Medicinal salt of 3, 4-dihydro-2H-chromene [8, 7-d] isoxazole derivative and crystal form thereof

Abstract

The present disclosure relates to a pharmaceutically acceptable salt of a 3, 4-dihydro-2H-chromene [8, 7-d] isoxazole derivative, and a crystalline form thereof. Specifically, the invention provides a pharmaceutically acceptable salt of N-(5-((1H-pyrazol-1-yl) methyl)-3, 4-dihydro-2H-chromene [8, 7-d] isoxazol-9-yl)-2-methoxybenzenesulfonamide, a crystal form and a preparation method thereof, and the corresponding salt has good stability and can be better used for clinical treatment.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to a usable salt, a crystal form and a preparation method of a 3,4-dihydro-2H-chromeno[8,7-d]isoxazole derivative. Background Art

[0002] 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 in protein substrates. Acetylation of lysine can affect the function of proteins, thereby playing an important regulatory role in chromosome structure, gene transcription regulation, DNA binding ability, enzyme activity and stability, protein interaction and intracellular localization. KATs are divided into several subfamilies, of which MYST (MOZ, YBF2 / SAS3, SAS2, TIP60) is the largest one, including KAT5 (TIP60), KAT6A (MOZ; MYST3), KAT6B (MORF; MYST4), KAT7 (HBO; MYST2) and KAT8 (MOF; MYST1). As the main members of the MYST family, KAT6A / B plays a vital role in development, maintenance of stem cells in the hematopoiesis and immune systems, and tumor development and drug resistance.

[0003] Published patent applications for KAT6 inhibitors include WO2016198507A1, WO2019243491A1, WO2019043139A1, WO2019108824A1, WO2020216701A1, WO2020002587A1, WO2020254946A1 and WO2020254989A1, etc.

[0004] WO2023016484 provides a KAT6 inhibitor, whose chemical name is N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide, having a structure shown in Formula 1,

[0005]

[0006] Salt formation can improve some undesirable physicochemical or biological properties of drugs. It is of great significance to develop salts with better physical and chemical properties or pharmaceutical properties than N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide. In view of the importance of solid drug crystal forms and their stability in clinical treatment, in-depth research on the polymorphic forms of pharmaceutically acceptable salts of the compound N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide is also of great significance for the development of drugs suitable for industrial production and with good biological activity. Summary of the invention

[0007] The present disclosure provides a pharmaceutically acceptable salt of the compound N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide shown in Formula I, wherein the pharmaceutically acceptable salt is selected from the group consisting of sodium salt, potassium salt, and calcium salt.

[0008]

[0009] The present disclosure also provides a method for preparing a pharmaceutically acceptable salt of the compound of formula 1, comprising the step of reacting the compound of formula 1, N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide, with a base selected from sodium hydroxide, potassium hydroxide, and calcium hydroxide.

[0010] Furthermore, in an optional embodiment, the method for preparing the aforementioned pharmaceutically acceptable salt further comprises the steps of crystallization, filtering, washing or drying.

[0011] The sodium salt I crystal form of the compound represented by Formula I provided in the present disclosure has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, and has characteristic peaks at 7.704, 10.316, 16.609, 22.917 and 24.772.

[0012] In some embodiments, the sodium salt I form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 6.840, 7.704, 9.462, 10.316, 11.007, 16.609, 19.681, 22.917 and 24.772.

[0013] In some embodiments, the sodium salt I form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 6.840, 7.704, 9.462, 10.316, 11.007, 11.818, 14.713, 15.398, 16.609, 19.681, 21.942, 22.917 and 24.772.

[0014] In some embodiments, the sodium salt I crystal form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 1 shown.

[0015] The present disclosure also provides a method for preparing a sodium salt crystalline form I of a compound of formula I, comprising adding a compound of formula 1 to acetone / water, adding sodium hydroxide, and slurrying and crystallizing.

[0016] The potassium salt I crystal form of the compound represented by formula I provided in the present disclosure has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, and has characteristic peaks at 6.450, 7.516, 11.088, 15.926, 21.929 and 25.383.

[0017] In some embodiments, the potassium salt I form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 6.450, 7.516, 9.156, 10.621, 11.088, 12.789, 14.014, 15.926, 21.929 and 25.383.

[0018] In some embodiments, the potassium salt I form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 6.450, 7.516, 9.156, 10.621, 11.088, 12.789, 14.014, 15.926, 19.376, 21.929, 22.650, 25.383 and 29.280.

[0019] In some embodiments, the potassium salt of the compound of Formula I, in Form I, has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 2 shown.

[0020] The present disclosure also provides a method for preparing a potassium salt crystalline form I of a compound of formula I, comprising adding a compound of formula 1 to acetone / water, adding potassium hydroxide, and slurrying and crystallizing.

[0021] The calcium salt I crystal form of the compound represented by formula I provided in the present disclosure has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, and has characteristic peaks at 7.072, 7.668, 11.314, 12.677 and 23.278.

[0022] In some embodiments, the calcium salt I crystal form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 7.072, 7.668, 10.393, 11.314, 12.677, 16.667, 23.278, 23.892 and 34.080.

[0023] In some embodiments, the calcium salt I crystal form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 7.072, 7.668, 10.393, 8.786, 8.949, 10.393, 11.314, 12.677, 16.667, 17.736, 18.028, 23.278, 23.892 and 34.080.

[0024] In some embodiments, the calcium salt of the compound of Formula I, in crystal form I, has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 3 shown.

[0025] The present disclosure also provides a method for preparing the calcium salt I crystal form of the compound shown in formula I, comprising adding the compound of formula 1 to acetone / water, adding calcium hydroxide, and slurrying and crystallizing.

[0026] The present disclosure also provides a pharmaceutical composition, which contains the aforementioned sodium salt I crystal form, potassium salt I crystal form, calcium salt I crystal form, and pharmaceutical excipients selected from pharmaceutically acceptable excipients.

[0027] The present disclosure also provides a pharmaceutical composition, which is prepared from the aforementioned sodium salt I crystal form, potassium salt I crystal form, calcium salt I crystal form, and optional pharmaceutically acceptable excipients.

[0028] The present disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned sodium salt form I, potassium salt form I, and calcium salt form I with a pharmaceutically acceptable excipient.

[0029] The present disclosure also provides the use of the aforementioned sodium salt I crystal form, potassium salt I crystal form, calcium salt I crystal form or the aforementioned composition in the preparation of a method for preventing and / or treating cancer.

[0030] The use disclosed herein, wherein the cancer is 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 cancer, esophageal cancer, essential thrombocythemia, Ewing's tumor, testicular cancer, glioma, heavy chain disease, Hemangioblastoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, neuroblastoma, NUT midline carcinoma, glioma, bone cancer, nasopharyngeal carcinoma, oral cancer, thyroid cancer, pinealoma, polycythemia vera, retinoblastoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, synovioma, sweat gland carcinoma, Waldenstrom's macroglobulinemia and Wilms' tumor; preferably, the cancer is selected from breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, gastric cancer, colorectal cancer and lung cancer.

[0031] The "2θ or 2θ angle" mentioned in the present disclosure refers to the diffraction angle, θ is the Bragg angle, and the unit is ° or degree; the error range of each characteristic peak 2θ is ±0.20 (including the case where the number exceeding 1 decimal place is rounded off), 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.

[0032] The numerical values ​​in the present disclosure, such as the data of the measured and calculated content of the relevant substances, inevitably have a certain degree of error. Generally speaking, ±10% is within the reasonable error range. There is a certain degree of error variation depending on the context where it is used, and the error variation does not exceed ±10%, and can be ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2% or ±1%, preferably ±5%.

[0033] The starting material used in the crystal preparation method of the present invention can be a compound in any form, and specific forms include but are not limited to: amorphous, any crystal form, hydrate, solvate, etc.

[0034] The drying temperature in the present disclosure is generally 25° C.-100° C., preferably 40° C.-70° C., and the drying can be performed under normal pressure or reduced pressure.

[0035] The crystallization methods described in the present disclosure include room temperature crystallization, cooling crystallization, volatile solvent crystallization, adding seed crystals to induce crystallization, etc. The cooling temperature is selected from below 65°C, preferably -10°C to 60°C, and stirring can also be performed during the crystallization process.

[0036] The "differential scanning calorimetry or DSC" described in the present disclosure refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining a constant temperature of the sample to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

[0037] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "9103 Drug Hygroscopicity Guidelines" in the fourth volume of the 2015 edition of the Chinese Pharmacopoeia,

[0038] Deliquesce: Absorbs enough water to form a liquid;

[0039] Highly hygroscopic: weight gain due to moisture absorption is not less than 15%;

[0040] Hygroscopic: weight gain due to moisture absorption is less than 15% but not less than 2%;

[0041] Slightly hygroscopic: weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0042] No or almost no hygroscopicity: weight gain due to moisture is less than 0.2%.

[0043] The "excipients" described in the present disclosure include, but are not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent or emulsifier approved by the U.S. Food and Drug Administration for use by humans or livestock animals. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 This is the XRPD spectrum of Compound 1 sodium salt Form I.

[0045] Figure 2 This is the XRPD spectrum of compound 1 potassium salt Form I.

[0046] Figure 3 This is the XRPD spectrum of Compound 1 calcium salt Form I.

[0047] Figure 4 This is the XRPD spectrum of the amorphous form of compound 1. DETAILED DESCRIPTION

[0048] The present disclosure will be explained in more detail below in conjunction with 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 essence and scope of the present disclosure.

[0049] Test conditions of the instruments used in the experiment:

[0050] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10 -6 The unit of (ppm) is given. NMR measurements were performed using a Bruker AVANCE NEO 500M NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD) as the measurement solvent, and tetramethylsilane (TMS) as the internal standard.

[0051] MS was measured using Agilent 1200 / 1290DAD-6110 / 6120 Quadrupole MS LC / MS (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), Waters ACQuity UPLC-QD / SQD (manufacturer: waters, MS model: waters ACQuity Qda Detector / waters SQ Detector), and THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0052] High performance liquid chromatography (HPLC) analysis was performed using Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489.

[0053] High performance liquid chromatography was performed using Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson-281 preparative chromatographs.

[0054] Silica gel column chromatography generally uses Yantai Huanghai Silica Gel 200-300 mesh silica gel as the carrier.

[0055] XRPD is X-ray powder diffraction detection: the measurement is carried out using a BRUKER D8 X-ray diffractometer, and the specific collection information is: Cu anode (40kV, 40mA), Cu-Kα1 ray Kα2 rays Kβ rays Scanning mode: θ / 2θ, scanning range (2 ranges): 3°-50°.

[0056] DSC is differential scanning calorimetry: the measurement was performed using a PerkinElmer DSC 6000 differential scanning calorimeter, with a heating rate of 10°C / min, a specific temperature range referring to the corresponding spectrum (mostly 40-300°C), and a nitrogen purge rate of 20 mL / min.

[0057] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA2 thermogravimetric analyzer, with a heating rate of 10°C / min, the specific temperature range refers to the corresponding spectrum (mostly 40-350°C), and a nitrogen purge rate of 20 mL / min.

[0058] DVS is dynamic moisture adsorption: the detection adopts SMS DVS InsttianscPlus, at 25°C, the humidity changes from 0% to 95% to 0%, the step is 10% (the last step is 5%) (the specific range of humidity is subject to the corresponding spectrum, and the most commonly used methods are listed here), and the step length is 120min.

[0059] The known starting materials disclosed in the present invention can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, Darui Chemicals and other companies.

[0060] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0061] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1L.

[0062] The reaction progress in the embodiment is monitored by thin layer chromatography (TLC), the developing solvent used in the reaction, the eluent system of column chromatography used for purifying the compound and the developing solvent system of thin layer chromatography include: A: n-hexane / ethyl acetate system, B: dichloromethane / methanol system, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.

[0063] Example 1 Preparation of the compound of formula 1

[0064] N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide 1

[0065] Refer to the preparation method of Example 3 of WO2023016484A1

[0066]

[0067] first step

[0068] 4-Bromo-2-((2,4-dimethoxybenzyl)oxy)-6-fluorobenzonitrile 1b

[0069] 4-Bromo-2,6-difluorobenzonitrile 1a (22 g, 101 mmol) and 2,4-dimethoxybenzyl alcohol (18.5 g, 110 mmol) were dissolved in N,N-dimethylformamide (200 mL), and cesium carbonate (49 g, 150 mmol) was added. The reaction solution was stirred at 60 ° C for 16 hours. The reaction solution was cooled to room temperature, filtered under reduced pressure, and the filtrate was diluted with ethyl acetate (500 mL) and washed with saturated sodium chloride solution (30 mL × 5). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title product 1b (36.9 g, yield: 100%). The product was used directly in the next step without purification.

[0070] Step 2

[0071] 4-Bromo-2-fluoro-6-hydroxybenzonitrile 1c

[0072] Compound 1b (36.9 g, 100.7 mmol) was dissolved in dichloromethane (250 mL), cooled to 0°C, trifluoroacetic acid (39 g, 342 mmol) was added dropwise, and the reaction solution was warmed to room temperature and stirred for 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using eluent system A to obtain the title product 1c (9.7 g, yield: 44.5%).

[0073] Step 3

[0074] 2-(Allyloxy)-4-bromo-6-fluorobenzonitrile 1d

[0075] Compound 1c (10.7 g, 49.5 mmol) was dissolved in N,N-dimethylformamide (120 mL), the reaction solution was cooled to 0°C, cesium carbonate (24 g, 73.7 mmol) and allyl bromide (11.2 g, 92.6 mmol) were added, and the reaction solution was heated to room temperature and stirred for 4 hours. The reaction solution was filtered under reduced pressure, the filtrate was diluted with ethyl acetate (400 mL), washed with saturated sodium chloride solution (30 mL×3), the obtained organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system A to obtain the title product 1d (11.7 g, yield: 92%). 1H 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).

[0076] Step 4

[0077] 3-Allyl-4-bromo-6-fluoro-2-hydroxybenzonitrile 1e

[0078] Compound 1d (3.35 g, 13.1 mmol) was dissolved in 1,2-dichlorobenzene (80 mL), nitrogen was replaced three times, and the reaction solution was stirred at 180°C for 13 hours. The reaction solution was cooled to room temperature, and the residue was purified by silica gel column chromatography (wet loading) with eluent system A to obtain 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).

[0079] Step 5

[0080] 4-Bromo-6-fluoro-2-hydroxy-3-(3-hydroxypropyl)benzonitrile 1f

[0081] 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) was added dropwise at 0°C. The reaction solution was stirred for 2 hours under an ice bath. Under an ice bath, 3M sodium hydroxide aqueous solution (13 mL, 39 mmol) and 30% hydrogen peroxide (3.0 mL) were added successively, and stirred for 10 minutes after addition. The pH of the reaction solution was adjusted to 2 with 2M hydrochloric acid, and extracted with ethyl acetate (100 mL×2). The organic phase was collected, dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent system A to obtain 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).

[0082] Step 6

[0083] 5-Bromo-7-fluorochromene-8-carbonitrile 1g

[0084] Compound 1f (3.8 g, 13.9 mmol) was dissolved in anhydrous tetrahydrofuran (80 mL), the reaction solution was cooled to 0°C, triphenylphosphine (4.4 g, 16.8 mmol) and diisopropyl azodicarboxylate (3.4 g, 16.8 mmol) were added, and the reaction solution was 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 with eluent system A to obtain the title product 1g (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].

[0085] Step 7

[0086] 8-Cyano-7-fluorochromene-5-carboxylic acid methyl ester 1h

[0087] Compound 1g (2.6g, 10.2mmol) was dissolved in a mixed solvent of 40mL methanol and N,N-dimethylformamide (V:V=1:3), and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (800mg, 1.09mmol) and triethylamine (3.0g, 2.93mmol) were added in sequence. Carbon monoxide was replaced 3 times and stirred at 10bar and 90℃ for 16 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, diluted with ethyl acetate (150mL), washed with saturated sodium chloride solution (50mL×3), and the obtained 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 with eluent system A to obtain the title product 1h (2.1g, 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].

[0088] Step 8

[0089] 7-Fluoro-5-(hydroxymethyl)chromane-8-carbonitrile 1i

[0090] Compound 1h (2.1 g, 8.93 mmol) was dissolved in dry tetrahydrofuran (40 mL), nitrogen was replaced 3 times, the reaction solution was cooled to 0 ° C, and lithium borohydride (2M, 18 mmol, 9.0 mL) 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), washed with saturated sodium chloride solution (50 mL × 2), and the obtained 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 with eluent system A to obtain the title product 1i (1.84 g, yield: 99.5%). MS m / z (ESI): 207.9 [M+1].

[0091] Step 9

[0092] 5-((1H-pyrazol-1-yl)methyl)-7-fluorochromene-8-carbonitrile 1k

[0093] Compound 1i (1.8 g, 8.69 mmol) and 1-(methylsulfonyl)-1H-pyrazole 1j (1.5 g, 10.3 mmol, prepared by the method disclosed in "Scheme 8 Intermediate 13 on page 61 of the specification of patent application WO2020254946A1") were dissolved in acetonitrile (30 mL), cesium carbonate (4.2 g, 12.9 mmol) was added, and the reaction was carried out at 70 ° C for 1 hour. Filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography with eluent system A to obtain the title product 1k (1.9 g, yield: 85.0%). MS m / z (ESI): 258.0 [M+1].

[0094] Step 10

[0095] 5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-amine

[0096] Compound 1k (1.9 g, 7.39 mmol) and acetohydroxamic acid (1.7 g, 22.2 mmol) were dissolved in N,N-dimethylformamide (30 mL) and water (4.0 mL), and potassium carbonate (6.2 g, 44.9 mmol) was added. The reaction solution was stirred at 70°C for 24 hours. The reaction solution was cooled to room temperature, water (100 mL) was added, filtered, the filter cake was collected, and dried to obtain 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].

[0097] Step 11

[0098] N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide 1

[0099] Compound 1l (300 mg, 1.11 mmol) and 2-methoxybenzenesulfonyl chloride 1m (460 mg, 2.22 mmol) were dissolved in pyridine (6.0 mL) and nitrogen was replaced 3 times. The reaction solution was subjected to microwave reaction 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 (Sharpsil-T Prep C18 5μm 30×150mm; mobile phase: A-aqueous phase (10mM ammonium bicarbonate): B-acetonitrile = 20%-45% (15min), flow rate: 30mL / min) to obtain the title product 1 (100 mg, yield: 20.5%). The product was amorphous as detected by X-ray powder diffraction. The XRPD spectrum is as shown below: Figure 4 MS m / z (ESI): 440.8 [M+1]. 1 H NMR(500MHz,DMSO-d6)δ9.96(s,1H),7.81-7.78(m,2H),7.62(d,1H),7.51(d,1H),7.20(d,1H),7.09(d ,1H),6.45(s,1H),6.32(t,1H),5.41(s,2H),4.19(t,2H),3.81(s,3H),2.69(t,2H),2.01-1.96(m,2H).

[0100] Example 2 Preparation of the sodium salt of the compound represented by formula 1 in form I

[0101] Weigh about 200 mg of the compound shown in Formula 1, add it to 3 mL of 5% acetone solution, add 20 mg of solid sodium hydroxide, stir and slurry overnight, centrifuge and vacuum dry the solid to obtain a solid. The product is defined as sodium salt I crystal form by X-ray powder diffraction detection, and the XRPD spectrum is as follows Figure 1 The positions of its characteristic peaks are shown in Table 1.

[0102] Table 1

[0103]

[0104]

[0105] Example 3 Preparation of the potassium salt of the compound represented by formula 1 in form I

[0106] Weigh about 200 mg of the compound shown in formula 2, add it to 3 mL of 5% acetone solution, add 28 mg of potassium hydroxide solid, stir and slurry overnight, centrifuge and vacuum dry the solid to obtain a solid. The product is defined as potassium salt I crystal form by X-ray powder diffraction detection, and the XRPD spectrum is as follows Figure 2 The positions of its characteristic peaks are shown in Table 2.

[0107] Table 2

[0108]

[0109]

[0110]

[0111] Example 4 Preparation of the crystalline form I of the calcium salt of the compound represented by formula 1

[0112] Weigh about 200 mg of the compound shown in formula 1, add it to 3 mL of 5% acetone solution, add 44 mg of calcium hydroxide solid, stir and slurry overnight, centrifuge and vacuum dry the solid to obtain a solid. The product is defined as calcium salt I crystal form by X-ray powder diffraction detection, and the XRPD spectrum is as follows Figure 3 The characteristic peak positions are shown in Table 3.

[0113] Table 3

[0114]

[0115]

[0116] Example 5 Study on the stability of factors affecting crystal form

[0117] The sodium salt I crystal form of the compound of formula 1 was laid open and placed flat, and the stability of the sample was investigated under light (4500 Lux), high temperature (40° C., 60° C.), and high humidity (75% RH, 92.5% RH) conditions, and the sampling period was 30 days.

[0118] Table 4

[0119]

[0120] Conclusion: The influencing factor experiment shows that under the conditions of light, high temperature (40°C, 60°C), and high humidity (75% RH, 92.5% RH) for 30 days, the physical and chemical stability of sodium salt I crystal form are good.

[0121] Example 6 Long-term / accelerated stability of crystal forms

[0122] The sodium salt I crystal sample of the compound of Formula 1 was sealed in an aluminum foil bag and placed under 40° C. / 75% RH conditions to investigate stability. The results are shown below.

[0123] Table 5

[0124]

[0125] Conclusion: Long-term accelerated experiments show that the physical and chemical stability of sodium salt form I is good under 40℃ / 75%RH conditions for 1 month.

Claims

1. A pharmaceutically acceptable salt of N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide represented by formula 1, wherein the pharmaceutically acceptable salt is selected from the group consisting of sodium salt, potassium salt and calcium salt.

2. A sodium salt I crystal form of the compound represented by formula 1, characterized in that: The X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 7.704, 10.316, 16.609, 22.917 and 24.772, preferably has characteristic peaks at 6.840, 7.704, 9.462, 10.316, 11.007, 16.609, 19.681, 22.917 and 24.772, more preferably has characteristic peaks at 6.840, 7.704, 9.462, 10.316, 11.007, 11.818, 14.713, 15.398, 16.609, 19.681, 21.942, 22.917 and 24.

772.

3. The sodium salt I crystal form according to claim 2, characterized in that: The X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ is shown in FIG1 .

4. A method for preparing the sodium salt I crystal form as claimed in claim 2 or 3, comprising adding the compound of formula 1 to acetone / water, adding sodium hydroxide, and slurrying and crystallizing.

5. According to the crystal form according to any one of claims 2-3, the 2θ angle error range is ±0.

20.

6. A pharmaceutical composition comprising the crystal form according to any one of claims 2 to 3 and optionally a pharmaceutically acceptable excipient.

7. A method for preparing a pharmaceutical composition, comprising the step of mixing the crystal form according to any one of claims 2 to 3 and a pharmaceutically acceptable excipient.

8. Use of the crystal form according to any one of claims 2 to 3, or the pharmaceutical composition according to claim 6, in the preparation of a medicament for treating and / or preventing cancer.

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