Crystal form of KAT6a / b inhibitor, and preparation method therefor and use thereof

Abstract

The present application provides a crystal form of a KAT6A / B inhibitor, and a preparation method therefor and a use thereof. The crystal form provided by the present application has good chemical stability and physical stability, and low moisture absorption, is less affected by heat, humidity, and light, and is convenient for storage and formulation development.

Description

A crystal form of a KAT6A / B inhibitor, its preparation method and application

[0001] This application claims priority to Chinese patent applications 202510002537X, 2025100053308, and 2025119822928, all filed on January 2, 2025. The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This application discloses a crystal form of a KAT6A / B inhibitor, a method for preparing the inhibitor, and their application in the treatment of cancer. Background Technology

[0003] KAT6A (Lysine Acetyltransferase 6A, also known as MOZ) and KAT6B (Lysine Acetyltransferase 6B, also known as MORF) are acetyltransferases belonging to the MYST family. KAT6A exhibits chromosomal translocations in acute myeloid leukemia (AML) and amplification mutations in various cancer types, including lung, breast, ovarian, endometrial, bladder, and esophageal cancers. Similarly, KAT6B also exhibits chromosomal translocation mutations in multiple cancer types. MOZ- and MORF-linked fusion proteins identified in AML include MOZ-CBP, MOZ-p300, MOZ-TIF2, MOZ-NcoA3, MOZ-LEUTX, and MORF-CBP. Among these, MOZ-TIF2 exhibits transforming activity in cultured cells and can induce AML in mice. In tumor cells with KAT6A and KAT6B amplification, the expression of KAT6A and KAT6B is closely related to the gene copy number, indicating that selective pressure exists to maintain their activity during tumorigenesis. Furthermore, in cell proliferation assays, tumor cells with high expression of KAT6A and KAT6B are generally more dependent on the activity of KAT6A and KAT6B.

[0004] Currently, there are many studies based on this mechanism of action, but no KAT6A / B inhibitor drugs have been found on the market. Therefore, there is an urgent need to develop effective KAT6A / B inhibitors for clinical patients. Summary of the Invention

[0005] This application discloses a polymorph of a KAT6A / B inhibitor, a method for preparing the polymorph, and its application in the treatment of cancer.

[0006] Specifically,

[0007] On the one hand, this application provides crystal form A of compound (1aR,7bS)-N-(6-((1H-pyrazol-1-yl)methyl)-4-methoxybenzo[d]isoxazol-3-yl)-5-methoxy-1,1a,2,7b-tetrahydrocyclopropyl[c]chromene-4-sulfonamide, using Cu-Kα irradiation, the X-ray powder diffraction pattern of said crystal form A has characteristic peaks at 2θ values ​​of 12.34°, 15.63°, 18.64°, 20.09°, 21.62°, 22.94°, and 25.38°, with a 2θ error range of ±0.2°;

[0008] In some embodiments of this application, the aforementioned crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 9.51°, 12.34°, 15.63°, 16.72°, 18.64°, 20.09°, 21.42°, 21.62°, 22.61°, 22.94°, 24.28°, 25.38°, 25.71°, 26.26°, 26.72°, and 32.28°, with a 2θ error range of ±0.2°.

[0009] In some embodiments of this application, the aforementioned crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 9.51°, 12.34°, 15.63°, 16.77°, 18.64°, 20.09°, 21.42°, 21.62°, 22.61°, 22.94°, 24.28°, 25.38°, 25.71°, 26.26°, 26.72°, and 32.28°, with a 2θ error range of ±0.2°.

[0010] In some embodiments of this application, the aforementioned crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 9.51°, 11.19°, 11.79°, 12.34°, 12.71°, 13.22°, 15.18°, 15.63°, 16.72°, 18.64°, 19.20°, 20.09°, 21.42°, 21.62°, 22.61°, 22.94°, 24.28°, 25.38°, 25.71°, 26.26°, 26.72°, 28.9°, 30.10°, 31.44°, 32.28°, and 37.68°, with a 2θ error range of ±0.2°.

[0011] In some embodiments of this application, the aforementioned crystal form A, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 9.51°, 11.19°, 11.79°, 12.34°, 12.71°, 13.22°, 15.18°, 15.63°, 16.77°, 18.64°, 19.20°, 20.09°, 21.42°, 21.62°, 22.61°, 22.94°, 24.28°, 25.38°, 25.71°, 26.26°, 26.72°, 28.9°, 30.10°, 31.44°, 32.28°, and 37.68°, with a 2θ error range of ±0.2°.

[0012] In some embodiments of this application, the X-ray powder diffraction pattern of the crystal form A is shown in Figure 3, with a 2θ error range of ±0.2°.

[0013] In some embodiments of this application, the TGA-DSC spectrum of the aforementioned crystal form A shows a weight loss of 29.59% at 35.46±3 to 351.51±3 °C, preferably at 35.46±2 to 351.51±2 °C, for example at 35.46 to 351.51 °C.

[0014] In some embodiments of this application, the TGA-DSC spectrum of the aforementioned crystal form A has an endothermic peak at 197.75±5℃ and an exothermic peak at 268.77±5℃. Preferably, it has an endothermic peak at 197.75±2℃ and an exothermic peak at 268.77±2℃. For example, it has an endothermic peak at 197.75℃ and an exothermic peak at 268.77℃.

[0015] In some embodiments of this application, the crystal form A described above has a TGA-DSC spectrum as shown in Figure 4.

[0016] In some embodiments of the present invention, the crystal of the above crystal form A is an orthorhombic crystal system with space group P22121 and cell parameters a = 8.0814(6)A, b = 14.8372(11), c = 18.3332(14).

[0017] In some embodiments of the present invention, the single crystal structure spectrum of the above-mentioned crystal form A is shown in Figure 2.

[0018] In some embodiments of this application, the XRPD diffraction peak analysis data of crystal form A of the above formula (Ⅰ) are shown in Table 1.

[0019] Table 1. XRPD diffraction peak analysis data for crystal form A of compound (I)

[0020] On the other hand, the present invention provides p-toluenesulfonate of compound (I):

[0021] The molar ratio of free base to p-toluenesulfonic acid is 1:1.

[0022] Specifically, the present invention provides p-toluenesulfonate crystal form I of compound (I), which, when irradiated with Cu-Kα, has characteristic peaks at 2θ values ​​of 11.20, 12.80, 15.75, 16.12, 20.17, and 24.32, with a 2θ error range of ±0.2°.

[0023] In some embodiments of the present invention, the p-toluenesulfonate crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 8.04, 11.20, 12.40, 12.80, 13.72, 15.75, 16.12, 20.17, 20.92, 21.93, 24.32, and 25.15, with a 2θ error range of ±0.2°.

[0024] In some embodiments of the present invention, the p-toluenesulfonate crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 8.04, 11.20, 11.94, 12.40, 12.80, 13.72, 15.30, 15.75, 16.12, 18.42, 18.74, 19.42, 20.17, 20.92, 21.93, 22.32, 22.74, 23.51, 24.32, and 25.15, with a 2θ error range of ±0.2°.

[0025] In some embodiments of the present invention, the p-toluenesulfonate crystal form I of the above-mentioned formula (Ⅰ) compound has an X-ray powder diffraction pattern as shown in Figure 5, with a 2θ error range of ±0.2°.

[0026] In some embodiments of the present invention, the p-toluenesulfonate crystal form I of the above-mentioned formula (I) has a TGA-DSC spectrum showing a weight loss of 33.29% at 35.61±3 to 351.92±3 °C, preferably at 35.61±2 to 351.92±2 °C, for example at 35.61 to 351.92 °C.

[0027] In some embodiments of the present invention, the p-toluenesulfonate crystal form I of the above-mentioned formula (I) has an endothermic peak at 145.85±5℃ in its TGA-DSC spectrum, preferably an endothermic peak at 145.85±2℃, for example, an endothermic peak at 145.85℃.

[0028] In some embodiments of the present invention, the p-toluenesulfonate crystal form I of the above-mentioned formula (I) is shown in Figure 6.

[0029] In some embodiments of the present invention, the XRPD diffraction peak analysis data of the p-toluenesulfonate crystal form I of the above-mentioned formula (Ⅰ) compound are shown in Table 2.

[0030] Table 2 shows the XRPD diffraction peak analysis data of p-toluenesulfonate crystal form I of compound (I).

[0031] On the other hand, the present invention provides a potassium salt crystal form II of the compound of formula (I), which, when irradiated with Cu-Kα, has characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 9.85, 15.32, 19.44, 20.50, 20.86, and 22.81, with a 2θ error range of ±0.2°.

[0032] In some embodiments of the present invention, the potassium salt crystal form II of the above-mentioned compound (I) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 8.86, 9.85, 10.93, 14.70, 15.32, 16.54, 19.44, 20.50, 20.86, 22.81, 25.67, and 27.06, with a 2θ error range of ±0.2°.

[0033] In some embodiments of the present invention, the potassium salt crystal form II of the above-mentioned compound (I) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 8.86, 9.85, 10.93, 12.25, 12.90, 13.88, 14.70, 15.32, 16.54, 17.85, 19.44, 20.50, 20.86, 21.77, 22.81, 25.67, 27.06, 27.63, 31.37, 32.77, and 34.21, with a 2θ error range of ±0.2°.

[0034] In some embodiments of the present invention, the potassium salt crystal form II of the above-mentioned formula (I) compound has an X-ray powder diffraction pattern as shown in Figure 7, with a 2θ error range of ±0.2°.

[0035] In some embodiments of the present invention, the potassium salt crystal form II of the above-mentioned formula (I) compound exhibits a weight loss of 21.97% in its TGA-DSC spectrum at 35.56±3 to 352.05±3 °C, preferably at 35.56±2 to 352.05±2 °C, for example at 35.56 to 352.05 °C.

[0036] In some embodiments of the present invention, the potassium salt crystal form II of the above-mentioned formula (I) compound has an exothermic peak at 260.64℃±5℃ in its TGA-DSC spectrum, preferably at 260.64℃±2℃, for example, at 260.64℃.

[0037] In some embodiments of the present invention, the potassium salt crystal form II of the above-mentioned formula (I) compound is shown in Figure 8.

[0038] In some embodiments of the present invention, the XRPD diffraction peak analysis data of the potassium salt crystal form II of the above-mentioned formula (I) compound are shown in Table 3.

[0039] Table 3. XRPD diffraction peak analysis data of potassium salt crystal form II of compound (I).

[0040] On the other hand, the present invention provides a hydrochloride crystal form I of the compound of formula (Ⅰ).

[0041] The molar ratio of free base to hydrochloric acid is 1:1.

[0042] In some embodiments of the present invention, the hydrochloride crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and the X-ray powder diffraction pattern of the hydrochloride crystal form I has characteristic peaks at 2θ values ​​of 11.65, 19.68, 21.06, 22.65, 22.92, and 23.27, with a 2θ error range of ±0.2°.

[0043] In some embodiments of the present invention, the hydrochloride crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 11.08, 11.65, 12.11, 12.31, 14.30, 19.68, 20.07, 20.54, 21.06, 22.65, 22.92, and 23.27, with a 2θ error range of ±0.2°.

[0044] In some embodiments of the present invention, the hydrochloride crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 6.77, 7.53, 11.08, 11.65, 12.11, 12.31, 14.30, 15.40, 17.20, 17.82, 18.15, 19.68, 20.07, 20.54, 21.06, 22.12, 22.65, 22.92, 23.27, 23.54, 25.17, and 29.60, with a 2θ error range of ±0.2°.

[0045] In some embodiments of the present invention, the X-ray powder diffraction pattern of the hydrochloride crystal form I of the above-mentioned formula (Ⅰ) compound is shown in Figure 9, with a 2θ error range of ±0.2°.

[0046] In some embodiments of the present invention, the hydrochloride crystal form I of the above-mentioned formula (I) compound exhibits a TGA-DSC spectrum showing a weight loss of 6.86% at 35.68±3 to 209.55±3 °C and a weight loss of 25.39% at 209.55±3 to 352.04±3 °C. Preferably, the weight loss is 6.86% at 35.68±2 to 209.55±2 °C and a weight loss of 25.39% at 209.55±2 to 352.04±2 °C. For example, the weight loss is 6.86% at 35.68 to 209.55 °C and a weight loss of 25.39% at 209.55 to 352.04 °C.

[0047] In some embodiments of the present invention, the hydrochloride crystal form I of the compound of formula (I) above has an endothermic peak at 157.04±5℃ and an endothermic peak at 197.45±5℃ in its TGA-DSC spectrum. Preferably, it has an endothermic peak at 157.04±2℃ and an endothermic peak at 197.45±2℃. For example, it has an endothermic peak at 157.04℃ and an endothermic peak at 197.45℃.

[0048] In some embodiments of the present invention, the hydrochloride crystal form I of the compound of formula (I) above, the TGA-DSC spectrum of which is shown in Figure 10.

[0049] In some embodiments of the present invention, the XRPD diffraction peak analysis data of the hydrochloride crystal form I of the above-mentioned formula (Ⅰ) compound are shown in Table 4.

[0050] Table 4. XRPD diffraction peak analysis data of hydrochloride crystal form I of compound (I)

[0051] On the other hand, the present invention provides a methanesulfonate crystal form I of the compound of formula (Ⅰ).

[0052] The molar ratio of free base to methanesulfonic acid is 1:1.6.

[0053] In some embodiments of the present invention, the methanesulfonate crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 6.89, 13.77, 18.02, 21.81, 23.67, and 24.67, with a 2θ error range of ±0.2°.

[0054] In some embodiments of the present invention, the methanesulfonate crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 6.89, 9.55, 12.24, 13.77, 16.74, 18.02, 19.03, 21.33, 21.81, 22.34, 23.67, and 24.67, with a 2θ error range of ±0.2°.

[0055] In some embodiments of the present invention, the methanesulfonate crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 6.89, 9.55, 10.35, 10.74, 12.24, 12.59, 13.77, 16.50, 16.74, 18.02, 19.03, 20.78, 21.33, 21.81, 22.34, 23.67, 24.67, 25.21, 26.29, 26.67, and 27.70, with a 2θ error range of ±0.2°.

[0056] In some embodiments of the present invention, the methanesulfonate crystal form I of the above-mentioned formula (Ⅰ) compound has an X-ray powder diffraction pattern as shown in Figure 11, with a 2θ error range of ±0.2°.

[0057] In some embodiments of the present invention, the methanesulfonate crystal form I of the above-mentioned formula (I) compound exhibits a weight loss of 31.67% in its TGA-DSC spectrum at 35.62±3 to 351.69±3 °C, preferably at 35.62±2 to 351.69±2 °C, for example at 35.62 to 351.69 °C.

[0058] In some embodiments of the present invention, the methanesulfonate crystal form I of the above-mentioned formula (I) has an endothermic peak at 105.05±5℃ in its TGA-DSC spectrum, preferably an endothermic peak at 105.05±2℃, for example, an endothermic peak at 105.05℃.

[0059] In some embodiments of the present invention, the methanesulfonate crystal form I of the above-mentioned formula (I) compound is shown in Figure 12.

[0060] In some embodiments of the present invention, the XRPD diffraction peak analysis data of the methanesulfonate crystal form I of the above-mentioned formula (Ⅰ) compound are shown in Table 5.

[0061] Table 5. XRPD diffraction peak analysis data of methanesulfonate crystal form I of compound (Ⅰ).

[0062] On the other hand, the present invention provides a potassium salt crystal form I of the compound of formula (Ⅰ), which, when irradiated with Cu-Kα, has characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 9.49, 10.20, 15.40, 18.09, 22.65, and 22.97, with a 2θ error range of ±0.2°.

[0063] In some embodiments of the present invention, the potassium salt crystal form I of the above-mentioned (Ⅰ) compound, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 9.49, 10.20, 10.90, 12.35, 15.40, 17.57, 18.09, 21.67, 22.65, 22.97, 23.21, and 23.94, with a 2θ error range of ±0.2°.

[0064] In some embodiments of the present invention, the potassium salt crystal form I of the above-mentioned (Ⅰ) compound, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its X-ray powder diffraction pattern at 2θ values ​​of 8.29, 9.49, 10.20, 10.90, 11.83, 12.35, 12.90, 13.48, 13.95, 15.40, 17.57, 18.09, 20.10, 20.49, 21.67, 22.65, 22.97, 23.21, 23.94, 24.88, and 27.80, with a 2θ error range of ±0.2°.

[0065] In some embodiments of the present invention, the potassium salt crystal form I of the above (Ⅰ) compound has an X-ray powder diffraction pattern as shown in Figure 13, with a 2θ error range of ±0.2°.

[0066] In some embodiments of the present invention, the potassium salt crystal form I of the above-mentioned formula (I) compound exhibits a weight loss of 1.54% at 35.75±3 to 210.78±3 °C and a weight loss of 21.28% at 210.78±3 to 351.96±3 °C. Preferably, the weight loss is 1.54% at 35.75±2 to 210.78±2 °C and a weight loss of 21.28% at 210.78±2 to 351.96±2 °C. For example, the weight loss is 1.54% at 35.75 to 210.78 °C and a weight loss of 21.28% at 210.78 to 351.96 °C.

[0067] In some embodiments of the present invention, the potassium salt crystal form I of the above-mentioned formula (I) compound has a TGA-DSC spectrum showing an endothermic peak at 175.36℃±5℃, an exothermic peak at 217.41±5℃, and an exothermic peak at 259.80±5℃. Preferably, it has an endothermic peak at 175.36℃±2℃, an exothermic peak at 217.41±2℃, and an exothermic peak at 259.80±2℃. For example, it has an endothermic peak at 175.36℃, an exothermic peak at 217.41℃, and an exothermic peak at 259.80℃.

[0068] In some embodiments of the present invention, the potassium salt crystal form I of the above (Ⅰ) compound is shown in Figure 14.

[0069] In some embodiments of the present invention, the XRPD diffraction peak analysis data of the potassium salt crystal form I of the above-mentioned formula (Ⅰ) compound are shown in Table 6.

[0070] Table 6. XRPD diffraction peak analysis data of potassium salt crystal form I of compound (Ⅰ).

[0071] On the other hand, the present invention provides choline salt crystal form I of compound (Ⅰ).

[0072] The molar ratio of free base to choline is 1:1.

[0073] In some embodiments of the present invention, the choline salt crystal form I of the above-mentioned formula (Ⅰ) compound is subjected to Cu-Kα radiation, and the X-ray powder diffraction pattern of the crystal form I has characteristic peaks at 2θ values ​​of 9.41, 14.21, 16.37, 18.98, 21.67, and 22.56, with a 2θ error range of ±0.2°.

[0074] In some embodiments of the present invention, the choline salt crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 9.41, 12.25, 12.78, 14.21, 14.39, 16.37, 17.94, 18.98, 20.99, 21.44, 21.67, and 22.56, with a 2θ error range of ±0.2°.

[0075] In some embodiments of the present invention, the choline salt crystal form I of the above-mentioned compound (Ⅰ) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.06, 9.41, 12.25, 12.78, 14.21, 14.39, 16.37, 17.94, 18.98, 20.99, 21.44, 21.67, 21.89, 22.56, 23.52, 24.22, 24.41, 24.73, 25.01, and 27.06, with a 2θ error range of ±0.2°.

[0076] In some embodiments of the present invention, the choline salt crystal form I of the above-mentioned formula (Ⅰ) compound has an X-ray powder diffraction pattern as shown in Figure 15, with a 2θ error range of ±0.2°.

[0077] In some embodiments of the present invention, the choline salt crystal form I of the above-mentioned formula (I) has a TGA-DSC spectrum showing a weight loss of 2.01% at 35.55±3 to 127.39±3 °C and a weight loss of 36.66% at 127.39±3 to 351.97±3 °C. Preferably, it shows a weight loss of 2.01% at 35.55±2 to 127.39±2 °C and a weight loss of 36.66% at 127.39±2 to 351.97±2 °C. For example, it shows a weight loss of 2.01% at 35.55 to 127.39 °C and a weight loss of 36.66% at 127.39 to 351.97 °C.

[0078] In some embodiments of the present invention, the choline salt crystal form I of the above-mentioned formula (I) has an endothermic peak at 101.67±5℃ in its TGA-DSC spectrum, preferably an endothermic peak at 101.67±2℃, for example, an endothermic peak at 101.67℃.

[0079] In some embodiments of the present invention, the choline salt crystal form I of the above-mentioned formula (Ⅰ) compound is shown in Figure 16.

[0080] In some embodiments of the present invention, the XRPD diffraction peak analysis data of the choline salt crystal form I of the above formula (Ⅰ) compound are shown in Table 7.

[0081] Table 7. XRPD diffraction peak analysis data of choline salt crystal form I of compound (Ⅰ).

[0082] On the other hand, this application also provides a pharmaceutical composition comprising any of the above-mentioned crystal forms and a pharmaceutically acceptable carrier.

[0083] On the other hand, this application also provides a pharmaceutical composition comprising any of the above-mentioned salt forms and a pharmaceutically acceptable carrier.

[0084] On the other hand, this application also provides the use of the above-mentioned arbitrary crystal form and pharmaceutical composition in the preparation of a drug for treating KAT6-mediated cancer.

[0085] On the other hand, this application also provides the use of the above-mentioned salt forms and pharmaceutical compositions in the preparation of drugs for treating KAT6-mediated cancer.

[0086] On the other hand, this application also provides the use of the above-mentioned crystal form and pharmaceutical composition for treating KAT6A / B-mediated cancers.

[0087] On the other hand, this application also provides the use of the above-mentioned salt forms and pharmaceutical compositions for treating KAT6A / B-mediated cancers.

[0088] In some embodiments of this application, the aforementioned cancers include breast cancer, prostate cancer, and lung cancer.

[0089] On the other hand, this application also provides a method for preparing crystal form A, including the following steps:

[0090] (a) Dissolve the compound of formula (Ⅰ) in a first solvent, heat, stir, filter, and wash;

[0091] (b) At a certain temperature, add the second solvent to the solution in step (a) and stir for a certain time;

[0092] (c) Cool down, stir for a certain time, filter, wash, collect, and dry the solid.

[0093] In some embodiments of this application, the first solvent in the above preparation method is selected from one or a mixture of two of dichloromethane and dimethyl sulfoxide; preferably dimethyl sulfoxide.

[0094] In some embodiments of this application, the second solvent in the above preparation method is selected from one or more mixtures of acetone, butanone, methyl isobutyl ketone, methanol, ethanol, isopropanol, ethyl acetate, isopropyl acetate, tetrahydrofuran, 1,4-dioxane, methyl tert-butyl ether, water, and acetonitrile; ethanol is preferred.

[0095] In some embodiments of this application, in step (a) of the above preparation method, the amount of the first solvent and the endpoint temperature of the heating are not limited, the purpose of which is to dissolve the solid.

[0096] In some embodiments of this application, in step (a) of the above preparation method, the volume-to-mass ratio of the first solvent to the compound of formula (I) is preferably 3.5 to 5.5 mL / g, for example 4 mL / g or 5 mL / g. The endpoint temperature of the heating is preferably 25 to 90°C, more preferably 30 to 35°C or 80 to 85°C.

[0097] In some embodiments of this application, the filtration in the above preparation method can be a conventional filtration method in the art, preferably gravity filtration or vacuum filtration.

[0098] In some embodiments of this application, in step (a) of the preparation method described above, the solvent used for washing is a first solvent, wherein the amount of the first solvent can be the amount of washing solvent conventional in the art, preferably an amount with a volume-to-mass ratio of 0.5 to 5.5 mL / g to the compound of formula (I), for example, an amount with a volume-to-mass ratio of 1 mL / g or 5 mL / g to the compound of formula (I).

[0099] In some embodiments of this application, in step (b) of the above preparation method, the temperature can be a conventional mixing temperature in the art, preferably 25 to 65°C, more preferably 30 to 35°C or 55 to 60°C. The amount of the second solvent can be a conventional solvent amount in the art, preferably an amount with a volume-to-mass ratio of 5 to 105 mL / g of the compound of formula (I), for example, an amount with a volume-to-mass ratio of 10 mL / g, 20 mL / g or 100 mL / g of the compound of formula (I).

[0100] In some embodiments of this application, in step (b) of the above preparation method, the stirring time can be a conventional stirring time in the art, preferably 50±10 to 70±10 min, for example 60±10 min.

[0101] In some embodiments of this application, in step (c) of the above preparation method, the endpoint temperature of the cooling can be a conventional endpoint temperature in the art, preferably -0.5±5 to 1±5℃, for example 0±5℃.

[0102] In some embodiments of this application, in step (c) of the above preparation method, the stirring time can be a conventional stirring time in the art, preferably 1.5±1 to 2.5±1 h, for example 2±1 h.

[0103] In some embodiments of this application, in step (c) of the preparation method described above, the solvent used for washing is a second solvent, wherein the amount of the second solvent can be the amount of washing solvent conventional in the art, preferably an amount with a volume-to-mass ratio of 2 to 3 mL / g to the compound of formula (I), for example, an amount with a volume-to-mass ratio of 2.86 mL / g to the compound of formula (I).

[0104] In some embodiments of this application, in step (c) of the above preparation method, the drying can be a conventional drying method in the art, preferably vacuum drying. The vacuum drying temperature can be a conventional vacuum drying temperature in the art, preferably 65±5 to 75±5℃, for example 70±5℃. The vacuum drying time can be a conventional vacuum drying time in the art, preferably 5 to 7 hours, for example 6 hours.

[0105] Technical effect

[0106] The crystal and salt forms in this application have good chemical and physical stability and low hygroscopicity. They are less affected by temperature, humidity and light, making them easy to store and develop formulations.

[0107] Definitions and Explanations

[0108] Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense.

[0109] The term "pharmaceutical-grade carrier" refers to a medium generally acceptable in the art for delivering a bioactive pharmaceutical agent to animals, particularly mammals. Depending on the route of administration and dosage form, this includes, for example, adjuvants, excipients, or excipients such as diluents, preservatives, fillers, flow modifiers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatizers, antibacterial agents, antifungal agents, lubricants, and dispersants. Pharmaceutically acceptable carriers are formulated based on a multitude of factors, within the scope of those skilled in the art. These include, but are not limited to, the type and nature of the active pharmaceutical agent being formulated, the recipient to whom the composition containing the pharmaceutical agent is to be administered, the intended route of administration of the composition, and the target therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous media, as well as various solid and semi-solid dosage forms. In addition to the active pharmaceutical agent, such carriers include many different components and additives, and the inclusion of such additional components in the formulation for various reasons (e.g., stabilizing active pharmaceutical agents, binders, etc.) is well known to those skilled in the art.

[0110] As is known in the art, X-ray powder diffraction patterns have one or more measurement errors due to minute variations in measurement conditions. The structures of crystals, crystal forms, or crystalline forms disclosed or claimed in this application may exhibit similar but not identical analytical characteristics within a reasonable error range depending on experimental conditions, purity, equipment, and other constant variables known to those skilled in the art. For example, the diffraction angle (2θ) in powder X-ray powder diffraction typically produces an error within ±0.20°. Therefore, this application includes not only crystals with completely consistent diffraction angles in powder X-ray powder diffraction, but also crystals with consistent diffraction angles within an error range of ±0.20°. The crystalline form of the compound of formula (I) in this application is not limited to crystals having the same X-ray powder diffraction pattern as shown in the accompanying drawings; any crystal having a substantially identical X-ray powder diffraction pattern as shown in the accompanying drawings falls within the scope of this application. In this invention, the XRPD pattern testing conditions preferably include, in one embodiment, a Cu target as the light source. In another embodiment, the scanning method is a single scan. In one scheme, the scanning angle is 3-45°. In another scheme, the scanning step size is 0.02°. In yet another scheme, the time per step is 40 seconds.

[0111] The text refers to "X-ray powder diffraction patterns that are substantially the same as those shown in the accompanying figures." It should be understood that the term "substantially the same" used in this context also indicates that the 2θ angle values ​​of the X-ray powder diffraction patterns may vary slightly due to inherent experimental variations accompanying these measurements, and both are of the same crystalline form.

[0112] It should be understood that different types of equipment or different testing conditions may yield slightly different DSC spectra and endothermic transition temperature readings. DSC data can reflect changes in the state of matter; strong endothermic peaks can indicate that the substance has undergone dehydration or desolvation, or crystal transformation, or melting, etc.; when reflecting the molten state, the corresponding temperature is generally understood as the melting point of the substance. This value will be affected by the purity of the compound, sample weight, heating rate, particle size, and calibration and maintenance of the testing equipment. Those skilled in the art will understand that the temperature at which a substance changes from a solid to a liquid state is usually a temperature range, not a fixed point value; therefore, the temperature corresponding to the endothermic peak or the melting point of the substance can be characterized by either the onset value, the peak value, or other reasonable values. The maximum endothermic transition temperature of the crystal form can be within the range of ±5.0℃ of the specific values ​​disclosed above, preferably within the range of ±2.0℃. For example, an endothermic peak at around 253.5℃ in the DSC spectrum means an endothermic peak at 253.5℃ ±5.0℃, preferably 253.5℃ ±2.0℃.

[0113] This application also uses thermogravimetric analysis (TGA) to analyze the relationship between the degree of decomposition, sublimation, or evaporation of the crystal form (weight loss) and temperature. It should be understood that the values ​​obtained for the same crystal form may have some error due to factors such as sample purity, particle size, different types of equipment, and different testing methods. The temperature at which the crystal form decomposes, sublimates, or evaporates can be within ±3.0℃ of the specific values ​​disclosed above, for example, within ±2.0℃. In this invention, the testing conditions for the TGA-DSC spectrum are preferably as follows: in one scheme, the temperature range is room temperature to 400℃; in another scheme, the heating rate is 10℃ / min; in yet another scheme, the flow rate in the equilibrium chamber is 20 mL / min; and in yet another scheme, the flow rate in the sample chamber is 50 mL / min.

[0114] The "stability" of a crystal form includes "chemical stability" and / or "physical stability." "Chemical stability" refers to the degree to which the crystal form undergoes degradation reactions under certain temperature, humidity, and light conditions; it reflects the stability of the crystal form under storage conditions. "Physical stability" refers to the degree to which the crystal form undergoes a solid-state transformation under certain specific conditions, such as under conditions of high temperature, high humidity, grinding, tableting, solvent removal, or solvent adsorption, transforming into another crystal form. Therefore, "physical stability" can, to some extent, reflect the stability of the crystal form during its use in formulations and other processes.

[0115] The crystalline structure of this application can be prepared by various methods, including crystallization or recrystallization from a suitable solvent, sublimation, growth from a melt, solid-state transformation from another phase, crystallization from a supercritical fluid, and jet spraying. Techniques for crystallizing or recrystallizing the crystalline structure from a solvent mixture include solvent evaporation, lowering the temperature of the solvent mixture, crystallization of a supersaturated solvent mixture of the molecule and / or salt, lyophilizing the solvent mixture, and adding an antisolvent to the solvent mixture.

[0116] The term "drying" refers to the process of removing solvent from the resulting solid, including but not limited to air drying at room temperature, high-temperature drying, and vacuum drying.

[0117] The intermediate compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application.

[0118] In the embodiments of this application, if there is a discrepancy between the compound name and the compound structure, the discrepancy can be determined by combining relevant information and reaction routes; if it cannot be confirmed by other means, the given compound structural formula shall prevail.

[0119] The preparation methods for some compounds in this application reference the preparation methods for the aforementioned similar compounds. Those skilled in the art should understand that when using or referring to the referenced preparation methods, the reactant ratios, reaction solvents, reaction temperatures, etc., can be appropriately adjusted according to the different reactants.

[0120] The compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application.

[0121] Instruments and analytical methods:

[0122] 1. X-ray powder diffraction (XRPD)

[0123] Solid samples were analyzed using an X-ray powder diffractometer (Aeris). An appropriate amount of fine powder was taken and placed in the groove of the sample holder. A glass slide was used to press the powder into a flat and dense plane. The XRPD measurement parameters are shown in Table 8.

[0124] Table 8 XRPD Test Parameters

[0125] 2. Simultaneous Thermal Analysis (TGA-DSC)

[0126] Alternatively, a Mettler Toledo simultaneous thermal analyzer can be used for thermogravimetric-differential scanning calorimetry (TGC) analysis of solids. Take an appropriate amount of the sample with a small spoon and place it in a crucible, spreading it evenly. Weigh the sample and heat it according to the parameters listed in Table 9. Analyze the data using STARE.

[0127] Table 9 Parameters of TGA-DSC Analysis Method

[0128] 3. Dynamic moisture adsorption-desorption analysis (DVS)

[0129] The hygroscopicity of the samples was determined using a DVS Intrinsic dynamic moisture adsorption analyzer. The samples were placed in a pre-peeled sample basket, and the instrument automatically weighed them. The samples were then analyzed according to the parameters in Table 10.

[0130] Table 10 Parameters of DVS Analysis Method Attached Figure Description

[0131] Figure 1 shows the amorphous spectrum of compound (Ⅰ).

[0132] Figure 2 shows the single-crystal diffraction pattern of compound (Ⅰ).

[0133] Figure 3 shows the AX-ray powder diffraction pattern of the crystal form of compound (Ⅰ).

[0134] Figure 4 shows the ATGA-DSC spectrum of the crystal form of compound (Ⅰ).

[0135] Figure 5 shows the X-ray powder diffraction pattern of p-toluenesulfonate crystal form I of compound (Ⅰ).

[0136] Figure 6 shows the TGA-DSC spectrum of p-toluenesulfonate of compound (Ⅰ).

[0137] Figure 7 shows the X-ray powder diffraction pattern of potassium salt crystal form II of compound (Ⅰ).

[0138] Figure 8 shows the TGA-DSC spectrum of the potassium salt of compound (Ⅰ) in crystal form Ⅱ.

[0139] Figure 9 shows the X-ray powder diffraction pattern of the hydrochloride crystal form I of compound (Ⅰ).

[0140] Figure 10 shows the TGA-DSC spectrum of the hydrochloride crystal form of compound (Ⅰ).

[0141] Figure 11 shows the X-ray powder diffraction pattern of the methanesulfonate crystal form I of compound (Ⅰ).

[0142] Figure 12 shows the TGA-DSC spectrum of the methanesulfonate crystal form of compound (Ⅰ).

[0143] Figure 13 shows the X-ray powder diffraction pattern of potassium salt crystal form I of compound (Ⅰ).

[0144] Figure 14 shows the TGA-DSC spectrum of the potassium salt crystal form of compound (Ⅰ).

[0145] Figure 15 shows the X-ray powder diffraction pattern of choline salt crystal form I of compound (Ⅰ).

[0146] Figure 16 shows the TGA-DSC spectrum of choline salt crystal form I of compound (Ⅰ).

[0147] Figure 17 shows the changes in body weight of mice in groups G1 to G4 in Test Example 3 of Example 2 of this application.

[0148] Figure 18 shows the tumor growth in mice in groups G1 to G4 of Example 3 of this application.

[0149] Figure 19. DVS spectrum of crystal form A. Detailed Implementation

[0150] The present application is described in detail below through examples, but this does not imply any adverse limitation on the present application. The compounds of this application can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of this application without departing from the spirit and scope of this application.

[0151] Detailed implementation method:

[0152] Intermediate INT-1: ((1aR,7bS)-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene-4-sulfonyl chloride)

[0153] Reaction route:

[0154] Step 1: Preparation of intermediate a ((1aR,7bS)-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene)

[0155] At room temperature, sodium metal (7.1 g, 310 mmol) was dissolved in anhydrous diethyl ether (70 mL), and 1,1-dichloro-5-methoxy-2,7b-dihydro-1aH-cyclopropyl[c]chromene (9.5 g, 38.8 mmol) was dissolved in a mixture of anhydrous diethyl ether (30 mL) and anhydrous methanol (15 mL). The solutions were slowly added dropwise to the reaction mixture, with 5 mL of methanol added every hour until the starting materials disappeared.

[0156] After LCMS monitoring showed the disappearance of the starting material, the reaction was quenched with anhydrous ethanol (20 mL), diluted with water (50 mL), and extracted with ethyl acetate (50 mL × 3). The organic phase was washed with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by rapid silica gel column chromatography to obtain 5.5 g of 5-methoxy-1,1a,2,7b-tetrahydrocyclopropyl[c]benzopyran racemate; the racemate was separated by SFC (column: DAICEL CHIRALCEL OJ (250 mm * 50 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 20%%, isocrine mode) to obtain (1aR,7bS)-5-methoxy-1,1a,2,7b-tetrahydrocyclopropyl[c]chromene.

[0157] 1H NMR(400MHz,CHLOROFORM-d)δ0.92-1.06(m,2H),1.66-1.75(m,1H),1.93(td,J=8.4,4.5Hz,1H),3.76(s,3H),3.91( dd,J=10.5,1.3Hz,1H),4.27-4.35(m,1H),6.41(d,J=2.5Hz,1H),6.50(dd,J=8.3,2.5Hz,1H),7.13(d,J=8.3Hz,1H).

[0158] Step 2: Preparation of intermediate b ((1aR,7bS)-4-benzylsulfonyl-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene)

[0159] The starting material (1aR,7bS)-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene (600 mg, 3.4 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), and nitrogen was purged three times. Then, n-butyllithium (1.6 mL, 4.1 mmol) was added dropwise under ice bath conditions and stirred for 1 hour. Then, 1,2-dibenzyl disulfide (1.0 g, 4.1 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and added dropwise to the reaction solution. The mixture was gradually heated to room temperature and reacted for 2 hours.

[0160] After LCMS monitoring showed the starting material had disappeared, water (30 mL) was added to dilute the reaction system. The mixture was extracted with ethyl acetate (30 mL × 3), and the organic phases were combined. The organic phase was first washed with saturated brine (30 mL × 3), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by rapid silica gel column chromatography to obtain 250 mg of (1aR,7bS)-4-benzylsulfonyl-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene.

[0161] 1 H NMR(400MHz,CHLOROFORM-d)δppm 0.87-0.98(m,2H),1.61(br s,1H),1.88(td,J=8.3,4.6Hz,1H),3.51-3.58(m,1H),3.78-3.84(m,3H),3.87-3.94(m,1 H),3.97-4.03(m,1H),4.33(dd,J=10.4,0.9Hz,1H),6.43-6.49(m,1H),7.08-7.23(m,6H).

[0162] Step 3: Preparation of intermediate INT-1 ((1aR,7bS)-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene-4-sulfonyl chloride)

[0163] (1aR,7bS)-4-benzylsulfonyl-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene (250 mg, 0.84 mmol) was dissolved in acetonitrile (5 mL) and water (0.5 mL), acetic acid (403 mg, 6.7 mmol) was added, and the mixture was cooled in an ice bath. Then, dichlorohydantoin (331 mg, 1.7 mmol) was added to the reaction solution, and the mixture was reacted in an ice bath for half an hour.

[0164] After thin-layer chromatography monitoring showed the starting material had disappeared, water (30 mL) was added to dilute the reaction system. The mixture was extracted with ethyl acetate (30 mL × 3), and the organic phases were combined. The organic phase was first washed with saturated brine (30 mL × 3), then dried over anhydrous sodium sulfate, filtered, and finally concentrated under reduced pressure. The crude product was purified by rapid silica gel column chromatography to give 100 mg of (1aR,7bS)-5-methoxy-1,1a,2,7b-tetrahydrocyclopropane[c]chromene-4-sulfonyl chloride.

[0165] 1 H NMR(400MHz,CHLOROFORM-d)δppm 1.04-1.14(m,2H),1.75-1.84(m,1H),1.93-2.04(m,1H),3.92-3.96(m,3H),4.00(d d,J=10.7,1.9Hz,1H),4.56(d,J=10.5Hz,1H),6.57-6.65(m,1H),7.42-7.50(m,1H).

[0166] Example 1: Preparation of compound (I)

[0167] 12.9 g (0.047 mol) of intermediate INT-1 and 10 g (0.04 mol) of 6-((1H-pyrazol-1-yl)methyl)-4-methoxybenzo[d]isoxazole-3-amine were added to 29.7 g of pyridine and the mixture was heated to 80±5 °C and reacted for 4 h.

[0168] After the reaction was completed, the mixture was cooled to room temperature. 100 mL of purified water and 500 mL of dichloromethane were added to the reaction solution and stirred thoroughly. The mixture was separated, and the organic phase was dried over anhydrous sodium sulfate. The solution was filtered. The filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 40 / 1). The chromatographic solution was concentrated to dryness to obtain an amorphous product with a purity of 97.32%.

[0169] The amorphous XRPD spectrum is shown in Figure 1.

[0170] Example 2 Biological Test Evaluation:

[0171] Test Example 1. ZR-75-1 Cell Proliferation Experiment

[0172] 1. ZR-75-1 cells were seeded in 96-well plates and cultured overnight at 37°C in a 5% CO2 incubator.

[0173] 2. Dilute the compound with culture medium and add it to a 96-well plate. Incubate the plate and continue culturing. Re-digest the cells every 5 days, count the cells, and plate them. Once the cells have completely adhered to the plate, add the compound again and incubate for a total of 10 days.

[0174] 3. After the culture is complete, add 60 μL of CellTiter-Glo reagent (Promega, G7573) to each well, shake for 2 minutes in the dark to lyse the cells, and continue to incubate at room temperature in the dark for another 30 minutes.

[0175] 4. Use BMG PHERAstar FSX to read the optical signal value and perform IC calculations using the nonlinear fitting-four-parameter formula in the software. 50 Fitting.

[0176] The test results are shown in Table 11 below:

[0177] Table 11. Proliferation inhibition data of ZR-75-1

[0178] Conclusion: The compound of formula (I) described in this application has good cell proliferation inhibitory activity.

[0179] Test Example 2. Histone H3 Acetylation Experiment

[0180] 1. ZR-75-1 cells were seeded in 384-well plates and cultured overnight at 37°C in a 5% CO2 incubator.

[0181] 2. Add the diluted compound to a 384-well plate and incubate for another 72 hours.

[0182] 3. Discard the supernatant, add paraformaldehyde, and incubate at room temperature to fix the cells.

[0183] 4. After washing twice with PBS, add a membrane permeabilizing agent and incubate for 10 minutes to allow cell membrane permeabilization.

[0184] 5. After washing twice with PBS, add blocking buffer and incubate at room temperature for 1 hour to perform blocking treatment.

[0185] 6. Discard the blocking solution, add the primary antibody (recombinant Anti-Histone H3 (acetyl K23) antibody and Anti-Histone H4 antibody) diluted in the blocking solution, and incubate overnight at 4°C.

[0186] 7. After washing three times with PBST (0.05% Tween-20 in PBS), add the fluorescent secondary antibody diluted with blocking buffer (IRDye 680RD Goat anti-MOUSE, Licor, 926-68070; IRDye 800CW Goat anti-Rabbit, Licor, 926-32211), incubate at room temperature in the dark for 1 hour, and then wash three times with PBST.

[0187] 8. Scanning was performed using an Odyssey CLx microscope, and the fluorescence signal values ​​of each well were quantized. IC50 analysis was then performed using a nonlinear fitting-four-parameter formula in GraphPad Prism 8 software. 50 Fitting.

[0188] The test results are shown in Table 12 below:

[0189] Table 12 ZR-75-1H3K23 acetylation inhibition data

[0190] Conclusion: The compound of formula (I) of this application has significant cell-targeting activity.

[0191] Test Example 3. In vivo pharmacodynamic study of test drug compound (I) and Compound A in a BALB / c nude mouse model of subcutaneous transplantation of human breast cancer ZR-75-1 cells.

[0192] 1. Experimental Objective

[0193] The purpose of this experiment was to evaluate the in vivo efficacy of the test drug compound (I) and Compound A in a BALB / c nude mouse model of subcutaneous transplanted human breast cancer ZR-75-1 cells.

[0194] 2. Laboratory animals

[0195] Species: Mouse

[0196] Strain: BALB / c nude mice

[0197] Age and weight: 6-8 weeks old, weight 19-26 grams

[0198] Sex: Female

[0199] Supplier: Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0200] Compound of Formula I:

[0201] Compound A: Prepared according to patent CN114364672A.

[0202] 3. Experimental methods and procedures

[0203] 3.1 Cell Culture

[0204] Human breast cancer ZR-75-1 (ATCC, CRL-1500) cells were cultured in vitro in adherent form under the following conditions: 1640 medium supplemented with 10% FBS, 100 U / mL penicillin, and 100 μg / mL streptomycin, at 37°C in a 5% CO2 cell culture incubator. Routine passages were performed twice a week. When cell saturation reached 80%-90% and the desired number was achieved, cells were harvested, counted, and seeded.

[0205] 3.2 Estrogen tablet inoculation and urination

[0206] Three days before cell inoculation, 17β-estrogens (0.36 mg) were injected into the left hindquarters of each mouse. One week after inoculation, the animals urinated 2-3 times per week, and if necessary, urinated daily.

[0207] 3.3 Tumor cell inoculation

[0208] Will contain 10×10 6 One 100 μL ZR-75-1 cell was mixed with 100 μL Matrigel (final volume 200 μL) and subcutaneously inoculated into the right posterior back (upper side) of each mouse. On day 18 post-inoculation, the average tumor volume reached 138 mmHg. 3 Dosing was initiated in groups at a time, selecting tumors ranging from 100 to 212 mm in volume. 3 Mice were enrolled and randomly divided into groups of 7 mice each, based on their body weight and tumor volume. The day of grouping was designated PG-D0, and all groups started medication from PG-D0. The experimental grouping and medication regimen are shown in Table 13.

[0209] 3.4 Preparation of the test substance

[0210] Table 13. Preparation methods of test substances Note: The medication should be gently and thoroughly mixed before administration.

[0211] 3.5 Tumor Measurement and Laboratory Indicators

[0212] The experimental endpoint is to examine whether tumor growth is inhibited, delayed, or cured. Tumor diameter is measured three times a week using calipers. The formula for calculating tumor volume is: V = 0.5a × b 2 , where a and b represent the long and short diameters of the tumor, respectively.

[0213] The antitumor efficacy of the compound was evaluated using TGI (%) or relative tumor proliferation rate (T / C) (%). TGI (%) reflects the tumor growth inhibition rate. The calculation of TGI (%) is as follows: TGI (%) = [1 - (mean tumor volume at the end of treatment in a certain treatment group - mean tumor volume at the beginning of treatment in that treatment group) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the beginning of treatment in the solvent control group)] × 100%.

[0214] Relative tumor proliferation rate T / C (%): The calculation formula is as follows: T / C% = T RTV / C RTV ×100% (T) RTV Treatment group RTV; C RTV The negative control group (RTV) was used to calculate the relative tumor volume (RTV) based on tumor measurements. The formula was RTV = V. t / V0, where V0 is the tumor volume of each mouse measured at the time of group administration (i.e., d0), V t V0 represents the tumor volume of each mouse at a given measurement. The mean value for each group is then calculated. V0 and V1... t Tumor volume data were obtained from the same mouse.

[0215] After the experiment, tumor weight will be measured and T will be calculated. weight / C weight Percentage, T weight and C weight The values ​​represent the tumor weight in the drug administration group and the solvent control group, respectively.

[0216] 3. Experimental Results

[0217] 3.1 Weight Changes

[0218] The effects of the test drug compound (I) and Compound A on the body weight of female BALB / c nude mice with subcutaneous transplantation of human breast cancer ZR-75-1 cells are shown in Figure 17.

[0219] 3.2 Tumor growth curve

[0220] Figure 18 shows the tumor growth curves of female BALB / c nude mice with subcutaneous transplantation of ZR-75-1 cells after treatment with the test drug compound (I) and Compound A.

[0221] 3.3 Evaluation Indicators for Antitumor Drug Efficacy

[0222] Table 8 evaluates the antitumor efficacy of compound (Ⅰ) and Compound A in the ZR-75-1 xenograft model.

[0223] Table 14 Summary of data calculated based on tumor volume on day 21 after drug administration Note: T / C represents the relative tumor proliferation rate, T / C (%) = T RTV / C RTV ×100%; TGI is the tumor growth inhibition rate, TGI(%) = [1 - (average tumor volume of a certain treatment group - average tumor volume of the treatment group at the beginning of treatment) / (average tumor volume of the solvent control group - average tumor volume of the solvent control group at the beginning of treatment)] ×100%;

[0224] 4. Conclusion

[0225] In summary, in this experiment, the human breast cancer ZR-75-1 model tumor-bearing mice were resistant to the test drugs compound 2, compound 5-P2, and compound 3, and all the test drugs exhibited significant antitumor effects. Compounds of formula (I) showed more significant antitumor effects compared to Compound A.

[0226] Example 3 Preparation of crystal form A

[0227] 14 g of formula (I) was added to 56 mL of DMSO (4 mL / g, volume ratio of formula (I) compound), heated to 80-85 °C, and stirred until the solid dissolved. The mixture was filtered while hot, and the filter cake was washed with 14 mL of DMSO (1 mL / g, volume ratio of formula (I) compound). The system was kept at 55-60 °C, and 280 mL of ethanol (20 mL / g, volume ratio of formula (I) compound) was added, and the mixture was stirred to crystallize for 60 ± 10 min. The system was cooled to 0 ± 5 °C, and the mixture was stirred to crystallize for 2 ± 1 h. The mixture was filtered, and the filter cake was washed with 40 mL of ethanol (2.86 mL / g, volume ratio of formula (I) compound). The wet product was vacuum dried at 70 ± 5 °C for 6 h to obtain 12.9 g of product, with a yield of 92.1%. XRPD and TGA-DSC analysis showed that the solid was crystal form A. The single crystal structure is shown in Figure 2, and the XRPD spectrum is shown in Figure 3. The TGA-DSC spectrum is shown in Figure 4.

[0228] Example 4 Preparation of crystal form A

[0229] Add 2g of formula (I) to 10mL of dichloromethane (5mL / g volume ratio of formula (I)), heat to 30-35℃, and stir until the solid dissolves; filter while hot, and wash the filter cake with 10mL of dichloromethane (5mL / g volume ratio of formula (I)); maintain the system temperature at 30-35℃, add 200mL of ethanol (100mL / g volume ratio of formula (I)); stir to crystallize for 60±10min. Cool the system to 0±5℃ and stir to crystallize for 2±1h. Filter by suction, wash the filter cake with 6mL of ethanol (3mL / g volume ratio of formula (I)), and dry the wet product under vacuum at 70±5℃ for 6h to obtain 1.72g of product, which is crystal form A. The XRPD spectrum is shown in Figure 3.

[0230] Example 5 Preparation of crystal form A

[0231] 0.5 kg of formula (I) was added to 2 L of DMSO (4 mL / g of formula (I) by volume), heated to 80-85 °C, and stirred until the solid dissolved. The mixture was filtered while hot, and the filter cake was washed with 0.5 L of DMSO (1 mL / g of formula (I) by volume). The system was kept at 55-60 °C, and 10 L of ethanol (20 mL / g of formula (I) by volume) was added, and the mixture was stirred to crystallize for 60 ± 10 min. The system was then cooled to 0 ± 5 °C, and the mixture was stirred to crystallize for 2 ± 1 h. The mixture was filtered, and the filter cake was washed with 1.5 L of ethanol (3 mL / g of formula (I) by volume). The wet product was then vacuum dried at 70 ± 5 °C for 6 h to obtain 0.465 kg of product, which was crystal form A. The XRPD spectrum is shown in Figure 3.

[0232] Example 6 Preparation of crystal form A

[0233] Add 1 g of formula (I) to 4 mL of DMSO (4 mL / g, volume ratio of formula (I) compound), heat to 80-85 °C, and stir until the solid dissolves. Filter while hot, and wash the filter cake with 1 mL of DMSO (1 mL / g, volume ratio of formula (I) compound). Maintain the system temperature at 55-60 °C, add 10 mL of ethyl acetate (10 mL / g, volume ratio of formula (I) compound), and stir to crystallize for 60 ± 10 min. Cool the system to 0 ± 5 °C and stir to crystallize for 2 ± 1 h. Filter by suction, wash the filter cake with 3 mL of ethyl acetate (3 mL / g, volume ratio of formula (I) compound), and dry the wet product under vacuum at 70 ± 5 °C for 6 h to obtain 0.9 g of product, which is crystal form A. The XRPD spectrum is shown in Figure 3.

[0234] Example 7 Preparation of crystal form A

[0235] Add 1 g of formula (I) to 4 ml of DMSO (4 mL / g, volume ratio of formula (I) compound), heat to 80-85 °C, and stir until the solid dissolves. Filter while hot, and wash the filter cake with 1 ml of DMSO (1 mL / g, volume ratio of formula (I) compound). Control the system temperature at 55-60 °C, add 20 ml of purified water (20 mL / g, volume ratio of formula (I) compound), and stir to crystallize for 60 ± 10 min. Cool the system to 0 ± 5 °C and stir to crystallize for 2 ± 1 h. Filter by suction, wash the filter cake with 3 mL of purified water (3 mL / g, volume ratio of formula (I) compound), and dry the wet product under vacuum at 70 ± 5 °C for 6 h to obtain 0.85 g of product, crystal form A. This solid is crystal form A. The XRPD spectrum is shown in Figure 3.

[0236] Example 8 Preparation of crystal form A

[0237] Add 1 g of formula (I) to 4 ml of DMSO (4 mL / g, volume ratio of formula (I) compound), heat to 80-85 °C, and stir until the solid dissolves. Filter while hot, and wash the filter cake with 1 ml of DMSO (1 mL / g, volume ratio of formula (I) compound). Control the system temperature at 55-60 °C, add 20 ml of acetone (20 mL / g, volume ratio of formula (I) compound), and stir to crystallize for 60 ± 10 min. Cool the system to 0 ± 5 °C and stir to crystallize for 2 ± 1 h. Filter by suction, wash the filter cake with 3 mL of acetone (3 mL / g, volume ratio of formula (I) compound), and dry the wet product under vacuum at 70 ± 5 °C for 6 h to obtain 0.88 g of product, which is crystal form A. The XRPD spectrum is shown in Figure 3.

[0238] Example 9 Preparation of crystal form A

[0239] Add 1 g of formula (I) to 4 mL of DMSO (4 mL / g, volume ratio of formula (I) compound), heat to 80-85 °C, and stir until the solid dissolves. Filter while hot, and wash the filter cake with 1 mL of DMSO (1 mL / g, volume ratio of formula (I) compound). Maintain the system temperature at 55-60 °C, add 20 mL of tetrahydrofuran (20 mL / g, volume ratio of formula (I) compound), and stir to crystallize for 60 ± 10 min. Cool the system to 0 ± 5 °C and stir to crystallize for 2 ± 1 h. Filter by suction, wash the filter cake with 3 mL of tetrahydrofuran (3 mL / g, volume ratio of formula (I) compound), and dry the wet product under vacuum at 70 ± 5 °C for 6 h to obtain 0.87 g of product, which is crystal form A. The XRPD spectrum is shown in Figure 3.

[0240] Example 10 Hydrochloride Crystal I

[0241] 0.5 mL of ethyl acetate (volume-to-mass ratio of compound (I) to 0.0400 g of free alkali crystal form A) was added. 7.5 μL of hydrochloric acid (volume-to-mass ratio of compound (I) to 0.1875 mL / g) was added with stirring at room temperature, and the mixture was stirred for 2 days at room temperature. XRD characterization revealed hydrochloride crystal I, as shown in Figure 9, and its TGA-DSC spectrum is shown in Figure 10. 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 9.70 (s, 1H), 7.894-7.890 (d, J = 1.6Hz, 1H), 7.510-7.507 (d, J = 1. 2Hz,1H),7.46-7.43(d,J=8.8Hz,1H),6.83(s,1H),6.77(s,1H),6.73-6.70(d,J=8.4Hz,1H),6.32- 6.31(t,J=2.0Hz,1H),5.45(s,2H),4.29-4.26(d,J=10.4Hz,1H),3.87(s,3H),3.72(s,3H),3.61-3 .58(d,J=10.8Hz,2H),2.04-1.99(m,1H),1.80-1.72(m,1H),0.98-0.94(m,1H),0.67-0.63(m,1H). MS(ESI)M / Z:483.2[M-36.5] + .

[0242] Example 11 Methanesulfonate Crystals I

[0243] 0.5 mL of ethyl acetate (volume ratio of 12.6 mL / g to compound (I)) was added to 0.0397 g of free base crystal form A. 2.9 μL of methanesulfonic acid (volume ratio of 0.073 mL / g to compound (I)) was added with stirring at room temperature, and the mixture was stirred for 2 days. XRD analysis revealed methanesulfonate crystal I, as shown in Figure 11, and its TGA-DSC spectrum is shown in Figure 12. NMR results showed the presence of 1.6 equivalents of methanesulfonic acid. 1H-NMR (400MHz, DMSO-d6): δ (ppm) 9.70 (s, 1H), 7.893-7.887 (d, J = 1.6Hz, 1H), 7.510-7.507 (d, J = 1. 2Hz,1H),7.46-7.43(d,J=8.8Hz,1H),6.83(s,1H),6.77(s,1H),6.73-6.70(d,J=8.4Hz,1H),6.32- 6.31(t,J=2.0Hz,1H),5.45(s,2H),4.29-4.26(d,J=10.4Hz,1H),3.87(s,3H),3.72(s,3H),3.61-3 .58(d,J=10.8Hz,2H),2.04-1.99(m,1H),1.80-1.72(m,1H),0.98-0.94(m,1H),0.67-0.63(m,1H). MS(ESI)M / Z:483.1[M-96] + .

[0244] Example 12 p-Toluenesulfonate crystals I

[0245] 0.5 mL of ethyl acetate (volume-to-mass ratio of 12.63 mL / g to compound (I)) was added to 0.0396 g of free base crystal form A. 15.4 mg of p-toluenesulfonic acid (0.39 times the mass of free base crystal form A) was added with stirring at room temperature, and the mixture was stirred for 2 days. XRD analysis revealed p-toluenesulfonate crystal form I, as shown in Figure 5, and its TGA-DSC spectrum is shown in Figure 6. NMR results showed the presence of 1.1 equivalents of p-toluenesulfonic acid. 1H-NMR (400MHz, DMSO-d6): δ (ppm) 9.70 (s, 1H), 7.894-7.890 (d, J = 1.6Hz, 1H), 7.512-7.509 (d, J = 1.2Hz, 1H), 7.49-7 .47(d,J=8.0Hz,2H),7.46-7.43(d,J=8.8Hz,1H),7.14-7.12(d,J=8.0Hz,2H),6.83(s,1H),6.77(s,1H),6.73-6.71( d,J=8.4Hz,1H),6.32-6.31(t,J=2.0Hz,1H),5.45(s,2H),4.29-4.26(d,J=10.4Hz,1H),3.88(s,3H),3.72(s,3H),3. 61-3.58(d,J=10.8Hz,2H),2.29(s,3H),2.03-1.99(m,1H),1.80-1.72(m,1H),0.97-0.94(m,1H),0.67-0.63(m,1H). MS(ESI)M / Z:483.2[M-172] + .

[0246] Example 13 Potassium Salt Crystal I

[0247] 1 mL of methanol (volume-to-mass ratio of 25 mL / g to compound (I)) was added to 0.0399 g of free alkali crystal form A. 5.0 mg of potassium hydroxide (0.125 times the mass of free alkali crystal form A) was added with stirring at room temperature for 2 days. XRD characterization revealed potassium salt crystal form I, as shown in Figure 13, and its TGA-DSC spectrum is shown in Figure 14. 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 7.864-7.858 (d, J = 2.4Hz, 1H), 7.492-7.488 (d, J = 1.2Hz, 1H), 7.15 (brs, 1H), 6.55 (brs, 3H), 6.3 0-6.29(t,J=2.0Hz,1H),5.38(s,2H),3.96(brs,1H),3.84(s,3H),3.63(s,3H),1.91(brs,1H),1.63(brs,1H),0.86-0.76(m,2H). MS(ESI)M / Z:483.1[M-39] + .

[0248] Example 7 Potassium Salt Crystal II

[0249] 0.5 mL of butanone (volume-to-mass ratio of 12.5 mL / g to compound (I)) was added to 0.0400 g of free alkali crystal form A. 5.4 mg of potassium hydroxide (0.135 times the mass of free alkali crystal form A) was added with stirring at room temperature, and the mixture was stirred for 2 days at room temperature. XRD characterization revealed potassium salt crystal form II, as shown in Figure 7, and its TGA-DSC spectrum is shown in Figure 8. 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 7.852-7.847 (d, J = 2.0Hz, 1H), 7.482-7.478 (d, J = 1. 6Hz,1H),7.16-7.14(d,J=8.8Hz,1H),6.56-6.51(m,3H),6.29-6.28(t,J=2.0Hz,1H),5 .37(s,2H),4.04-4.02(d,J=10.8Hz,1H),3.83(s,3H),3.63(s,3H),3.33-3.31(d,J=11 .2Hz,2H),1.91-1.87(m,1H),1.67-1.56(m,1H),0.88-0.84(m,1H),0.79-0.77(m,1H). MS(ESI)M / Z:483.1[M-39] + .

[0250] Example 8 Choline Salt Crystals I

[0251] 0.5 mL of tetrahydrofuran (volume-to-mass ratio of 12.4 mL / g to compound (I)) was added to 0.0402 g of crystal form A. Then, 19.5 μL of choline aqueous solution (40% by mass) (0.211 times the mass of free base crystal form A) was added with stirring at room temperature. The mixture was stirred at room temperature for 2 days. XRD characterization revealed choline salt crystal I, as shown in Figure 15, and its TGA-DSC spectrum is shown in Figure 16. NMR results showed the presence of 0.94 equivalents of choline. 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 7.87-7.86 (d, J = 2.0Hz, 1H), 7.493-7.490 ( d,J=1.2Hz,1H),7.15-7.13(brs,1H),6.54-6.52(brs,3H),6.30-6.29(t,J= 2.0Hz,1H),5.38(s,2H),3.89-3.79(m,6H),3.59(s,3H),3.53-3.48(m,1H), 1.93-1.88(m,1H),1.67-1.53(m,1H),0.87-0.82(m,1H),0.75-0.71(m,1H). MS(ESI)M / Z:483.1[M-104] +.

[0252] Example 10 Hygroscopicity Test:

[0253] Referring to the "Guiding Principles for Hygroscopicity Testing of Drugs" in the Chinese Pharmacopoeia, the water adsorption / desorption data of crystal form A were tested.

[0254] Figure 19 shows the DVS curve of crystal form A. The DVS results show that crystal form A has a moisture absorption weight gain of 0.68% at 80% RH, indicating that the crystal form is slightly hygroscopic. The XRPD of the remaining solid after the DVS experiment showed that the crystal form did not change.

[0255] Example 11 Stability test:

[0256] Following the guidelines for stability testing of active pharmaceutical ingredients and formulations in the Chinese Pharmacopoeia, the stability of crystal form A under accelerated conditions was investigated. Purity was determined by HPLC on days 0 and 10, and crystal form was determined by XRPD.

[0257] Table 15. Experimental results of crystal form A stability

[0258] The experimental results are shown in Table 15. Crystal form A has stable physical properties under high temperature and high humidity conditions, its crystal form has not changed, and its chemical purity has not decreased significantly.

[0259] Table 16 Results of Potassium Salt Crystal II Stability Tests

[0260] The experimental results are shown in Table 16. Potassium salt crystal II has stable physical properties under high temperature and high humidity conditions, its crystal form has not changed, and its chemical purity has not decreased significantly.

Claims

1. Crystal form A of compound of formula (Ⅰ): Its features are, Using Cu-Kα radiation, the X-ray powder diffraction pattern of crystal form A has characteristic peaks at 2θ values ​​of 12.34°, 15.63°, 18.64°, 20.09°, 21.62°, 22.94°, and 25.38°, with a 2θ error range of ±0.2°.

2. The crystal form A as described in claim 1, characterized in that, Using Cu-Kα radiation, the X-ray powder diffraction pattern shows characteristic peaks at 2θ values ​​of 9.51°, 12.34°, 15.63°, 16.77°, 18.64°, 20.09°, 21.42°, 21.62°, 22.61°, 22.94°, 24.28°, 25.38°, 25.71°, 26.26°, 26.72°, and 32.28°, with a 2θ error range of ±0.2°.

3. Crystal form A as described in claim 1, characterized in that, Using Cu-Kα radiation, the X-ray powder diffraction pattern shows characteristic peaks at 2θ values ​​of 9.51°, 11.19°, 11.79°, 12.34°, 12.71°, 13.22°, 15.18°, 15.63°, 16.77°, 18.64°, 19.20°, 20.09°, 21.42°, 21.62°, 22.61°, 22.94°, 24.28°, 25.38°, 25.71°, 26.26°, 26.72°, 28.9°, 30.10°, 31.44°, 32.28°, and 37.68°, with a 2θ error range of ±0.2°.

4. Crystal form A as described in claim 1, characterized in that, The X-ray powder diffraction pattern peak analysis data of the crystal form are shown in Table 1. More preferably, its X-ray powder diffraction pattern is shown in Figure 3, with a 2θ error range of ±0.2°.

5. Crystal form A as described in claim 1, characterized in that, The TGA-DSC spectrum of the crystal form shows a weight loss of 29.59% at 35.46±3 to 351.51±3 °C, preferably at 35.46±2 to 351.51±2 °C, for example at 35.46 to 351.51.51 °C.

6. Crystal form A as described in claim 1, characterized in that, The TGA-DSC spectrum of the crystal form has an endothermic peak at 197.75±5℃ and an exothermic peak at 268.77±5℃. Preferably, it has an endothermic peak at 197.75±2℃ and an exothermic peak at 268.77±2℃. For example, it has an endothermic peak at 197.75℃ and an exothermic peak at 268.77℃.

7. Crystal form A as described in claim 1, characterized in that, The TGA-DSC spectrum of the crystal form is shown in Figure 4.

8. The p-toluenesulfonate crystal form I of the compound of formula (I) or the potassium salt crystal form II of the compound of formula (I), characterized in that: In p-toluenesulfonate crystal form I, the molar ratio of free base to p-toluenesulfonic acid is 1:1; More preferably, using Cu-Kα radiation, the X-ray powder diffraction pattern of crystal form I has characteristic peaks at 2θ values ​​of 11.20, 12.80, 15.75, 16.12, 20.17, and 24.32, with a 2θ error range of ±0.2°; More preferably, using Cu-Kα radiation, the X-ray powder diffraction pattern shows characteristic peaks at 2θ values ​​of 8.04, 11.20, 12.40, 12.80, 13.72, 15.75, 16.12, 20.17, 20.92, 21.93, 24.32, and 25.15, with a 2θ error range of ±0.2°. More preferably, using Cu-Kα radiation, the X-ray powder diffraction pattern shows characteristic peaks at 2θ values ​​of 8.04, 11.20, 11.94, 12.40, 12.80, 13.72, 15.30, 15.75, 16.12, 18.42, 18.74, 19.42, 20.17, 20.92, 21.93, 22.32, 22.74, 23.51, 24.32, and 25.15, with a 2θ error range of ±0.2°. More preferably, the X-ray powder diffraction pattern peak analysis data of the crystal form are shown in Table 2; More preferably, its X-ray powder diffraction pattern is shown in Figure 5, with a 2θ error range of ±0.2°; More preferably, the TGA-DSC spectrum of the crystal form shows a weight loss of 33.29% at 35.61±3 to 351.92±3℃, more preferably, a weight loss of 33.29% at 35.61±2 to 351.92±2℃, for example, a weight loss of 33.29% at 35.61 to 351.92℃; More preferably, the TGA-DSC spectrum of the crystal form has an endothermic peak at 145.85±5℃, more preferably, an endothermic peak at 145.85±2℃, for example, an endothermic peak at 145.85℃; More preferably, the TGA-DSC spectrum of the crystal form is shown in Figure 6; In the potassium salt crystal form II of compound (I), using Cu-Kα radiation, the X-ray powder diffraction pattern of crystal form II has characteristic peaks at 2θ values ​​of 9.85, 15.32, 19.44, 20.50, 20.86, and 22.81, with a 2θ error range of ±0.2°; More preferably, using Cu-Kα radiation, the X-ray powder diffraction pattern shows characteristic peaks at 2θ values ​​of 8.86, 9.85, 10.93, 14.70, 15.32, 16.54, 19.44, 20.50, 20.86, 22.81, 25.67, and 27.06, with a 2θ error range of ±0.2°. More preferably, using Cu-Kα radiation, the X-ray powder diffraction pattern shows characteristic peaks at 2θ values ​​of 8.86, 9.85, 10.93, 12.25, 12.90, 13.88, 14.70, 15.32, 16.54, 17.85, 19.44, 20.50, 20.86, 21.77, 22.81, 25.67, 27.06, 27.63, 31.37, 32.77, and 34.21, with a 2θ error range of ±0.2°. More preferably, the X-ray powder diffraction pattern analysis data of the crystal form are shown in Table 3; More preferably, its X-ray powder diffraction pattern is shown in Figure 7, with a 2θ error range of ±0.2°; More preferably, the TGA-DSC spectrum of the crystal form shows a weight loss of 21.97% at 35.56±3 to 352.05±3℃, more preferably, a weight loss of 21.97% at 35.56±2 to 352.05±2℃, for example, a weight loss of 21.97% at 35.56 to 352.05℃; More preferably, the TGA-DSC spectrum of the crystal form has an exothermic peak at 260.64℃±5℃, more preferably an exothermic peak at 260.64℃±2℃, for example, an exothermic peak at 260.64℃; More preferably, the TGA-DSC spectrum of the crystal form is shown in Figure 8.

9. A pharmaceutical composition comprising the crystal form according to any one of claims 1-8 and a pharmaceutically acceptable carrier.

10. Use of the crystal form of any one of claims 1-8 or the pharmaceutical composition of claim 9 in the preparation of a medicament for treating KAT6-mediated cancer.

11. Use of the crystal form of any one of claims 1-8 or the pharmaceutical composition of claim 9 for the treatment of KAT6-mediated cancer.

12. The use as described in claim 10 or 11, wherein the cancer includes breast cancer, prostate cancer, and lung cancer.

13. The method for preparing crystal form A according to any one of claims 1-7, characterized in that, Includes the following steps: (a) Dissolve the compound of formula (Ⅰ) in a first solvent, heat, stir, filter, and wash; (b) At a certain temperature, add the second solvent to the solution in step (a) and stir for a certain time; (c) Cool down, stir, stir for a certain period of time, filter, wash, collect, and dry the solid.

14. The preparation method according to claim 13, wherein the first solvent is selected from one or a mixture of two of dichloromethane and dimethyl sulfoxide; preferably dimethyl sulfoxide.

15. The preparation method as described in claim 13, wherein the second solvent is selected from one or more mixtures of acetone, butanone, methyl isobutyl ketone, methanol, ethanol, isopropanol, ethyl acetate, isopropyl acetate, tetrahydrofuran, 1,4-dioxane, methyl tert-butyl ether, water, and acetonitrile; preferably ethanol.

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