A crystalline form of a sulfonamide derivative and a process for its preparation

Abstract

The present disclosure relates to a crystalline form of a sulfonamide derivative and a method for preparing the same. In particular, the present disclosure provides a new crystalline form of a compound represented by formula (1) and a method for preparing the same.

Description

[0001] This application claims priority to Chinese patent application 2023101075697, filed on February 10, 2023. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This disclosure belongs to the field of pharmaceutical technology and relates to a crystalline form of a sulfonamide derivative and its preparation method. Background Technology

[0003] Lysine acetyltransferases (KATs) are a class of enzymes that catalyze the transfer of acetyl groups from acetyl-CoA to the ε-amino group of lysine residues in protein substrates. Lysine acetylation affects protein function, playing a crucial regulatory role in chromosome structure, gene transcription regulation, DNA binding capacity, enzyme activity and stability, protein-protein interactions, and intracellular localization. KATs are divided into several subfamilies, with MYST (MOZ, YBF2 / SAS3, SAS2, TIP60) being the largest, including KAT5 (TIP60), KAT6A (MOZ; MYST3), KAT6B (MORF; MYST4), KAT7 (HBO; MYST2), and KAT8 (MOF; MYST1). KAT6A / B, as key members of the MYST family, play a vital role in development, the maintenance of stem cells in hematopoiesis and the immune system, tumorigenesis and drug resistance.

[0004] Published patent applications for KAT6 inhibitors include WO2016198507A1, WO2019243491A1, WO2019043139A1, WO2019108824A1, WO2020216701A1, WO2020002587A1, WO2020254946A1, and WO2020254989A1, etc.

[0005] PCT / CN2022 / 111395 provides a KAT6 inhibitor with the chemical name N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-benzopyran[8,7-d]isoxazol-9-yl)-2,6-dimethoxybenzenesulfonamide, having the structure shown in Formula 1.

[0006]

[0007] The crystal form of a pharmaceutical active ingredient often affects its chemical stability. Different crystallization and storage conditions can lead to changes in the crystal structure of the compound, sometimes even resulting in other crystal forms. Generally, amorphous drug products lack regular crystal structures and often have other defects, such as poor product stability, fine crystals, difficulty in filtration, easy agglomeration, and poor flowability. Polymorphism of drugs places different requirements on product storage, production, and scale-up. Therefore, in-depth research on the crystal forms of the aforementioned compounds and the improvement of their various properties is essential. Summary of the Invention

[0008] This disclosure provides a novel crystal form of the compound shown in Formula 1, which has good stability and can be better applied in clinical practice.

[0009]

[0010] The A crystal form of the compound of Formula 1 provided in this disclosure has a characteristic peak at 11.288, 16.624, 18.251, 19.639, 22.547 and 26.085 in its X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0011] In some embodiments, the X-ray powder diffraction pattern of the A crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 11.288, 16.624, 18.251, 19.639, 21.140, 22.547, 23.439, 26.085, 26.497, and 27.050.

[0012] In some embodiments, the X-ray powder diffraction pattern of the A crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 11.288, 16.624, 17.312, 18.251, 19.639, 20.086, 21.140, 22.547, 23.439, 24.234, 26.085, 26.497, and 27.050.

[0013] In some embodiments, the X-ray powder diffraction pattern of the A crystal form of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 1 As shown.

[0014] This disclosure also provides a method for preparing the crystal form of compound A shown in Formula 1, wherein the method is selected from:

[0015] Method 1: Add the compound shown in Formula 1 to solvent (I) and slurry to precipitate crystals; wherein solvent (I) is selected from one or more of alcohol solvents, ketone solvents, ester solvents, ether solvents, hydrocarbon solvents, nitrile solvents, water, and dimethyl sulfoxide; wherein the alcohol solvent is selected from methanol, ethanol, isopropanol, n-propanol, benzyl alcohol, 1,2-propanediol, and isoamyl alcohol; wherein the ketone solvent is selected from acetone and methyl isobutyl ketone; wherein the ester solvent is selected from ethyl acetate and isopropyl acetate; wherein the ether solvent is selected from methyl tert-butyl ether, propylene glycol methyl ether, isopropyl ether, and tetrahydrofuran; wherein the hydrocarbon solvent is selected from nitromethane, n-heptane, cyclohexane, toluene, and p-xylene; and wherein the nitrile solvent is selected from acetonitrile.

[0016] Method 2: The compound of Formula 1 is heated and dissolved in solvent (II), and crystallized by stirring at room temperature or cooling; the solvent (II) is selected from one or more of acetonitrile, nitromethane, benzyl alcohol, N,N-dimethylacetamide, dimethyl sulfoxide, N-methylpyrrolidone, acetonitrile / methanol, and 1,2-dichloroethane.

[0017] Method 3: Dissolve the compound of Formula 1 in solvent (III), add solvent (IV) and stir to induce crystallization; wherein solvent (III) is selected from dimethyl sulfoxide or dichloromethane; wherein solvent (IV) is selected from one or more of water, methanol, ethanol, isopropanol, isopropyl ether, methyl tert-butyl ether, acetone, methyl isobutyl ketone, acetonitrile, ethyl acetate, and toluene.

[0018] This disclosure also provides a B-type crystal form of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 7.100, 11.249, 18.241, 22.580, and 26.107.

[0019] In some embodiments, the B crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.454, 7.100, 11.249, 18.241, 19.637, 20.092, 20.407, 22.580, 26.107, and 26.550.

[0020] In some embodiments, the X-ray powder diffraction pattern of the B crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 6.454, 7.100, 10.448, 11.249, 16.595, 18.241, 19.637, 20.092, 20.407, 22.580, 23.470, 26.107, and 26.550.

[0021] In some embodiments, the X-ray powder diffraction pattern of the B crystal form of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 2 As shown.

[0022] This disclosure also provides a method for preparing crystal form B, the method comprising:

[0023] The compound of Formula 1 is dissolved in solvent (V), and solvent (VI) is added and stirred to induce crystallization; wherein solvent (V) is selected from dimethyl sulfoxide; and solvent (VI) is selected from methyl tert-butyl ether.

[0024] This disclosure also provides a C-crystal form of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 6.484, 7.056, 7.907, 11.060, 14.223, and 20.342.

[0025] In some embodiments, the C-crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.484, 7.056, 7.907, 11.060, 14.223, 15.973, 20.342, 21.460, 23.090, and 26.398.

[0026] In some embodiments, the C-crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles of 2θ, exhibits characteristic peaks at 6.484, 7.056, 7.907, 11.060, 12.702, 14.223, 15.973, 17.526, 20.342, 21.460, 23.090, 26.398, and 27.500.

[0027] In some embodiments, the X-ray powder diffraction pattern of the C-crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, is as follows: Figure 3 As shown.

[0028] This disclosure also provides a method for preparing the C-type crystal, the method comprising:

[0029] Method 1: Dissolve the compound of Formula 1 in solvent (VII), and add solvent (VIII) to precipitate crystals; wherein solvent (VII) is selected from dichloromethane or tetrahydrofuran; wherein solvent (VIII) is selected from one or more of tetrahydrofuran and n-hexane.

[0030] Method 2: Dissolve the compound of Formula 1 in solvent (IX) and stir to induce crystallization; the solvent (IX) is selected from chloroform.

[0031] This disclosure also provides a D-crystal form of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 6.504, 7.051, 7.896, 11.086, 15.876 and 20.238.

[0032] In some embodiments, the D crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.504, 7.051, 7.896, 11.086, 13.560, 14.194, 15.876, 17.504, 18.599, 20.238, 22.244, and 23.963.

[0033] In some embodiments, the D crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.504, 7.051, 7.896, 11.086, 13.560, 14.194, 15.876, 17.504, 18.599, 20.238, 20.633, 22.244, 23.963, and 26.487.

[0034] In some embodiments, the X-ray powder diffraction pattern of the D crystal form of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 4 As shown.

[0035] This disclosure also provides a method for preparing a D-type crystal, the method comprising:

[0036] The compound of Formula 1 was dissolved in solvent (X) and stirred to crystallize; the solvent (X) was selected from N,N-dimethylformamide.

[0037] This disclosure also provides an E-crystal form of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 6.490, 7.069, 10.968, 14.194, 16.199 and 20.486.

[0038] In some embodiments, the E crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.490, 7.069, 8.034, 10.968, 14.194, 16.199, 18.656, 20.486, 21.539, and 22.949.

[0039] In some embodiments, the E crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.490, 7.069, 8.034, 10.968, 14.194, 16.199, 18.656, 20.486, 21.539, 22.360, 22.949, 23.979, 24.676, and 26.411.

[0040] In some embodiments, the X-ray powder diffraction pattern of the E crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, is as follows: Figure 5 As shown.

[0041] This disclosure also provides a method for preparing the E-crystal form, the method comprising:

[0042] The compound shown in Formula 1 is dissolved in solvent (XI) and stirred to crystallize; the solvent (XI) is selected from 1,2-dichloroethane.

[0043] This disclosure also provides an F-crystal form of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 7.327, 9.387, 13.968, 15.793, 20.198, and 21.813.

[0044] In some embodiments, the F crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles of 2θ, exhibits characteristic peaks at 7.327, 8.690, 9.387, 9.653, 12.583, 13.968, 15.793, 17.270, 20.198, 20.810, 21.813, 22.186, and 25.444.

[0045] In some embodiments, the F crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 7.327, 8.690, 8.970, 9.387, 9.653, 12.583, 13.968, 15.793, 16.429, 17.270, 17.975, 20.198, 20.810, 21.813, 22.186, 23.410, 24.438, 25.444, and 27.356.

[0046] In some embodiments, the X-ray powder diffraction pattern of the F crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, is as follows: Figure 6 As shown.

[0047] This disclosure also provides a method for preparing the F crystal form, the method comprising: dissolving the compound shown in Formula 1 in a solvent (XII) and stirring to induce crystallization; wherein the solvent (XII) is selected from ethyl acetate.

[0048] In some embodiments, the G crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles of 2θ, has characteristic peaks at 6.614, 7.192, 8.164, 11.113, 16.302, and 20.560.

[0049] In some embodiments, the G crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.614, 7.192, 8.164, 11.113, 11.377, 12.978, 14.360, 16.302, 20.560, 21.639, 22.588, 23.042, 24.186, and 26.521.

[0050] In some embodiments, the G crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 6.614, 7.192, 8.164, 11.113, 11.377, 12.266, 12.978, 13.738, 14.360, 16.302, 18.807, 20.560, 21.639, 22.588, 23.042, 24.186, 26.080, and 26.521.

[0051] In some embodiments, the X-ray powder diffraction pattern of the G crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, is as follows: Figure 7 As shown.

[0052] This disclosure also provides a method for preparing G crystal form, the method comprising: dissolving the compound shown in Formula 1 in a solvent (XIII), and evaporating the solvent to crystallize; wherein the solvent (XIII) is selected from dichloromethane.

[0053] This disclosure also provides an H-crystal form of the compound shown in Formula 1, with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 5.271, 10.587, 13.230, 16.017, 21.498, 22.753 and 26.938.

[0054] In some embodiments, the H crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 5.271, 10.587, 11.892, 13.230, 16.017, 16.625, 19.110, 21.498, 22.753, 24.080, 26.938, and 27.536.

[0055] In some embodiments, the H crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 5.271, 10.587, 11.892, 13.230, 16.017, 16.625, 19.110, 21.498, 22.753, 24.080, 25.459, 26.175, 26.938, 27.536, 31.694, and 38.116.

[0056] In some embodiments, the X-ray powder diffraction pattern of the H crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, is as follows: Figure 8 As shown.

[0057] This disclosure also provides a method for preparing the H-type crystal, the method comprising the following steps:

[0058] a) Mix the compound shown in Formula 1 with an aqueous sodium hydroxide solution;

[0059] b) Stirring at 37°C precipitates a solid;

[0060] c) Discard the supernatant, add hydrochloric acid-water solution, suspend and slurry at 37℃, centrifuge and vacuum dry to obtain solid.

[0061] In some embodiments, the I crystal form of the compound shown in Formula 1, as expressed in X-ray powder diffraction patterns at diffraction angles 2θ, has characteristic peaks at 5.963, 11.883, 13.953, 19.518, 20.305, and 22.086.

[0062] In some embodiments, the X-ray powder diffraction pattern of the I crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.963, 9.438, 10.303, 11.883, 13.953, 17.945, 18.356, 19.518, 19.807, 20.305, 22.086, 25.009, and 29.576.

[0063] In some embodiments, the X-ray powder diffraction pattern of the I crystal form of the compound shown in Formula 1, expressed as a diffraction angle 2θ, has characteristic peaks at 5.963, 9.438, 10.303, 11.156, 11.883, 13.446, 13.953, 17.945, 18.356, 19.518, 19.807, 20.305, 21.559, 22.086, 25.009, 29.576, and 30.192.

[0064] In some embodiments, the X-ray powder diffraction pattern of the I crystal form of the compound shown in Formula 1, expressed in terms of the diffraction angle 2θ, is as follows: Figure 9 As shown.

[0065] This disclosure also provides a method for preparing crystal form I, the method comprising: dissolving the compound shown in Formula 1 in a solvent (XIV), and evaporating the solvent to crystallize; wherein the solvent (XIV) is selected from dichloromethane.

[0066] This disclosure also provides a pharmaceutical composition comprising the aforementioned crystal form A, B, C, D, E, F, G, H, or I, and a pharmaceutical excipient optionally selected from pharmaceutically acceptable excipients.

[0067] This disclosure also provides a pharmaceutical composition prepared from the aforementioned crystal form A, B, C, D, E, F, G, H, or I, and optionally a pharmaceutically acceptable excipient.

[0068] This disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned crystal form A, B, C, D, E, F, G, H, or I with a pharmaceutically acceptable excipient.

[0069] This disclosure also provides the use of the aforementioned crystal forms A, B, C, D, E, F, G, H, or I, or the aforementioned compositions, in the preparation for the prevention and / or treatment of cancer.

[0070] The uses described in this disclosure, wherein the cancers are selected from breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, gastric cancer, colorectal cancer, lung cancer, kidney cancer, liver cancer, cervical cancer, endometrial cancer, myeloma, leukemia, lymphoma, acoustic neuroma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, bronchial cancer, sarcoma, chordoma, choriocarcinoma, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, hemangioendothelioma, ependymoma, epithelial carcinoma, esophageal cancer, essential thrombocytosis, Ewing's tumor, testicular cancer, glioma, heavy chain disease. Hemangioblastoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, neuroblastoma, midline carcinoma of NUT, glioma, bone cancer, nasopharyngeal carcinoma, oral cancer, thyroid cancer, pineal tumor, polycythemia vera, retinoblastoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, synovoma, sweat gland carcinoma, Waldenström macroglobulinemia, and Wilms' tumor; preferably, the cancers are selected from breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, gastric cancer, colorectal cancer, and lung cancer.

[0071] The "2θ or 2θ angle" mentioned in this disclosure refers to the diffraction angle, where θ is the Bragg angle, and the unit is ° or degree; the error range of 2θ for each characteristic peak is +0.20 (including the case where the number has more than one decimal place after rounding), specifically -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

[0072] The numerical values ​​in this disclosure, such as those relating to the content of certain substances, are calculated data and inevitably contain a certain degree of error. Generally, ±10% is within the reasonable error range. The error may vary to some extent depending on the context in which it is used, but this variation shall not exceed ±10%, and may be ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%, preferably ±5%.

[0073] The starting material used in the crystal form preparation method disclosed herein can be any form of compound, including but not limited to: amorphous, arbitrary crystal form, hydrate, solvate, etc.

[0074] The drying temperature described in this disclosure is generally 25℃-100℃, preferably 40℃-70℃, and can be dried under normal pressure or reduced pressure.

[0075] The crystallization methods described in this disclosure include room temperature crystallization, cooling crystallization, solvent evaporation crystallization, and seed crystallization induction. The cooling temperature is selected from below 65°C, preferably from -10°C to 60°C. Stirring can also be performed during the crystallization process.

[0076] The “differential scanning calorimetry or DSC” described in this disclosure refers to measuring the temperature difference and heat flow difference between the sample and the reference material during the sample heating or isothermal process, in order to characterize all physical and chemical changes related to thermal effects and obtain phase transition information of the sample.

[0077] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "Guiding Principles on Hygroscopicity of Drugs" in Part IV of the 2015 edition of the Chinese Pharmacopoeia,

[0078] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0079] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15%;

[0080] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15% but not less than 2%;

[0081] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0082] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0083] The “excipients” described in this disclosure include, but are not limited to, any adjuvants, carriers, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, or emulsifiers that have been approved by the U.S. Food and Drug Administration for use in humans or livestock. Attached Figure Description

[0084] Figure 1 The image shows the XRPD spectrum of crystal form A of compound 1.

[0085] Figure 2 The image shows the XRPD spectrum of crystal form B of compound 1.

[0086] Figure 3 The image shows the XRPD spectrum of crystal form C of compound 1.

[0087] Figure 4 The image shows the XRPD spectrum of crystal form D of compound 1.

[0088] Figure 5 The image shows the XRPD spectrum of crystal form E of compound 1.

[0089] Figure 6 The image shows the XRPD spectrum of crystal form F of compound 1.

[0090] Figure 7 The image shows the XRPD spectrum of crystal form G of compound 1.

[0091] Figure 8 The image shows the XRPD spectrum of crystal form H of compound 1.

[0092] Figure 9 The image shows the XRPD spectrum of crystal form I of compound 1. Detailed Implementation

[0093] The present disclosure will be explained in more detail below with reference to embodiments or experimental examples. The embodiments or experimental examples in the present disclosure are only used to illustrate the technical solutions in the present disclosure and are not intended to limit the substance and scope of the present disclosure.

[0094] Test conditions of the instruments used in the experiment:

[0095] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ ohms. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE NEO 500M NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0096] MS measurements were performed using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS LC-MS system (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD system (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive system (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0097] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent HPLC 1200DAD, an Agilent HPLC 1200VWD, and a Waters HPLC e2695-2489 HPLC system.

[0098] High performance liquid chromatography (HPLC) was performed using Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson-281 preparative chromatographs.

[0099] Silica gel column chromatography generally uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0100] XRPD (X-ray Powder Diffraction) was used for analysis: measurements were performed using a BRUKER D8 X-ray diffractometer. Specific data collected included: Cu anode (40 kV, 40 mA), Cu-Kα1 rays. Kα2 rays Kβ rays Scanning mode: θ / 2θ, scanning range (2θ range): 5°~45°.

[0101] DSC stands for Differential Scanning Calorimetry: Measurements were performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10℃ / min. The specific temperature range was referenced from the corresponding spectra (mostly 25-350℃), and the nitrogen purging rate was 50mL / min.

[0102] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer, with a heating rate of 10℃ / min, and the specific temperature range was referenced from the corresponding spectrum (mostly 25-350℃). The nitrogen purging rate was 50mL / min.

[0103] DVS stands for Dynamic Moisture Adsorption: Detection is performed using SMS DVS Advantage at 25℃, with humidity changes of 50%-95%-0%-95%-50%, in 10% increments (the final step is 5%) (the specific humidity range is subject to the corresponding spectrum; the methods listed here are the most commonly used). The judgment criterion is that dm / dt is not greater than 0.002%.

[0104] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, and Darui Chemicals.

[0105] Unless otherwise specified in the examples, all reactions can be carried out under an argon or nitrogen atmosphere.

[0106] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0107] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system used for column chromatography to purify the compounds, and the developing solvent system for TLC included: A: n-hexane / ethyl acetate system, B: dichloromethane / methanol system. The volume ratio of the solvent was adjusted according to the polarity of the compounds, and small amounts of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0108] Example 1: Preparation of Compound 1

[0109] N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazo-9-yl)-2,6-dimethoxybenzenesulfonamide

[0110]

[0111] first step

[0112] 4-Bromo-2-((2,4-dimethoxybenzyl)oxy)-6-fluorobenzonitrile 1b

[0113] 4-Bromo-2,6-difluorobenzonitrile 1a (22 g, 101 mmol, Shaoyuan) and 2,4-dimethoxybenzyl alcohol (18.5 g, 110 mmol, Biotin) were dissolved in N,N-dimethylformamide (200 mL), and cesium carbonate (49 g, 150 mmol, Shaoyuan) was added. The reaction mixture was stirred at 60 °C for 16 hours. The reaction mixture was cooled to room temperature, filtered under reduced pressure, and the filtrate was diluted with ethyl acetate (500 mL). The filtrate was washed with saturated sodium chloride solution (30 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give title product 1b (36.9 g, yield: 100%). This product was used directly in the next reaction without purification.

[0114] Step 2

[0115] 4-Bromo-2-fluoro-6-hydroxybenzonitrile 1c

[0116] Compound 1b (36.9 g, 100.7 mmol) was dissolved in dichloromethane (250 mL), cooled to 0 °C, and trifluoroacetic acid (39 g, 342 mmol, Adamas) was added dropwise. The reaction mixture was then stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using eluent system A to give the title product 1c (9.7 g, yield: 44.5%).

[0117] Step 3

[0118] 2-(allyloxy)-4-bromo-6-fluorobenzonitrile 1d

[0119] Compound 1c (10.7 g, 49.5 mmol) was dissolved in N,N-dimethylformamide (120 mL). The reaction mixture was cooled to 0 °C, and cesium carbonate (24 g, 73.7 mmol, BIDE) and allyl bromide (11.2 g, 92.6 mmol, Adamas) were added. The reaction mixture was then stirred at room temperature for 4 hours. The reaction mixture was filtered under reduced pressure, and the filtrate was diluted with ethyl acetate (400 mL). The filtrate was washed with saturated sodium chloride solution (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system A to give the title product 1d (11.7 g, yield: 92%). 1 H NMR (500MHz, CDCl3) δ7.02 (dt, 1H), 6.95 (t, 1H), 6.04 (m, 1H), 5.57-5.47 (m, 1H), 5.41 (dt, 1H), 4.75-4.64 (m, 2H).

[0120] Step 4

[0121] 3-Allyl-4-bromo-6-fluoro-2-hydroxybenzonitrile 1e

[0122] Compound 1d (3.35 g, 13.1 mmol) was dissolved in 1,2-dichlorobenzene (80 mL), and nitrogen gas was purged three times. The reaction mixture was stirred at 180 °C for 13 hours. After cooling to room temperature, the residue was purified by silica gel column chromatography (wet loading) with eluent system A to give the title product 1e (2.77 g, yield: 82.7%). 1 H NMR (500MHz, CDCl3) δ7.07 (dd, 1H), 6.45 (s, 1H), 5.90 (dddd, 1H), 5.24-5.10 (m, 2H), 3.68-3.55 (m, 2H).

[0123] Step 5

[0124] 4-Bromo-6-fluoro-2-hydroxy-3-(3-hydroxypropyl)benzonitrile 1f

[0125] Compound 1e (4.8 g, 18.7 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL), and borane tetrahydrofuran solution (1.0 M, 22 mL, 22 mmol, Adamas) was added dropwise at 0 °C. The reaction mixture was stirred in an ice bath for 2 hours. Then, 3 M sodium hydroxide aqueous solution (13 mL, 39 mmol) and 30% hydrogen peroxide (3.0 mL) were added sequentially in an ice bath, followed by stirring for 10 min. The pH of the reaction mixture was adjusted to 2 with 2 M hydrochloric acid, and the mixture was extracted with ethyl acetate (100 mL × 2). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system A to give the title product 1f (3.5 g, yield: 68.1%). MS m / z (ESI): 275.8 [M+1]. 1 H NMR (500MHz, CDCl3) δ7.04 (d, 1H), 3.71 (t, 2H), 3.00-2.98 (m, 2H), 2.01-1.96 (m, 2H).

[0126] Step 6

[0127] 1g of 5-bromo-7-fluorotryptane-8-carboxynitrile

[0128] Compound 1f (3.8 g, 13.9 mmol) was dissolved in anhydrous tetrahydrofuran (80 mL). The reaction solution was cooled to 0 °C, and triphenylphosphine (4.4 g, 16.8 mmol, Sinopharm) and diisopropyl azodicarbonate (3.4 g, 16.8 mmol, Shaoyuan) were added. The reaction solution was then heated to room temperature and stirred for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using eluent system A to give the title product 1 g (3.0 g, yield: 84.5%). 1 H NMR (500MHz, CDCl3) δ7.03 (d, 1H), 4.33 (t, 2H), 2.77-2.74 (m, 2H), 2.12-2.08 (m, 2H). MS m / z(ESI): 257.8[M+1].

[0129] Step 7

[0130] 8-Cyano-7-Fluorostrene-5-carboxylic acid methyl ester 1h

[0131] 1 g (2.6 g, 10.2 mmol) of the compound was dissolved in 40 mL of a mixed solvent of methanol and N,N-dimethylformamide (V:V = 1:3). Then, 1,1′-bis(diphenylphosphine)ferrocene palladium dichloride (800 mg, 1.09 mmol, Adamas) and triethylamine (3.0 g, 2.93 mmol, Sinopharm) were added sequentially. The mixture was purged with carbon monoxide three times, and stirred at 10 bar and 90 °C for 16 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, diluted with ethyl acetate (150 mL), and washed with saturated sodium chloride solution (50 mL × 3). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system A to give the title product 1h (2.1 g, yield: 87.9%). 1 H NMR (500MHz, CDCl3) δ7.26 (d, 1H), 4.38-4.36 (m, 2H), 3.93 (s, 3H), 3.10-3.07 (m, 2H), 2.08-2.03 (m, 2H). MS m / z(ESI): 235.9[M+1].

[0132] Step 8

[0133] 7-Fluoro-5-(hydroxymethyl)trypanone-8-carboxynitrile 1i

[0134] Compound 1h (2.1 g, 8.93 mmol) was dissolved in dry tetrahydrofuran (40 mL), purged with nitrogen three times, and the reaction solution was cooled to 0 °C. Lithium borohydride (2 M, 18 mmol, 9.0 mL, Adamas) was added. The reaction solution was heated to 70 °C and stirred for 2 hours. The reaction solution was cooled to room temperature, quenched with water (1 mL), diluted with ethyl acetate (100 mL), and washed with saturated sodium chloride solution (50 mL × 2). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system A to give the title product 1i (1.84 g, yield: 99.5%). MS m / z (ESI): 207.9 [M+1].

[0135] Step 9

[0136] 5-((1H-pyrazol-1-yl)methyl)-7-fluorotryptane-8-carboxynitrile 1k

[0137] Compound 1i (1.8 g, 8.69 mmol) and 1-(methanesulfonyl)-1H-pyrazole 1j (1.5 g, 10.3 mmol, prepared by the method disclosed in Scheme 8 Intermediate 13 on page 63 of patent application "WO2020254946A1") were dissolved in acetonitrile (30 mL), and cesium carbonate (4.2 g, 12.9 mmol) was added. The mixture was reacted at 70 °C for 1 hour. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using eluent system A to give the title product 1k (1.9 g, yield: 85.0%). MS m / z (ESI): 258.0 [M+1].

[0138] Step 10

[0139] 5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-amine

[0140] Compound 1k (1.9 g, 7.39 mmol) and acetyloxyoxime acid (1.7 g, 22.2 mmol, Adamas) were dissolved in N,N-dimethylformamide (30 mL) and water (4.0 mL), and potassium carbonate (6.2 g, 44.9 mmol, Sinopharm) was added. The reaction mixture was stirred at 70 °C for 24 hours. The reaction mixture was cooled to room temperature, water (100 mL) was added, the mixture was filtered, the filter cake was collected and dried to give the title product 1l (1.65 g, yield: 82.7%). 1H NMR (500MHz, DMSO-d6) δ7.78 (d, 1H), 7.52 (d, 1H), 6.34 (s, 1H), 6.32 (t, 1H), 5.85 (s, 2H), 5.39 (s, 2H), 4.25-4.23 (m, 2H), 2.68 (t, 2H), 2.03-1.98 (m, 2H). MS m / z(ESI): 271.0[M+1].

[0141] Step 11

[0142] N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazo-9-yl)-2,6-dimethoxybenzenesulfonamide

[0143] Compound 1l (200 mg, 0.740 mmol) and compound 1m (300 mg, 1.27 mmol) were dissolved in pyridine (5.0 mL), and nitrogen was purged three times. The reaction solution was microwaved at 120 °C for 3 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was purified by high performance liquid chromatography (Xtimate phenyl-hexyl Prep C18 5 μm, 30 × 150 mm; mobile phase: A-aqueous phase (0.1% ammonia): B-acetonitrile = 5%-45% (20 min), flow rate: 30 mL / min) to give title product 1 (40 mg, yield: 11.5%). 1 H NMR (500MHz, DMSO-d6) δ9.40 (s, 1H), 7.79 (d, 1H), 7.52 (d, 1H), 7.48 (t, 1H), 6.77 (d, 2H), 6.4 3(s, 1H), 6.32(t, 1H), 5.42(s, 2H), 4.25(t, 2H), 3.78(s, 6H), 2.70(t, 2H), 2.04-1.99(m, 2H). MS m / z(ESI): 470.8[M+1].

[0144] Test Example 1: Detection of KAT6 enzyme activity in compound 1 (AlphaScreen method)

[0145] I. Experimental Materials

[0146] 1. KAT6A (Custom-made by Chempartner)

[0147] 2. Ovalbumin (Sigma-Aldrich, A5378-5G)

[0148] 3. 2M Tris-HCl solution, pH 7.8, sterile (Sangon Biotech B548140-0500)

[0149] 4. 5M NaCl solution (Sangon Biotech, B548121-0100)

[0150] 5.EDTA (0.5M), pH 8.0, RNase-free (Thermofisher, AM9260G)

[0151] 6. Tween-20 (Biotech, A100777-0500)

[0152] 7.DTT, 1M (Invitrogen, P2325)

[0153] 8. Acetyl-CoA (CAYMAN, Cat. No. 16160)

[0154] 9. Biotin-labeled recombinant histone H3.1 (human) (Active Motif 31696)

[0155] 10. 384-well plate, light gray (Perkin Elmer, Cat. No. 6005350)

[0156] 11. Astragalus acid (MCE, Cat. No. HY-N2020)

[0157] 12. AlphaScreen Streptavidin Donor beads 5mg (PerkinElmer, 6760002)

[0158] 13. AlphaScreen Protein A Acceptor beads, 5 mg (PerkinElmer, 6760137M)

[0159] 14. Acetylated-Lysine Antibody (CST 9441S)

[0160] 15. Phera Star microplate reader (BMG Labtech)

[0161] II. Experimental Methods

[0162] 1. Reagent preparation

[0163] a. 1× Detection buffer: 100mM Tris-HCl, pH 7.8; 15mM NaCl; 1mM EDTA; 0.01% Tween-20; 1mM DTT; 0.01% m / v ovalbumin.

[0164] b. KAT enzyme solution: Prepare 1× detection buffer to a final concentration of 1.25 nM.

[0165] c. Ac-CoA and H3 mixed substrate: Prepare a mixed substrate of Ac-CoA with a final concentration of 1000 nM and H3 with a final concentration of 55 nM using 1× detection buffer.

[0166] d. Compounds: Starting concentration 100 μM, 3-fold dilution, 10 concentration gradients. All compounds were diluted 83-fold with 1× detection buffer and set aside.

[0167] e. Detection reagents: 1× detection buffer to prepare AlphaScreen protein A receptor beads with a final concentration of 8 ng / μL, AlphaScreen streptavidin donor beads with a final concentration of 8 ng / μL, acetylated lysine antibody diluted 1:1500, and 100 μM astaxanthin.

[0168] 2. Experimental Procedure

[0169] Add 3 μL of the prepared enzyme solution to each well of a 384-well plate. Add 3 μL of 1× detection buffer to wells 23 and 24 (Min). Add 3 μL of compound solution to each well, and 3 μL of buffer to each well in the Min column. Add 3 μL of DMSO solution to each well in the 1st and 2nd columns (Max) as a control. Centrifuge, mix and vortex for 2 minutes; incubate at room temperature for 15 minutes. Add 6 μL of Ac-CoA and H3 mixed substrate to each well, centrifuge, mix and vortex for 2 minutes, and incubate at room temperature for 20 minutes. Add 6 μL of detection reagent to each well, centrifuge, mix and vortex for 2 minutes, and incubate at room temperature for 120 minutes in the dark. Read the plate using a microplate reader and record the AlphaScreen count values. Plot the graph using Graphpad software and calculate the IC50 of the compound. 50 The value is IC. 50 =0.3nM

[0170] Conclusion: Compound of Formula 1 has a good inhibitory effect on KAT6A.

[0171] Test Example 2: Compound 1's resistance to ZR-75-1 proliferation experiment

[0172] I. Reagents and Instruments

[0173] 1. ZR-75-1 (ATCC CRL1500)

[0174] 2. 1640 medium (Gibco, 22400-089)

[0175] 3. 0.25% trypsin-EDTA (1x) (Gibco, 25200-072)

[0176] 4. Penicillin-Streptomycin (Gibco, 15140-122)

[0177] 5.DPBS(1×)(Gibco, 14190-144)

[0178] 6. FBS (Gibco, 10091148)

[0179] 7. 96-well transparent black detection plate at the bottom (Coming, 3603)

[0180] 8. 96-well non-treated round bottom dispensing plate (JET BIOFIL, TCP-002-096)

[0181] 9. CellTiter-Glo buffer (Promega, G756B)

[0182] 10. CellTiter-Glo substrate (Promega, G755B)

[0183] 11. Automated Cell Counter (Countstar, IC 1000)

[0184] 12. Thermo Incubator (Thermo, I160)

[0185] 13.PHERAstar FS (BMG labtech, PHERAstar FS)

[0186] II. Experimental Methods

[0187] 1. Cell plating (Day 0)

[0188] a. Observe the cell state under a microscope to ensure that the cell confluence is around 90%.

[0189] b. Discard the cell supernatant, rinse once with DPBS, and discard the DPBS. Add an appropriate amount of trypsin to digest the cells, and incubate at 37°C for 5 minutes.

[0190] c. Stop digestion with an equal volume of 1640 medium containing 10% FBS, and collect the cell suspension. Centrifuge at 300g for 3 minutes. Resuspend the cells in an appropriate amount of fresh medium.

[0191] d. Take the resuspended cell suspension and count the cells.

[0192] e. Dilute the cell suspension to 5×10⁴ / mL with 1640 medium containing 10% FBS, 50 μL / well. ZR-75-1 is 2500 cells / well.

[0193] f. Incubate the cell culture plates overnight in an incubator at 37°C with 5% carbon dioxide.

[0194] 2. Administer medication (Day 1)

[0195] a. Dilute each compound to 9 concentration points using DMSO (starting concentration 100 μM, 3-fold dilution; for different compounds, follow IC50 concentrations). 50 (Different maximum concentrations can be adjusted accordingly). For example, in a 96-well round-bottom plate, 3 μL of the compound is serially diluted to 6 μL of DMSO.

[0196] b. Dilute each compound at each concentration point by 500 times into the corresponding volume of 1640 mL culture medium.

[0197] c. Add 50 μL of the diluted compound solution to each well of the cell plate containing 50 μL of cell supernatant.

[0198] d. Place the cell plate after drug administration in an incubator at 37°C with 5% carbon dioxide.

[0199] 3. Re-digest the plaster and add medication (Day 7)

[0200] a. Six days after drug administration, discard the drug-containing culture medium, then add 150 μL of DPBS per well to rinse once, and immediately remove the DPBS.

[0201] b. Add 50 μL of trypsin to digest the cells, incubate at 37°C for 3 minutes, and then add 1640 medium containing 10% FBS at 150 μL / well to terminate the digestion.

[0202] c. Mix the cells by pipetting with a pipette and re-coat the cells at a ratio of 1:8, i.e., aspirate 25 μL of cell suspension into a new 96-well plate (pre-added with 25 μL of 1640 medium containing 10% FBS).

[0203] d. Prepare and add the compound according to steps a to c in section 2, 50 μL per well.

[0204] e. Place the cell plate after drug administration in an incubator at 37°C with 5% carbon dioxide for incubation.

[0205] 4. CTG test (day 14)

[0206] a. Before use, allow the CellTiter-Glo buffer and lyophilized CellTiter-Glo substrate to equilibrate to room temperature, then mix them thoroughly to prepare 100 mL of CellTiter-Glo reagent (or remove the mixed CellTiter-Glo reagent from -20°C and equilibrate to room temperature).

[0207] b. Remove the plate to be tested from the incubator, equilibrate to room temperature, and add 50 μL of CellTiter-Glo reagent to each well.

[0208] c. Shake and mix for 2 minutes to allow the cells to fully lyse.

[0209] d. After the signal stabilizes at room temperature for 28 minutes, perform the detection on the Pherastar FS.

[0210] Table 1 shows the IC50 values ​​of compound 1 for inhibiting the proliferation of ZR-75-1. 50 Value and maximum inhibition rate

[0211]

[0212] Conclusion: Compound of Formula 1 has a good inhibitory effect on the proliferation of ZR-75-1.

[0213] Example 2: Preparation of crystal form A of the compound shown in Formula 1

[0214] 10 mg of the compound shown in Formula 1 was added to 1 mL of solvent, and the mixture was stirred and slurried at 60 °C - 5 °C with a heating / cooling rate of +0.75 °C / min. The mixture was filtered, the filter cake was collected, and vacuum dried to obtain a solid. X-ray powder diffraction analysis identified the product as crystal form A. The solvents used are shown in Table 2 below. The XRPD spectrum is shown below. Figure 1 As shown in Table 3, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peak has a peak value of 240.55℃. The TGA spectrum shows that there is essentially no weight loss between 30 and 267℃.

[0215] Table 2 Solvent selection and XRPD detection results for A-type preparation by pulping and crystallization.

[0216]

[0217] Table 3. XRPD characteristic diffraction peak data for crystal form A

[0218]

[0219]

[0220] Example 3: Preparation of crystal form A of the compound shown in Formula 1

[0221] 10 mg of the compound shown in Formula 1 was added to 1 mL of solvent, stirred at 60 °C until dissolved, cooled and stirred to crystallize, centrifuged, and then dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was of crystal form A.

[0222] Table 4. Solvent selection and XRPD detection results for A-type crystallization prepared by cooling crystallization.

[0223]

[0224] Example 4: Preparation of crystal form A of the compound shown in Formula 1

[0225] 10 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of dimethyl sulfoxide. After adding 1 mL of solvent, the precipitate was formed. The precipitate was then centrifuged and dried under vacuum to obtain a solid. The crystalline form of the obtained solid was determined by X-ray powder diffraction, as shown in Table 5 below.

[0226] Table 5. Solvent selection and XRPD detection results for A-type crystals prepared by dissolution.

[0227]

[0228] Example 5: Preparation of crystal form A of the compound shown in Formula 1

[0229] 10 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of dimethyl sulfoxide. After adding 1 mL of acetone, no precipitation occurred. The mixture was cooled to 5 °C and stirred to induce crystallization. After centrifugation, the solid was dried under vacuum. X-ray powder diffraction analysis showed that the product was in crystal form A.

[0230] Example 6: Preparation of crystal form A of the compound shown in Formula 1

[0231] 10 mg of the compound shown in Formula 1 was dissolved in 0.3 mL of dichloromethane. After adding 1 mL of solvent, the precipitate was formed. The precipitate was then centrifuged and dried under vacuum to obtain a solid. The crystalline form of the obtained solid was determined by X-ray powder diffraction, as shown in Table 6 below.

[0232] Table 6. Solvent selection and XRPD detection results for A-type crystals prepared by dissolution.

[0233]

[0234] Example 7: Preparation of crystal form B of the compound shown in Formula 1

[0235] 10 mg of the compound shown in Formula 1 was dissolved in 0.2 mL of dimethyl sulfoxide, and 1 mL of methyl tert-butyl ether was added. After stirring and slurry preparation at room temperature, the precipitate was obtained, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction analysis identified the product as crystal form B. The XRPD spectrum is shown below. Figure 2As shown in Table 7, the positions of its characteristic peaks are as follows. The DSC spectrum shows endothermic peaks at 79.87℃ and 240.13℃. The TGA spectrum shows a weight loss of 2.43% between 30-130℃.

[0236] Table 7. XRPD characteristic diffraction peak data for crystal form B.

[0237]

[0238]

[0239] Example 8: Preparation of crystal form C of the compound shown in Formula 1

[0240] 10 mg of the compound shown in Formula 1 was dissolved in 0.3 mL of dichloromethane, and 1 mL of tetrahydrofuran was added. The mixture was cooled to 5 °C, stirred, and stirred to induce crystallization. After centrifugation, the solid was dried under vacuum. X-ray powder diffraction analysis identified the product as crystal form C. The XRPD spectrum is shown below. Figure 3 As shown in Table 8, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peaks have peak values ​​of 99.14℃ and 240.84℃. The TGA spectrum shows a weight loss of 10.28% between 30-130℃.

[0241] Table 8. XRPD characteristic diffraction peak data for crystal form C.

[0242]

[0243] Example 9: Preparation of crystal form C of the compound shown in Formula 1

[0244] 10 mg of the compound shown in Formula 1 was added to 0.3 mL of chloroform, stirred at 60 °C until dissolved, cooled and stirred to crystallize, centrifuged, and then dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was in the C-form.

[0245] Example 10: Preparation of crystal form D of the compound shown in Formula 1

[0246] 10 mg of the compound shown in Formula 1 was added to 0.25 mL of N,N-dimethylformamide and stirred at 60 °C until dissolved. The solution was then cooled and stirred to induce crystallization. After centrifugation, the solid was dried under vacuum. X-ray powder diffraction analysis identified the product as crystal form D. The XRPD spectrum is shown below. Figure 4 As shown in Table 9, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peaks have peak values ​​of 104.54℃ and 240.90℃. The TGA spectrum shows a weight loss of 11.10% between 30 and 200℃.

[0247] Table 9. XRPD characteristic diffraction peak data for crystal form D.

[0248]

[0249] Example 11: Preparation of crystal form E of the compound shown in Formula 1

[0250] 10 mg of the compound shown in Formula 1 was added to 0.6 mL of 1,2-dichloroethane and stirred at 60 °C until dissolved. The solution was then cooled and stirred to induce crystallization. After centrifugation, the solid was dried under vacuum. X-ray powder diffraction analysis identified the product as crystal form E. The XRPD spectrum is shown below. Figure 5 As shown in Table 10, the positions of its characteristic peaks are as follows. The DSC spectrum shows endothermic peaks at 113.17℃ and 241.21℃. The TGA spectrum shows a weight loss of 11.13% between 30-130℃.

[0251] Table 10 XRPD characteristic diffraction peak data for crystal form E

[0252]

[0253] Example 12: Preparation of crystal form F of the compound shown in Formula 1

[0254] The compound of formula 1 (11 g, 23.4 mmol) was dispersed in 55 mL of ethyl acetate, stirred at room temperature for 1 hour, filtered, and the solid was collected and dried under vacuum at 40 °C for 6 hours to obtain the solid. X-ray powder diffraction analysis identified the product as crystal form F, and the XRPD spectrum is shown below. Figure 6 As shown in Table 11, the positions of its characteristic peaks are as follows. The DSC spectrum shows the endothermic peaks at 111.60℃, 115.04℃, and 241.06℃. The TGA spectrum shows a weight loss of 9.12% from 40℃ to 180℃.

[0255] Table 11 XRPD characteristic diffraction peak data for crystal form F

[0256]

[0257]

[0258] Example 13: Preparation of crystal form G of the compound shown in Formula 1

[0259] Compound 1 (216 mg, 0.46 mmol) was dispersed in 10 mL of dichloromethane, stirred at room temperature until dissolved, concentrated under reduced pressure and the solid was retained. The solid was then dried under vacuum at room temperature for 2 hours. X-ray powder diffraction analysis identified the product as crystal form G, and its characteristic peak positions are shown in Table 12. The X-ray powder diffraction pattern is shown below. Figure 7 As shown. The DSC spectrum shows: endothermic peaks at 87.44℃ and 239.07℃; exothermic peaks at 113.64℃ and 294.50℃. The TGA spectrum shows: weight loss of 2.51% from 40℃ to 150℃, and weight loss of 6.48% from 150℃ to 280℃.

[0260] Table 12 XRPD characteristic diffraction peak data for crystal form G

[0261]

[0262] Example 14: Preparation of crystal form H of the compound shown in Formula 1

[0263] 100 mg of the compound shown in Formula 1 was dissolved in 2 mL of 2M sodium hydroxide aqueous solution and 3 mL of pure water. The mixture was stirred at 37°C for 1 hour to precipitate a solid. The supernatant was discarded by centrifugation, and then 5 mL of 0.1M hydrochloric acid aqueous solution was added to suspend the solid at 37°C. After centrifugation and vacuum drying, the solid was obtained. X-ray powder diffraction analysis identified the product as crystal form H. The XRPD spectrum is shown below. Figure 8 As shown in Table 13, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peak has a peak value of 240.45℃. The TGA spectrum shows that the weight loss is 0.20% from 30 to 233℃.

[0264] Table 13 XRPD characteristic diffraction peak data for crystal form H

[0265]

[0266] Example 15: Preparation of Crystal Form I of the Compound Shown in Formula 1

[0267] Compound of Formula 1 (14 mg, 0.03 mmol) was dissolved in 0.5 mL of dichloromethane, filtered, and the filtrate was collected. After standing at room temperature for 120 hours, a solid precipitated. X-ray powder diffraction analysis identified the product as crystal form I, and its characteristic peak positions are shown in Table 14. The X-ray powder diffraction pattern is shown below. Figure 9 As shown. The DSC spectrum shows: endothermic peaks at 77.76℃ and 241.38℃; exothermic peaks at 113.29℃. The TGA spectrum shows: weight loss of 7.58% from 40℃ to 200℃.

[0268] Table 14 XRPD characteristic diffraction peak data for crystal form I

[0269]

[0270]

[0271]

[0272] Example 16: Hygroscopicity Study of Crystal Forms

[0273] Using Surface Measurement Systems intrinsic DVS, at 25℃, the humidity range was 0%-95%, with a step size of 10%. The judgment criterion was that the mass change dM / dT for each gradient was less than 0.002%, TMAX 360min, and two cycles were performed.

[0274] Table 15

[0275]

[0276] Example 17: Stability Study of Factors Affecting Crystal Form

[0277] The free-state samples were laid flat in the open and the stability of the samples was investigated under light (4500 Lux), high temperature (40℃, 60℃), and high humidity (RH 75%, RH 92.5%) conditions. The sampling period was 30 days.

[0278] Table 16

[0279]

[0280] Experimental results on influencing factors show that crystal form A has good physical and chemical stability under high temperature, high humidity, and light.

[0281] Example 18: Long-term / accelerated stability of crystal form

[0282] The free-state sample was sealed in an aluminum foil bag and its stability was investigated under conditions of 25℃ / 60%RH and 40℃ / 75%RH, respectively. The results are shown below.

[0283] Table 17

[0284]

[0285] Experimental results show that crystal form A exhibits good physical and chemical stability after being placed under long-term accelerated conditions for 9 months.

Claims

1. A crystal form A of the compound shown in Formula 1, characterized in that, at diffraction angle 2 θ X-ray powder diffraction pattern expressed in degrees 2Θ, having characteristic peaks at 11.288, 16.624, 18.251, 19.639, 21.140, 22.547, 23.439, 26.085, 26.497 and 27.050, 。 2. The A-type crystal according to claim 1, characterized in that, at diffraction angle 2 θ X-ray powder diffraction pattern expressed in degrees 2Θ, having characteristic peaks at 11.288, 16.624, 17.312, 18.251, 19.639, 20.086, 21.140, 22.547, 23.439, 24.234, 26.085, 26.497, and 27.

050.

3. The A-type crystal according to claim 1, characterized in that, at a diffraction angle 2 θ The X-ray powder diffraction pattern expressed in degrees is shown in Figure 1.

4. A method for preparing the A-type crystal as described in any one of claims 1-3, wherein the method is selected from: Method 1: Add the compound shown in Formula 1 to solvent (I) and slurry it to crystallize; the solvent (I) is selected from one of alcohol solvents, ketone solvents, ester solvents, ether solvents, hydrocarbon solvents, nitrile solvents, water, and nitromethane; The alcohol solvent is selected from methanol, ethanol, isopropanol, n-propanol, benzyl alcohol, 1,2-propanediol, and isoamyl alcohol; The ketone solvent is selected from acetone and methyl isobutyl ketone; The ester solvent is selected from isopropyl acetate; The ether solvent is selected from methyl tert-butyl ether, propylene glycol methyl ether, isopropyl ether, and tetrahydrofuran; The hydrocarbon solvent is selected from n-heptane, cyclohexane, toluene, and p-xylene; The nitrile solvent is selected from acetonitrile; Method 2: Dissolve the compound of Formula 1 in solvent (II) by heating, and stir at room temperature or cooling to allow crystallization; wherein the solvent (II) is selected from one of acetonitrile, acetone, nitromethane, benzyl alcohol, N,N-dimethylacetamide, dimethyl sulfoxide, N-methylpyrrolidone, acetonitrile / methanol; Method 3: Dissolve the compound of Formula 1 in dimethyl sulfoxide, add one of the following solvents: water, methanol, ethanol, isopropanol, isopropyl ether, or ethyl acetate, and stir to induce crystallization; or dissolve the compound of Formula 1 in dichloromethane, add one of the following solvents: methanol, ethanol, isopropanol, isopropyl ether, methyl tert-butyl ether, acetone, methyl isobutyl ketone, ethyl acetate, or toluene, and stir to induce crystallization.

5. A B-crystal form of the compound shown in Formula 1, characterized in that, at diffraction angle 2 θ X-ray powder diffraction pattern expressed in degrees 2Θ, having characteristic peaks at 6.454, 7.100, 11.249, 18.241, 19.637, 20.092, 20.407, 22.580, 26.107 and 26.550, 。 6. The B-type crystal according to claim 5, characterized in that, at diffraction angle 2 θ X-ray powder diffraction pattern expressed in degrees 2Θ, having characteristic peaks at 6.454, 7.100, 10.448, 11.249, 16.595, 18.241, 19.637, 20.092, 20.407, 22.580, 23.470, 26.107, and 26.

550.

7. The B-type crystal according to claim 5, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 2.

8. A method for preparing the B crystal form as described in any one of claims 5-7, the method comprising: The compound of Formula 1 was dissolved in solvent (V), and solvent (VI) was added and stirred to induce crystallization; the solvent (V) was selected from dimethyl sulfoxide; the solvent (VI) was selected from methyl tert-butyl ether.

9. A C-crystal form of the compound shown in Formula 1, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed as an angle, shows characteristic peaks at 6.484, 7.056, 7.907, 11.060, 12.702, 14.223, 15.973, 17.526, 20.342, 21.460, 23.090, 26.398, and 27.

500. 。 10. The C-type according to claim 9, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 3.

11. A method for preparing the C-type crystal as described in any one of claims 9-10, wherein the method is selected from: Method 1: Dissolve the compound of Formula 1 in solvent (VII), and add solvent (VIII) to precipitate crystals; wherein solvent (VII) is selected from dichloromethane; wherein solvent (VIII) is selected from tetrahydrofuran; Method 2: Dissolve the compound of Formula 1 in solvent (IX) and stir to crystallize; the solvent (IX) is selected from chloroform.

12. A D-crystal form of the compound shown in Formula 1, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed as an angle, shows characteristic peaks at 6.504, 7.051, 7.896, 11.086, 13.560, 14.194, 15.876, 17.504, 18.599, 20.238, 22.244, and 23.

963. 。 13. The D-type according to claim 12, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 6.504, 7.051, 7.896, 11.086, 13.560, 14.194, 15.876, 17.504, 18.599, 20.238, 20.633, 22.244, 23.963, and 26.

487.

14. The D-type according to claim 12, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 4.

15. A method for preparing the D-type crystal as described in any one of claims 12-14, the method comprising: The compound of Formula 1 was dissolved in solvent (X) and stirred to crystallize; the solvent (X) was selected from N,N-dimethylformamide.

16. An E-crystal form of the compound shown in Formula 1, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed as an angle, shows characteristic peaks at 6.490, 7.069, 8.034, 10.968, 14.194, 16.199, 18.656, 20.486, 21.539, 22.360, 22.949, 23.979, 24.676, and 26.

411. 。 17. The E-crystal form according to claim 16, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 5.

18. A method for preparing the E-type as described in any one of claims 16-17, the method comprising: The compound shown in Formula 1 was dissolved in solvent (XI) and stirred to crystallize; the solvent (XI) was selected from 1,2-dichloroethane.

19. An F-crystal form of the compound shown in Formula 1, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed as an angle, shows characteristic peaks at 7.327, 9.387, 13.968, 15.793, 20.198, and 21.

813. 。 20. The F-type according to claim 19, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 7.327, 8.690, 9.387, 9.653, 12.583, 13.968, 15.793, 17.270, 20.198, 20.810, 21.813, 22.186, and 25.

444.

21. The F-crystal form according to claim 19, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 7.327, 8.690, 8.970, 9.387, 9.653, 12.583, 13.968, 15.793, 16.429, 17.270, 17.975, 20.198, 20.810, 21.813, 22.186, 23.410, 24.438, 25.444, and 27.

356.

22. The F-crystal form according to claim 19, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 6.

23. A method for preparing the F-type crystal as described in any one of claims 19-22, the method comprising: The compound shown in Formula 1 was dissolved in solvent (XII) and stirred to crystallize; the solvent (XII) was selected from ethyl acetate.

24. An H-crystal form of the compound shown in Formula 1, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed as an angle, shows characteristic peaks at 5.271, 10.587, 13.230, 16.017, 21.498, 22.753, and 26.

938. 。 25. The H-crystal form according to claim 24, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 5.271, 10.587, 11.892, 13.230, 16.017, 16.625, 19.110, 21.498, 22.753, 24.080, 26.938, and 27.

536.

26. The H-crystal form according to claim 24, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 5.271, 10.587, 11.892, 13.230, 16.017, 16.625, 19.110, 21.498, 22.753, 24.080, 25.459, 26.175, 26.938, 27.536, 31.694, and 38.

116.

27. The H-crystal form according to claim 24, characterized in that, With diffraction angle 2 θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 8.

28. A method for preparing the H-type crystal as described in any one of claims 24-27, the method comprising: Step a. Dissolve the compound shown in Formula 1 in an aqueous sodium hydroxide solution and stir; and Step b. Add hydrochloric acid aqueous solution to the solid obtained in step a, and slurry and crystallize.

29. The crystal form according to any one of claims 1-3, 5-7, 9-10, 12-14, 16-17, 19-22, 24-27, wherein the 2 θ The angular error range is ±0.

20.

30. A pharmaceutical composition comprising the crystal form as described in any one of claims 1-3, 5-7, 9-10, 12-14, 16-17, 19-22, 24-27 and optionally a pharmaceutically acceptable excipient.

31. A method for preparing a pharmaceutical composition, comprising the step of mixing the crystal form according to any one of claims 1-3, 5-7, 9-10, 12-14, 16-17, 19-22, 24-27 with a pharmaceutically acceptable excipient.

32. Use of the crystal form or the composition of claim 30 as described in any one of claims 1-3, 5-7, 9-10, 12-14, 16-17, 19-22, 24-27 in the preparation of a medicament for the prevention and / or treatment of KAT6-mediated cancer.

33. Use of the crystal form or the composition of claim 30 as described in any one of claims 1-3, 5-7, 9-10, 12-14, 16-17, 19-22, 24-27 in the preparation of a KAT6 inhibitor.

34. Use of the crystal form or the composition of claim 30 as described in any one of claims 1-3, 5-7, 9-10, 12-14, 16-17, 19-22, 24-27 in the preparation of a medicament for the prevention and / or treatment of breast cancer.

35. Use of the crystal form as described in any one of claims 1-3, 5-7, 9-10, 12-14, 16-17, 19-22, 24-27 or the composition of claim 30 in the preparation of a medicament for the prevention and / or treatment of osteosarcoma.

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