the logarithm of the compound concentration as the X-axis and the inhibition rate as the Y-axis, and the half-maximal inhibitory concentration IC of the compound was calculated using the four-parameter model. 50 , the results are shown in Table 17.

[0343] Table 17

[0344] From the results in Table 17, it can be seen that the compound of formula (I) disclosed in the present invention has high inhibitory activity against CDK9 and high CDK9 inhibition selectivity.

[0345] Test Example 5: In vivo pharmacokinetic study in mice

[0346] The LC / MS / MS method was used to determine the drug concentrations in the plasma of mice at different times after intravenous injection and oral administration of Compound D, Compound E and the compound of formula (I), respectively, to study the pharmacokinetic behavior of Compound D, Compound E and the compound of formula (I) in mice and evaluate their pharmacokinetic characteristics.

[0347] Experimental plan:

[0348] Experimental animals: Healthy adult male ICR mice (weight 30-40 g, 12 mice; mice in the intravenous injection group had free access to water and food; mice in the oral administration group were fasted overnight and had free access to water and food 4 h after administration) were provided by Beijing Vital River Laboratory Animal Co.LTD.

[0349] Administration and dosage: ICR mice were administered via tail vein (2 mg / kg, 5% DMSO, pH 4.5 20% Captisol) and oral administration (10 mg / kg, 5% DMSO, pH 4.5 20% Captisol).

[0350] Blood Sampling: Before dosing, select animals that meet the experimental requirements, weigh them, and label them. Before blood sampling, restrain the mice. Each dosed mouse receives blood at the designated time points (intravenous administration: 0.083, 0.25, 0.5, 1, 2, 4, 6, 7.5, and 24 hours after dosing, for a total of nine time points; oral administration: 0.083, 0.25, 0.5, 1, 2, 4, 6, 7.5, and 24 hours after dosing, for a total of nine time points). Blood is transferred to a 1.5 mL tube pre-spiked with K2EDTA and centrifuged for 4 minutes (8000 rpm, 4°C) to remove plasma. This entire process should be completed within 15 minutes of blood collection. All samples should be stored at -20°C until analysis. The drug concentration was determined by LC / MS / MS. The pharmacokinetic parameters of Compound D, Compound E and the compound of formula (I) in mice at the same dose and administration method are shown in Table 18.

[0351] Table 18 Pharmacokinetic parameters of compounds in mice

[0352] Test Example 6: In vivo efficacy experiment

[0353] In vivo efficacy experiments were performed on BALB / c nude mice subcutaneously implanted with MV4-11 acute myeloid leukemia patient-derived human tumor cell line-based xenografts (CDX).

[0354] Experimental protocol: BALB / c nude mice, female, 6-10 weeks old, weighing approximately 20-23 g, were kept in a special pathogen-free environment in single ventilated cages (5 mice per cage, 10 mice per group, 2 cages). All cages, bedding, and water were disinfected before use. All animals had free access to standard certified commercial laboratory diet. A total of 80 mice purchased from the Laboratory Animal Management Department of Shanghai Institute of Family Planning Science (3577 Jinke Road, Pudong, Shanghai) were used for the study. Each mouse was implanted with tumor cells (1×107 0.1 ml + 0.1 ml Matrigel) was used for tumor growth. When the average tumor volume reached approximately 165 cubic millimeters, the animals were randomly divided into groups based on body weight and tumor volume, and dosing began. The test compound was administered orally by gavage daily. Antitumor efficacy was determined by dividing the average tumor volume increase in animals treated with the compound by the average tumor volume increase in untreated animals.

[0355] Tumor volume was measured twice a week using a two-dimensional caliper and measured in cubic millimeters. Tumor volume TV = 0.5a × b 2 Where a is the long diameter of the tumor and b is the short diameter of the tumor.

[0356] The relative tumor growth rate (T / C) is the percentage of the relative tumor volume (RTV) of the treatment group and the control group at a certain time point. The calculation formula is as follows: T / C% = T RTV / C RTV ×100%(T RTV : Average RTV of treatment group; C RTV : Average RTV of vehicle control group; RTV = V t = / V0, V0 is the tumor volume of the animal at the time of grouping, Vt is the tumor volume of the animal after treatment). mpk refers to milligrams per kilogram of body weight.

[0357] The body weight change (%) of tumor-bearing animals was calculated as follows: (body weight at measurement - body weight at grouping) / body weight at grouping×100.

[0358] The graphs of tumor volume changes and mouse body weight changes after oral administration of compound D and compound of formula (I) once daily are shown in Figures 17 and 18, respectively. The tumor inhibition results of the compounds in mice are shown in Table 19:

[0359] Table 19 Tumor inhibition results of compounds in mice

[0360] All documents mentioned in this disclosure are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of this disclosure, those skilled in the art may make various changes or modifications to this disclosure, and that such equivalents also fall within the scope of the claims appended hereto.

[0361] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0362] The above-described embodiments merely represent several implementation methods of the present disclosure, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the patent. It should be noted that a person of ordinary skill in the art can make a number of variations and improvements without departing from the concept of the present disclosure, and these all fall within the scope of protection of the present disclosure. Therefore, the scope of protection of the patent disclosed herein shall be based on the appended claims, and the description and drawings may be used to interpret the content of the claims.

Claims

1. A pharmaceutically acceptable salt of a compound of formula (I), The pharmaceutically acceptable salt is selected from the group consisting of phosphate and L-tartrate.

2. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 1, wherein The pharmaceutically acceptable salt is L-tartrate; the molar ratio of L-tartaric acid to the compound of formula (I) in the L-tartrate is (0.8-1.2):1, preferably (0.9-1.1):1, and more preferably 1:

1.

3. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 1, wherein The pharmaceutically acceptable salt is a phosphate; the molar ratio of phosphoric acid to the compound of formula (I) in the phosphate is (1.8-2.4):1, preferably (1.9-2.3):1, and more preferably 2:

1.

4. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 1, wherein The pharmaceutically acceptable salt is a phosphate; the molar ratio of phosphoric acid to the compound of formula (I) in the phosphate is (0.8-1.2):1, preferably 1:

1.

5. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 2, wherein The L-tartrate is a polymorph, and the polymorph is selected from the following crystal forms: (1) L-tartrate salt form I, having characteristic diffraction peaks at 2θ (°) diffraction angles of 13.946±0.2, 16.881±0.2, 19.405±0.2, 21.505±0.2, and 24.262±0.2 in its X-ray powder diffraction pattern; and (2) L-tartrate salt form II, whose X-ray powder diffraction pattern has characteristic diffraction peaks at 2θ (°) diffraction angles of 11.662±0.2 and 21.353±0.

2.

6. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 2, wherein The L-tartrate is a polymorph, and the polymorph is selected from the following crystal forms: (1) L-tartrate salt form I, having an X-ray powder diffraction pattern with characteristic diffraction peaks at 2θ (°) diffraction angles of 7.436±0.2, 13.946±0.2, 16.013±0.2, 16.881±0.2, 18.175±0.2, 19.045±0.2, 19.405±0.2, 21.505±0.2, 24.262±0.2, 25.202±0.2, 26.452±0.2, 28.105±0.2, and 31.579±0.2; and (2) L-tartrate salt form II, whose X-ray powder diffraction pattern 2θ (°) diffraction angle has characteristic diffraction peaks at values ​​of 11.662±0.2, 14.244±0.2, 17.481±0.2, 18.349±0.2, 21.026±0.2 and 21.353±0.

2.

7. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 2, wherein The L-tartrate is a polymorph, and the polymorph is selected from the following crystal forms: (1) L-tartrate salt form I, its X-ray powder diffraction pattern 2θ (°) diffraction angle is 6.687±0.2, 7.436±0.2, 10.615±0.2, 12.053±0.2, 13.164±0.2, 13.946±0.2, 14.875±0.2, 15.201±0.2, 16.013±0.2, 16.881±0.2, 18.175±0.2, Characteristic peaks are present at values ​​of 19.045±0.2, 19.405±0.2, 20.659±0.2, 21.505±0.2, 22.434±0.2, 23.04±0.2, 24.262±0.2, 25.202±0.2, 26.452±0.2, 28.105±0.2, 29.692±0.2, 34.139±0.2, and 34.543±0.2; and (2) L-tartrate salt form II, whose X-ray powder diffraction pattern 2θ (°) diffraction angle has characteristic peaks at 10.376±0.2, 11.662±0.2, 14.244±0.2, 16.548±0.2, 17.481±0.2, 18.349±0.2, 18.984±0.2, 21.026±0.2, 21.353±0.2, 26.925±0.2, 29.335±0.2 and 31.784±0.

2.

8. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 2, wherein The L-tartrate is a polymorph, and the polymorph is selected from the following crystal forms: (1) L-tartrate salt Form I having an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG5 ; and (2) L-tartrate salt Form II, which has an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG7 .

9. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 5, wherein The L-tartrate salt Form I has a thermogravimetric analysis (TGA) graph substantially as shown in FIG6 .

10. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 2, wherein The L-tartrate is an amorphous substance.

11. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 3, wherein The phosphate is phosphate crystal form I, and its X-ray powder diffraction pattern 2θ (°) diffraction angle has characteristic diffraction peaks at values ​​of 18.234±0.2, 19.131±0.2 and 21.266±0.

2.

12. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 3, wherein The phosphate is phosphate crystal form I, and its X-ray powder diffraction pattern 2θ (°) diffraction angle has characteristic diffraction peaks at values ​​of 13.999±0.2, 18.234±0.2, 18.803±0.2, 19.131±0.2, 21.266±0.2, 22.01±0.2 and 23.247±0.

2.

13. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 3, wherein The phosphate salt is phosphate salt Form I, which has an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG1 .

14. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 11, wherein The phosphate crystal form I has one or more characteristics selected from the following: having a differential scanning calorimetry (DSC) graph substantially as described in FIG2 ; having a thermogravimetric analysis (TGA) graph substantially as shown in FIG2 ; and There is a dynamic moisture absorption (DVS) diagram substantially as shown in FIG3 .

15. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 4, wherein The phosphate is phosphate crystal form II, and the 2θ (°) diffraction angle of the X-ray powder diffraction pattern thereof has characteristic diffraction peaks at values ​​of 18.230±0.2 and 21.144±0.

2.

16. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 4, wherein The phosphate is phosphate crystal form II, and its X-ray powder diffraction pattern 2θ (°) diffraction angle has characteristic diffraction peaks at values ​​of 10.584±0.2, 13.969±0.2, 14.873±0.2, 18.230±0.2, 20.576±0.2, 21.144±0.2, 22.01±0.2, 22.492±0.2, 23.153±0.2 and 23.96±0.

2.

17. A pharmaceutically acceptable salt of a compound of formula (I) according to claim 4, wherein The phosphate salt is phosphate salt Form II, which has an X-ray powder diffraction (XRPD) pattern substantially as shown in FIG. 10 .

18. A pharmaceutically acceptable salt of a compound of formula (I) according to any one of claims 15 to 17, wherein The phosphate crystal form II has one or more characteristics selected from the following: has a melting temperature of 207.48 ± 0.5°C; having a differential scanning calorimetry (DSC) curve substantially as shown in FIG11 ; having a thermogravimetric analysis (TGA) curve substantially as shown in FIG11 ; and There is a dynamic moisture absorption (DVS) diagram substantially as shown in FIG. 12 .

19. A pharmaceutically acceptable salt of a compound of formula (I) as claimed in any one of claims 15 to 17, wherein The phosphate crystal form II is anhydrous.

20. A method for preparing an L-tartrate salt of a compound of formula (I), comprising the following steps: subjecting the compound of formula (I) to a salt-forming reaction with L-tartaric acid to form the L-tartrate salt of the compound of formula (I).

21. A method for preparing L-tartrate salt form I of the compound of formula (I), comprising the following steps: The compound of formula (I) and L-tartaric acid are subjected to a salt-forming reaction in an organic solvent to form a reaction solution; Slowly cooling the reaction solution to obtain the L-tartrate salt form I; The organic solvent is selected from one or more of ethanol, acetonitrile, ethyl acetate, acetone and methanol.

22. A method for preparing L-tartrate salt form II of the compound of formula (I), comprising the following steps: The compound of formula (I) and L-tartaric acid are subjected to a salt-forming reaction in an organic solvent to form a reaction solution; The reaction solution is slowly cooled, and an anti-solvent is added to obtain the L-tartrate salt form II; wherein the organic solvent is selected from one or more of ethanol, acetonitrile, ethyl acetate, acetone and methanol; The anti-solvent is selected from one or more of methyl tert-butyl ether, petroleum ether, n-heptane, n-hexane, cyclohexane, isopropanol, acetone, acetonitrile and ethyl acetate, and the anti-solvent is different from the organic solvent.

23. The preparation method according to claim 22, wherein The anti-solvent is selected from one or more of methyl tert-butyl ether, n-heptane, isopropanol and acetone, and the anti-solvent is different from the organic solvent.

24. A method for preparing the phosphate according to claim 3 or 4, comprising the steps of: reacting the compound of formula (I) with phosphoric acid in the presence of an organic solvent to form a phosphate of the compound of formula (I).

25. The preparation method according to claim 24, wherein The molar ratio of the compound of formula (I) to phosphoric acid in the phosphate formed is 1:1 or 1:

2.

26. The preparation method according to claim 24, wherein The organic solvent is selected from one or more of ethanol, ethyl acetate, acetonitrile and acetone.

27. The preparation method according to claim 24, wherein The organic solvent is selected from methanol.

28. A method for preparing the phosphate crystal form II of the compound of formula (I), comprising the following steps: The compound of formula (I) is reacted with phosphoric acid in an organic solvent to form a salt to precipitate a solid; The solid was collected to obtain phosphate crystal form II; The molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.8-1.2); the organic solvent is one or more of ethanol, methanol and acetone; preferably, the organic solvent is one or both of ethanol and acetone; preferably, the organic solvent is methanol.

29. The preparation method according to claim 28, wherein The organic solvent is ethanol.

30. The preparation method according to claim 29, wherein The preparation method comprises the following steps: carrying out a salt-forming reaction between an ethanol solution of the compound of formula (I) and an ethanol solution of phosphoric acid to precipitate a solid; The solid was collected to obtain phosphate crystal form II; The molar ratio of the compound of formula (I) to phosphoric acid is 1:(0.8-1.2).

31. A method for preparing the phosphate crystal form I of the compound of formula (I) comprising: (AI-a) dissolving the compound of formula (I) in a solvent, adding an aqueous phosphoric acid solution, and stirring to react; (AI-b) The reaction solution obtained in step (AI-a) is cooled, an anti-solvent is added for crystallization, the solid-liquid separation is performed, and the solid phase is collected to obtain the phosphate crystal form I of the compound of formula (I).

32. A pharmaceutical composition comprising: (a) a pharmaceutically acceptable salt of a compound of formula (I) according to any one of claims 1 to 19; and (b) a pharmaceutically acceptable carrier.

33. Use of a pharmaceutically acceptable salt of a compound of formula (I) according to any one of claims 1 to 19, or a pharmaceutical composition according to claim 32, in the preparation of a medicament for preventing or treating a disease associated with or mediated by CDK9 activity.

34. The use according to claim 33, wherein The disease is one or more of a hyperproliferative disease, a virus-induced infectious disease and a cardiovascular disease.

35. A method for treating a disease associated with or mediated by CDK9 activity, comprising administering to a subject an effective amount of a pharmaceutically acceptable salt of a compound of formula (I) according to any one of claims 1 to 19, or a pharmaceutical composition according to claim 32.

36. A pharmaceutically acceptable salt of a compound of formula (I) according to any one of claims 1 to 19, or a pharmaceutical composition according to claim 32 for use in preventing or treating a disease associated with or mediated by CDK9 activity.

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