A crystalline form of a 5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine compound and a method of preparing the same

Abstract

A crystal form of a 5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine compound and a preparation method thereof, and application of the crystal form in preparation of a drug for treating related diseases are disclosed. Specifically disclosed are a crystal form of a compound of formula (I) and a preparation method thereof.

Description

Technical Field

[0001] This invention relates to a crystal form of a 5,6-dihydrobenzimido[f]imidazole[1,2-d][1,4]oxaconitine compound and a method for preparing the same, and also to the application of said crystal form in the preparation of medicaments for treating related diseases. Specifically, it relates to the crystal form of compound (I) and a method for preparing the same. Background Technology

[0002] Phosphatidylinositol-3-kinase (PI3K) is a lipokinase composed of the regulatory subunit p85 or p101 and the catalytic subunit p110 (which is further divided into four isoforms: p110a, p110b, p110g, and p110d). It plays a crucial role in cell proliferation, survival, and metabolism by catalyzing the phosphorylation of the 3'-OH group of phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidylinositol 3,4,5-trisphosphate (PIP3), thereby activating downstream Akt and other kinases. In tumor cells, PI3K overexpression leads to rapid proliferation and growth of tumor cells.

[0003] There are four subtypes of PI3K, among which PI3Kα is widely distributed in the body. Aberrant activation of PI3Kα has also been found in various solid tumors. Mutations in the PIK3CA gene also exist in different solid tumors, contributing to tumor development and progression. In normal physiological function, PI3Kα primarily regulates insulin and related glucose regulation pathways. Therefore, inhibition of wild-type PI3Kα has been clinically verified to cause side effects such as hyperglycemia. Thus, inhibitors targeting mutant PI3Kα play a crucial role in ensuring clinical safety.

[0004] GDC-0077 is a highly selective PI3Kα inhibitor developed by Roche. It also has the function of degrading mutant PI3Kα protein, which brings new hope for the clinical development of PI3K inhibitors with higher safety.

[0005] Summary of the Invention

[0006] This invention provides crystal form A of the compound of formula (I), characterized by its X-ray powder diffraction pattern exhibiting characteristic diffraction peaks at the following 2θ angles: 5.25±0.20°, 8.74±0.20°, 10.46±0.20°.

[0007]

[0008] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 5.25±0.20°, 7.78±0.20°, 8.74±0.20°, 10.46±0.20°, and 18.48±0.20°.

[0009] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 5.25±0.20°, 7.78±0.20°, 8.74±0.20°, 10.46±0.20°, 14.99±0.20°, 16.25±0.20°, 17.50±0.20°, and 18.48±0.20°.

[0010] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 5.25±0.20°, 8.74±0.20°, and / or 10.46±0.20°, and / or 6.65±0.20°, and / or 7.78±0.20°, and / or 12.60±0.20°, and / or 13.26±0.20°, and / or 13.87±0.20°, and / or 14.44±0.20°, and / or 14.99±0.20°, and / or 15.53±0.20°, and / or 16.25±0.20°, and / or 17.50±0.20°, and / or 18.48±0.20°, and / or 19.01±0.20°. , and / or 19.78±0.20°, and / or 20.55±0.20°, and / or 21.01±0.20°, and / or 21.33±0.20°, and / or 23.72±0.20°, and / or 23.96±0.20°, and / or 24.46±0.20°, and / or 24.79±0.20°, and / or 25.0 2±0.20°, and / or 25.69±0.20°, and / or 26.00±0.20°, and / or 26.29±0.20°, and / or 26.72±0.20°, and / or 27.34±0.20°, and / or 27.58±0.20°, and / or 28.17±0.20°, and / or 28.57±0.20°.

[0011] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 5.2511°, 6.6532°, 7.7817°, 8.7418°, 10.4626°, 12.5958°, 13.2601°, 13.8718°, 14.4446°, 14.9927°, 15.5346°, 16.2492°, 17.5047°, and 18.4752°. , 19.0069°, 19.7791°, 20.5468°, 21.0093°, 21.3325°, 23.7228°, 23.9609°, 24.4579°, 24.7921°, 25.0236°, 25.6910°, 26.0008°, 26.2926°, 26.7249°, 27.3381°, 27.5777°, 28.1708°, 28.5699°.

[0012] In some embodiments of the present invention, the XRPD pattern of the above-mentioned crystal form A is essentially as follows: Figure 1 As shown.

[0013] The present invention also provides the above-mentioned crystal form A, whose XRPD pattern is basically as follows: Figure 1 As shown.

[0014] In some embodiments of the present invention, the XRPD spectra analysis data of the above-mentioned A-type crystal form are shown in Table 1:

[0015] Table 1. XRPD spectrum analysis data of compound A (I)

[0016]

[0017]

[0018]

[0019] In some embodiments of the present invention, the differential scanning calorimetry curves of the above-mentioned A crystal form have an endothermic peak starting point at 90.5±3.0℃ and 181.7±3.0℃, respectively.

[0020] In some embodiments of the present invention, the DSC spectrum of the above-mentioned A-type crystal is as follows: Figure 2 As shown.

[0021] In some embodiments of the present invention, the DSC spectrum of the above-mentioned A-type crystal is essentially as follows: Figure 2 As shown.

[0022] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned A crystal form shows a weight loss of 2.55% at 110.0±3.0℃.

[0023] In some embodiments of the present invention, the TGA pattern of the above-mentioned A crystal form is as follows: Figure 3 As shown.

[0024] In some embodiments of the present invention, the TGA pattern of the above-mentioned A-type crystal is essentially as follows: Figure 3 As shown.

[0025] In some embodiments of the present invention, the above-mentioned A crystal form has a moisture absorption weight gain of 1.093% at 25°C and 80% RH.

[0026] In some embodiments of the present invention, the DVS pattern of the above-mentioned A crystal form is as follows: Figure 7 As shown.

[0027] In some embodiments of the present invention, the DVS pattern of the above-mentioned A crystal form is essentially as follows: Figure 7 As shown.

[0028] The present invention also provides crystal form B of the compound of formula (I), characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.48±0.20°, 21.84±0.20°, 24.90±0.20°.

[0029]

[0030] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned B crystal form has characteristic diffraction peaks at the following 2θ angles: 5.48±0.20°, 9.38±0.20°, 13.25±0.20°, 19.42±0.20°, 21.84±0.20°, 24.90±0.20°.

[0031] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned B crystal form has characteristic diffraction peaks at the following 2θ angles: 5.4752°, 9.3771°, 10.9011°, 13.2511°, 16.3529°, 18.9134°, 19.4248°, 21.8375°, 24.8988°, 26.6143°, and 27.6851°.

[0032] In some embodiments of the present invention, the XRPD pattern of the above-mentioned B crystal form is essentially as follows: Figure 4 As shown.

[0033] The present invention also provides the above-mentioned B crystal form, whose XRPD pattern is basically as follows: Figure 4 As shown.

[0034] In some embodiments of the present invention, the XRPD spectra analysis data of the above-mentioned B crystal form are shown in Table 2:

[0035] Table 2 shows the XRPD spectrum analysis data of compound B (I)

[0036]

[0037]

[0038] In some embodiments of the present invention, the differential scanning calorimetry curve of the B crystal form has an endothermic peak starting point at 83.5±3.0℃.

[0039] In some embodiments of the present invention, the DSC pattern of the above-mentioned B crystal form is as follows: Figure 5 As shown.

[0040] In some embodiments of the present invention, the DSC pattern of the above-mentioned B crystal form is essentially as follows: Figure 5 As shown.

[0041] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned B crystal form shows a weight loss of 6.73% at 135.0±3.0℃.

[0042] In some embodiments of the present invention, the TGA pattern of the above-mentioned B crystal form is as follows: Figure 6 As shown.

[0043] In some embodiments of the present invention, the TGA pattern of the B crystal form described above is essentially as follows: Figure 6 As shown.

[0044] In some embodiments of the present invention, the above-mentioned B crystal form has a moisture absorption weight gain of 5.731% at 25°C and 80% RH.

[0045] In some embodiments of the present invention, the DVS pattern of the above-mentioned B crystal form is as follows: Figure 8 As shown.

[0046] In some embodiments of the present invention, the DVS pattern of the above-mentioned B crystal form is essentially as follows: Figure 8 As shown.

[0047] The present invention also provides the application of the above-mentioned crystal form A and crystal form B in the preparation of drugs for treating advanced solid tumors.

[0048] Definitions and Explanations

[0049] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular phrase or term should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When trade names appear herein, they are intended to refer to the corresponding product or its active ingredient.

[0050] The intermediate compounds of the present invention can be prepared by various 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 the present invention.

[0051] The chemical reactions in the specific embodiments of this invention are carried out in a suitable solvent, which must be suitable for the chemical changes of this invention and the reagents and materials required therefor. To obtain the compounds of this invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0052] Unless otherwise stated, in the differential scanning calorimetry curves of the compounds of the present invention, upward indicates exothermic (Exo Up).

[0053] Unless otherwise stated, in powder X-ray diffraction (XRPD) spectroscopy, the position or relative intensity of peaks may vary due to factors such as the measuring instrument, method / conditions, etc. For any given crystal form, the peak position may have errors, with the 2θ value measurement error potentially reaching ±0.2°. Therefore, this error should be taken into account when determining each crystal form, and such errors are within the scope of this application.

[0054] In this invention, a specific XRPD pattern described as "substantially as shown" means that the positions of the diffraction peaks in the XRPD pattern are substantially the same within a range of ±0.20°, 2θ, as observed visually or by means of a selected list of diffraction peaks. Those skilled in the art will understand that the intensity can vary with the sample.

[0055] The structures of the compounds of this invention can be confirmed by conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, that absolute configuration can be confirmed by conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) is used, where the cultured single crystal is used to collect diffraction intensity data using a Bruker D8 venture diffractometer with CuKα radiation as the light source. The scanning method is as follows: After scanning and collecting relevant data, the crystal structure can be further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0056] The present invention will be described in detail below through embodiments, which are not intended to limit the present invention in any way.

[0057] All solvents used in this invention are commercially available and can be used without further purification.

[0058] The solvent used in this invention is commercially available.

[0059] Compounds are named according to conventional naming principles in the field or using Software naming conventions are used; commercially available compounds use supplier catalog names.

[0060] Technical effect

[0061] Compound (I) effectively inhibits PI3Kα kinase activity and also effectively inhibits cell proliferation in PIK3CA-mutated HCC1954 cells; Compound (I) A has a stable crystal form.

[0062] The present invention relates to an X-ray powder diffractometer (XRPD) method.

[0063] Instrument Model: Panalytical Empyrean X-ray Diffractometer

[0064] Test method: Spread an appropriate amount of sample evenly on a single-crystal silicon sample disk and perform XRPD testing using the parameters described below.

[0065] The experimental parameters are shown in Table 3:

[0066] Table 3 XRPD Instrument Parameters

[0067]

[0068] This invention relates to a differential scanning calorimeter (DSC) method.

[0069] Instrument Model: TA Discovery DSC 2500 Differential Scanning Calorimeter

[0070] The experimental parameters are shown in Table 4:

[0071] Table 4 DSC Instrument Parameters

[0072] parameter Setting value Sample tray Aluminum disc, pressure cap Temperature range 25～315℃ Scan rate (°C / min) 10 Protective gas Nitrogen

[0073] The present invention relates to a thermogravimetric analysis (TGA) method.

[0074] Instrument Model: TA Discovery TGA 5500 Thermogravimetric Analyzer

[0075] The experimental parameters are shown in Table 5:

[0076] Table 5 TGA Instrument Parameters

[0077] parameter Setting value Sample tray Aluminum disc, open the lid Temperature range Room temperature ~ 350℃ Scan rate (°C / min) 10 Protective gas Nitrogen

[0078] This invention presents a method for dynamic moisture adsorption analysis (DVS).

[0079] Dynamic moisture adsorption curves were collected using a DVSIntrinsic instrument from SMS (Surface Measurement Systems) in the UK. Relative humidity at 25°C was corrected for the deliquescence points of lithium chloride (LiCl), magnesium nitrate [Mg(NO3)2], and potassium chloride (KCl).

[0080] Table 6 DVS Instrument Parameters

[0081]

[0082] Hygroscopicity evaluation is classified as follows:

[0083] Absorbs sufficient moisture to form a liquid: deliquescent; ΔW% ≥ 15%: highly hygroscopic; 15% > ΔW% ≥ 2%: hygroscopic; 2% > ΔW% ≥ 0.2%: slightly hygroscopic; ΔW% < 0.2%: no or almost no hygroscopicity. ΔW% represents the weight gain due to moisture absorption at 25℃ / 80%RH. Attached Figure Description

[0084] Figure 1 The XRPD spectrum of Cu-Kα radiation for crystal form A of compound (I) is shown.

[0085] Figure 2 The DSC spectrum of compound A of formula (I) is shown.

[0086] Figure 3 The TGA spectrum of compound A of formula (I) is shown.

[0087] Figure 4 The XRPD spectrum of Cu-Kα radiation for the B crystal form of compound (I) is shown.

[0088] Figure 5 The DSC spectrum of compound B of formula (I) is shown.

[0089] Figure 6 The TGA spectrum of compound B of formula (I) is shown.

[0090] Figure 7 The DVS spectrum of compound A of formula (I) is shown.

[0091] Figure 8 The DVS spectrum of compound B of formula (I) is shown.

[0092] Figure 9 The diagram shows the ellipsoid of the three-dimensional structure of compound (I). Detailed Implementation

[0093] To better understand the content of this invention, further explanation will be provided below with reference to specific embodiments. However, the specific implementation methods are not intended to limit the content of this invention.

[0094] Experimental Example 1: Preparation of Compound (I)

[0095]

[0096] Synthesis route:

[0097]

[0098] Step 1: Synthesis of Compounds 1-2

[0099] Add 1-1 (4.0 g, 9.82 mmol) to a pre-dried reaction flask, then add tetrahydrofuran (200 mL), and finally Lawson's reagent (7.94 g, 19.64 mmol). The reaction was carried out at 20 °C with stirring for 12 hours. After the reaction was complete, water was added dropwise to quench the reaction mixture, followed by extraction with dichloromethane (100 mL x 3). The organic phases were combined, washed with 100 mL of saturated sodium chloride solution, and dried over anhydrous sodium sulfate. After filtration, the crude product was directly concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 20:1). The purified compound was added to an aqueous hydrochloric acid solution (6 M, 100 mL), followed by extraction with 300 mL of dichloromethane. The pH of the aqueous phase was adjusted with sodium carbonate until solids precipitated. The aqueous phase was further extracted with 300 mL of dichloromethane, and the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 1-2.

[0100] 1 H NMR (400MHz, CDCl3) δ = 8.08 (d, J = 8.8Hz, 1H), 8.05-7.98 (m, 1H), 7.39-7.25 (m, 1H) ,7.12-7.08(m,1H),6.77-6.44(m,1H),6.35(dd,J=2.4,8.7Hz,1H),6.15(d,J=2.5 Hz,1H),4.85-4.72(m,1H),4.65(dd,J=4.0,9.5Hz,1H),4.48-4.39(m,1H),4.34(d d,J=2.5,5.0Hz,2H),4.25-4.15(m,3H),1.59(d,J=6.5Hz,3H); MS:m / z=424.1[M+1] + .

[0101] Step 2: Synthesis of compound (I)

[0102] Add 1-2 (20.00 mg, 47.23 μmol) to a pre-dried reaction flask, then add ethanol (5 mL), potassium carbonate (32.64 mg, 236.16 μmol), and O-methylhydroxylamine hydrochloride (19.72 mg, 236.16 μmol). Stir the reaction mixture at 80 °C for 12 hours. After the reaction is complete, concentrate the reaction solution under reduced pressure, add 10 mL of water, and then extract with dichloromethane (10 mL * 3). Combine the organic phases. Wash the organic phase with 10 mL of saturated sodium chloride solution and dry with anhydrous sodium sulfate. Concentrate the organic phase under reduced pressure to obtain compound (I).

[0103] 1H NMR (400MHz, CDCl3) δ=8.11-7.98(m,1H),7.19(s,1H),6.78-6.45(m,1H),6.39(ddd,J=2.4,8.8,13.2H z,1H),6.24(dd,J=2.4,16.4Hz,1H),4.86-4.73(m,1H),4.68-4.57(m,3H),4.49-4.39(m,1H),4.33(br d,J=3.3Hz,2H),4.24-4.16(m,2H),3.94-3.83(m,2H),3.79(s,3H),1.45(d,J=6.5Hz,3H); MS:m / z=437.2[M+1] + .

[0104] Example 2: Preparation of crystal form A of compound (I)

[0105] Weigh 100 mg of compound (I), add n-heptane (2 mL, 20 V) to form a suspension, then add methyl tert-butyl ether (0.2 mL, 2 V), and finally add ethanol (0.2 mL, 2 V). Stir at room temperature for 16 hours, filter, and dry the filter cake under reduced pressure to obtain the A crystal form of compound (I).

[0106] Weigh 53 g of compound (I), add n-heptane (1060 mL, 20 v) to form a suspension, then add methyl tert-butyl ether (106 mL, 2 v), and add ethanol (106 mL, 2 v) while stirring. Finally, add water (1060 mL, 20 v) and stir at room temperature (approximately 25 °C) for 116 hours. Add approximately 5 g of crystalline form A of compound (I), and continue stirring for approximately 95 hours. Filter, and dry the filter cake under reduced pressure to obtain crystalline form A of compound (I).

[0107] Example 3: Preparation of crystal form B of compound (I)

[0108] Weigh 100 mg of compound (I), add n-heptane (1 mL, 10 v) to form a suspension, then add methanol (0.1 mL, 1 v), stir at 25 °C for 16 hours, filter, and dry the filter cake under reduced pressure to obtain the B crystal form of compound (I).

[0109] Example 4: Crystal form competition experiment

[0110] After weighing approximately 0.5 g of compound (I), supersaturating it in the following four groups of solvents, the supernatant was used as the solvent for the competitive study of crystal forms A and B. The four groups of mixed solvents are as follows:

[0111] Group 1: Water (2mL)

[0112] Group 2: n-Heptane: tertiary methyl ether: ethanol = 2 mL: 0.2 mL: 0.2 mL

[0113] Group 3: n-Heptane: Tertiary Methyl Ether: 95% Ethanol = 2 mL: 0.2 mL: 0.2 mL

[0114] Group 4: n-Heptane: Tertiary Methyl Ether: 90% Ethanol = 2 mL: 0.2 mL: 0.2 mL

[0115] 1 mL of supernatant was taken from each of the four supersaturated solutions to conduct the following study. Equal masses of crystal form A (50 mg) and crystal form B (50 mg) were added and stirred at 25 °C and 50 °C respectively. After 5 days, solid samples from the solutions were taken for XRPD characterization to investigate the changes in crystal form.

[0116] The results showed that the crystal form of compound A of formula (I) was obtained in each experimental system under the conditions of 25℃ and 50℃.

[0117] Example 5: Solid stability test of crystal form A of compound (I)

[0118] Weigh 20 mg of compound A (I) into a 40 mL glass bottle, seal the bottle opening with aluminum foil, and then poke several small holes with a needle to ensure sufficient contact between the sample and ambient temperature and humidity (for light conditions, simply leave the bottle open). Place the sample under the corresponding influencing factor test conditions (60℃; 25℃ / 92.5%RH; UV + Visible light) and accelerated conditions (40℃ / 75%RH and 60℃ / 75%RH). Take samples for analysis at 5 days, 10 days, 1 month, 2 months, and 3 months.

[0119] Table 7 shows the solid stability test results of compound A of formula (I).

[0120]

[0121] The results show that the crystal form of compound A of formula (I) is stable under high temperature, high humidity, light irradiation and accelerated conditions.

[0122] Example 6: Hygroscopicity study of crystal form A of formula (I)

[0123] Experimental materials:

[0124] SMSDVS intrinsic dynamic moisture adsorption meter

[0125] Experimental methods:

[0126] Take 10-30 mg of compound A (I) and place it in the DVS sample tray for testing.

[0127] Experimental results:

[0128] The DVS spectrum of compound A of formula (I) is as follows: Figure 7 As shown, ΔW = 1.093%.

[0129] Experimental conclusion:

[0130] The hygroscopic weight gain of compound A in formula (I) at 25°C and 80% RH is 1.093%.

[0131] Example 7: Hygroscopicity study of crystal form B of formula (I)

[0132] Experimental materials:

[0133] SMS DVS Intrinsic Dynamic Moisture Adsorption Meter

[0134] Experimental methods:

[0135] Take 10-30 mg of compound B (I) and place it in the DVS sample tray for testing.

[0136] Experimental results:

[0137] The DVS spectrum of compound B of formula (I) is as follows: Figure 8 As shown, ΔW = 5.731%.

[0138] Experimental conclusion:

[0139] The hygroscopic weight gain of compound B of formula (I) at 25°C and 80% RH is 5.731%.

[0140] Example 8: Single-crystal X-ray diffraction analysis of compound (I)

[0141]

[0142] Single crystal cultivation process: 10 mg of compound (I) was added to 2 mL of ethanol and stirred until the sample was completely dissolved, resulting in a colorless clear solution. The sample solution was placed in a 4 mL semi-sealed sample vial and allowed to evaporate slowly at room temperature. After two weeks, colorless needle-like crystals were obtained. The crystals were collected, and diffraction intensity data were collected using a single-crystal X-ray diffractometer (SC-XRD) (D8-VENTURE). The single-crystal data showed that the absolute configuration of compound (I) could be determined, and the molecular formula of the compound was C10. 19 H 22 F2N6O4. The stereoscopic ellipsoid diagram of compound (I) is shown below. Figure 9 The crystal structure data of single crystals of compound (I) are shown in Table 8.

[0143] Table 8 Crystal data of single crystals of compound (I)

[0144]

[0145]

[0146] Experimental Example 1: In vivo efficacy evaluation

[0147] 1. Experimental objective: To evaluate the in vivo efficacy of the test drug in a subcutaneous xenograft tumor model of human breast cancer cells HCC1954.

[0148] 2. Experimental Design:

[0149] (1) Cell culture: Human breast cancer HCC1954 cells were cultured in vitro in a monolayer with 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin added to the appropriate culture medium, and incubated at 37℃ in a 5% CO2 incubator. Routine treatment and passage were performed twice a week. When the cell saturation reached 80%-90% and the required number was achieved, the cells were harvested, counted, and seeded.

[0150] (2) Animals: Balb / c nuds, female, 6–8 weeks old, weighing 18–20 grams. A total of 45 animals are required (including any remaining mice from the grouping). These will be provided by Shanghai Jihui Laboratory Animal Breeding Co., Ltd. or other qualified laboratory animal suppliers.

[0151] (3) Tumor inoculation: 0.2 mL (5 × 10⁻⁶) of the inoculated solution was injected into the tumor. 6 HCC1954 cells (PBS:Matrixgel = 1:1) were subcutaneously inoculated into the right posterior dorsal region of each mouse. In the efficacy experiment, the average tumor volume reached 100-150 mmHg. 3 Dosing will begin at the designated time for group assignment. Experimental group assignments and dosing regimens are shown in Table 9 below.

[0152] Table 9. Animal experimental grouping and dosing regimens:

[0153]

[0154]

[0155] Note:

[0156] 1. N: Number of mice in each group

[0157] 2. Dosage volume: 10 μL / g based on mouse body weight. If body weight decreases by more than 15%, the dosing regimen should be adjusted accordingly.

[0158] 3. QD: Once a day.

[0159] 4. Solvent (Vehicle): 10% dimethyl sulfoxide / 10% polyethylene glycol / 80% water

[0160] Animal grouping: Animals were weighed and tumor volume was measured before drug administration. Animals were then randomly assigned to groups based on tumor volume (randomized block design).

[0161] Experimental indicators: The experimental indicators are used to examine whether tumor growth is inhibited, delayed, or cured. The tumor diameter is measured twice a week (or every other day) 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.

[0162] The relative tumor-suppressive efficacy of the compounds was evaluated using TGI (%) or relative tumor proliferation rate T / C (%).

[0163] Data Analysis: The t-test is used for comparisons between two groups. One-way ANOVA is used for comparisons between three or more groups. If the F-values ​​show a significant difference, multiple comparisons should be performed after ANOVA analysis. All data analyses should be performed using SPSS or Graphpad Prism. A p-value < 0.05 is considered statistically significant.

[0164] 3. The experimental results are shown in Table 10.

[0165] Table 10 shows the efficacy results of compound (I) in the mouse HCC1954 in vivo pharmacodynamic model.

[0166]

[0167] Conclusion: Compound (I) exhibited dose-dependent tumor growth inhibition in the mouse HCC1954 in vivo pharmacodynamic model.

Claims

1. The A crystal form of the compound of formula (I), characterized in that... Its X-ray powder diffraction pattern exhibits characteristic diffraction peaks at the following 2θ angles: 5.25±0.20°, 7.78±0.20°, 8.74±0.20°, 10.46±0.20°, 14.99±0.20°, 16.25±0.20°, 17.50±0.20°, and 18.48±0.20°. 。 2. The A-type crystal according to claim 1, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.25±0.20°, 8.74±0.20°, 10.46±0.20°, 6.65±0.20°, 7.78±0.20°, 12.60±0.20°, 13.26±0.20°, 13.87±0.20°, 14.44±0.20°, 14.99±0.20°, 15.53±0.20°, 16.25±0.20°, 17.50±0.20°, 18.48±0.20°, 19.01 ±0.20°, 19.78±0.20°, 20.55±0.20°, 21.01±0.20°, 21.33±0.20°, 23.72±0.20°, 23.96±0.20°, 24.46±0.20°, 24.79±0.20°, 25.02±0.20°, 25.69±0.20°, 26.00±0.20°, 26.29±0.20°, 26.72±0.20°, 27.34±0.20°, 27.58±0.20°, 28.17±0.20° and 28.57±0.20°.

3. The A-type crystal according to claim 1, its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.2511°, 6.6532°, 7.7817°, 8.7418°, 10.4626°, 12.5958°, 13.2601°, 13.8718°, 14.4446°, 14.9927°, 15.5346°, 16.2492°, 17.5047°, 18.4752°. , 19.0069°, 19.7791°, 20.5468°, 21.0093°, 21.3325°, 23.7228°, 23.9609°, 24.4579°, 24.7921°, 25.0236°, 25.6910°, 26.0008°, 26.2926°, 26.7249°, 27.3381°, 27.5777°, 28.1708° and 28.5699°.

4. The A-type crystal according to any one of claims 1 to 3, its XRPD pattern is basically as shown in Figure 1.

5. The A-type crystal according to any one of claims 1 to 3, wherein the differential scanning calorimetry curve has an endothermic peak starting point at 90.5±3.0℃ and 181.7±3.0℃ respectively.

6. The A-type crystal according to any one of claims 1 to 3, its DSC spectrum is basically as shown in Figure 2.

7. The A crystal form according to any one of claims 1 to 3, wherein the thermogravimetric analysis curve shows a weight loss of 2.55% at 110.0 ± 3.0 °C.

8. The A crystal form according to any one of claims 1 to 3, its TGA spectrum is basically as shown in Figure 3.

9. The B crystal form of the compound of formula (I), characterized in that... Its X-ray powder diffraction pattern exhibits characteristic diffraction peaks at the following 2θ angles: 5.48±0.20°, 9.38±0.20°, 13.25±0.20°, 19.42±0.20°, 21.84±0.20°, and 24.90±0.20°. 。 10. The B-type crystal according to claim 9, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.4752°, 9.3771°, 10.9011°, 13.2511°, 16.3529°, 18.9134°, 19.4248°, 21.8375°, 24.8988°, 26.6143° and 27.6851°.

11. The B crystal form according to claim 9 or 10, the XRPD pattern of which is substantially as shown in Figure 4.

12. The B crystal form according to claim 9 or 10, wherein the differential scanning calorimetry curve has an endothermic peak starting point at 83.5 ± 3.0 °C.

13. The B crystal form according to claim 9 or 10, its DSC spectrum is essentially as shown in Figure 5.

14. The B crystal form according to claim 9 or 10, wherein the thermogravimetric analysis curve shows a weight loss of 6.73% at 135.0 ± 3.0 °C.

15. The B crystal form according to claim 9 or 10, its TGA spectrum is essentially as shown in Figure 6.

16. The use of crystal form A according to any one of claims 1 to 8 or crystal form B according to any one of claims 9 to 15 in the preparation of a medicament for treating breast cancer.

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