Crystalline form of 3, 4-dihydro-2H-chromene [8, 7-d] isoxazole derivative and preparation method of 3, 4-dihydro-2H-chromene [8, 7-d] isoxazole derivative

Abstract

The invention relates to a crystalline form of a 3, 4-dihydro-2H-chromene [8, 7-d] isoxazole derivative and a preparation method of the 3, 4-dihydro-2H-chromene [8, 7-d] isoxazole derivative. The invention specifically provides a novel crystal form of a compound shown in a formula 1 and a preparation method of the novel crystal form. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to a crystalline form of a 3,4-dihydro-2H-chromeno[8,7-d]isoxazole derivative and a preparation method thereof. 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] The crystal form of the active pharmaceutical ingredient often affects the chemical stability of the drug. Different crystallization conditions and storage conditions may lead to changes in the crystal structure of the compound, and sometimes other forms of crystals may be produced. Generally speaking, amorphous drug products do not have a regular crystal structure and often have other defects, such as poor product stability, fine crystallization, difficult filtration, easy agglomeration, poor fluidity, etc. The polymorphic form of the drug has different requirements for product storage, production and scale-up. Therefore, it is necessary to conduct in-depth research on the crystal form of the aforementioned compound and improve the various properties of the aforementioned compound. Summary of the invention

[0007] The present disclosure provides a new crystal form of a compound represented by Formula 1, which has good stability and can be better applied in clinical practice.

[0008]

[0009] The crystalline form A of the compound represented by Formula 1 provided in the present disclosure has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, and has characteristic peaks at 7.472, 11.581, 20.815, 22.761 and 27.433.

[0010] In some embodiments, Form A of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 7.472, 11.581, 15.758, 16.732, 17.890, 20.815, 21.544, 22.761, 24.407 and 27.433.

[0011] In some embodiments, Form A of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 7.472, 11.329, 11.581, 15.758, 16.732, 17.890, 18.336, 20.815, 21.544, 22.761, 24.407 and 27.433.

[0012] In some embodiments, the crystalline form A of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 1 shown.

[0013] The present disclosure also provides a method for preparing the crystalline form A of the compound represented by Formula 1, the method being selected from:

[0014] Method 1: adding the compound represented by formula 1 into a solvent (I), and beating and crystallizing; the solvent (I) is selected from one or more of methanol, ethanol, and isopropanol;

[0015] Method 2: dissolving the compound of formula 1 in a solvent (II), evaporating the solvent to crystallize; the solvent (II) is selected from butanone.

[0016] The present disclosure also provides a crystalline form B of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 7.112, 11.559, 16.318, 20.040 and 21.212.

[0017] In some embodiments, the crystalline form B of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 7.112, 8.191, 11.559, 14.169, 16.318, 20.040, 21.212 and 24.504.

[0018] In some embodiments, the crystalline form B of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 2 shown.

[0019] The present disclosure also provides a method for preparing Form B, the method comprising:

[0020] Method 1: dissolving the compound of formula 1 in acetone, evaporating the solvent and crystallizing;

[0021] Method 2: Add the compound of formula 1 into acetone or a 5% water / acetone mixed solvent, and slurry and crystallize.

[0022] The present disclosure also provides a crystalline form C of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 6.939, 7.080, 11.499, 19.997 and 27.856.

[0023] In some embodiments, the crystalline form C of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 6.939, 7.080, 11.499, 16.064, 19.997, 24.321, 27.856 and 27.937.

[0024] In some embodiments, the crystalline form C of the compound represented by Formula 1 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 Form C, comprising adding the compound of Formula 1 to a solvent (III), and slurrying and crystallizing; the solvent (III) is selected from one or more of methyl sulfoxide, N,N-dimethylformamide, and N-methylpyrrolidone.

[0026] The present disclosure also provides a crystalline form D of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 5.348, 7.454, 14.959, 17.206 and 22.538.

[0027] In some embodiments, the crystalline form D of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 5.348, 7.454, 10.080, 14.959, 16.282, 17.206, 22.538 and 30.223.

[0028] In some embodiments, the crystalline form D of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 5.348, 7.454, 10.080, 14.959, 16.282, 17.206, 22.538, 30.223, 33.005 and 34.854.

[0029] In some embodiments, the crystalline form D of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 4 shown.

[0030] The present disclosure also provides a method for preparing the crystal form D, which comprises dissolving the compound of formula 1 in a solvent (IV), evaporating the solvent to crystallize; the solvent (IV) is selected from ethyl acetate.

[0031] The present disclosure also provides a crystalline form E of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 5.423, 6.977, 10.146 and 11.437.

[0032] In some embodiments, the crystalline form E of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 5.423, 6.977, 10.146, 11.437, 16.358 and 28.237.

[0033] In some embodiments, the crystalline form E of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 5 shown.

[0034] The present disclosure also provides a method for preparing the crystal form E, comprising dissolving the compound represented by Formula 1 in 10% water / acetone, and evaporating the solvent to crystallize.

[0035] The present disclosure also provides a crystalline form F of the compound represented by Formula 1, and an X-ray powder diffraction pattern represented by a diffraction angle 2θ has characteristic peaks at 6.986, 11.366, 13.990, 16.963 and 20.211.

[0036] In some embodiments, the crystalline form F of the compound of Formula 1 has characteristic peaks at 6.986, 7.165, 11.366, 13.990, 16.963, 18.482 and 20.211 in an X-ray powder diffraction pattern expressed as a diffraction angle 2θ.

[0037] In some embodiments, Form F of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ, with characteristic peaks at 6.986, 7.165, 11.366, 13.990, 16.963, 18.482, 20.211, 21.425, 22.157 and 25.259.

[0038] In some embodiments, the crystalline form F of the compound represented by Formula 1 has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in Figure 6 shown.

[0039] The present disclosure also provides a method for preparing the crystal form F, which comprises dissolving the compound represented by Formula 1 in dichloromethane, and evaporating the solvent to crystallize.

[0040] The present disclosure also provides a pharmaceutical composition, which contains the aforementioned crystalline form A, crystalline form B, crystalline form C, crystalline form D, crystalline form E, crystalline form F, and a pharmaceutical excipient selected from pharmaceutically acceptable excipients.

[0041] The present disclosure also provides a pharmaceutical composition, which is prepared from the aforementioned crystalline form A, crystalline form B, crystalline form C, crystalline form D, crystalline form E, crystalline form F, and optional pharmaceutically acceptable excipients.

[0042] The present disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned crystalline form A, crystalline form B, crystalline form C, crystalline form D, crystalline form E or crystalline form F with a pharmaceutically acceptable excipient.

[0043] The present disclosure also provides the use of the aforementioned crystalline form A, crystalline form B, crystalline form C, crystalline form D, crystalline form E or crystalline form F or the aforementioned composition in the preparation of a composition for preventing and / or treating cancer.

[0044] 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.

[0045] 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.

[0046] 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%.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] 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.

[0051] 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,

[0052] Deliquesce: Absorbs enough water to form a liquid;

[0053] Highly hygroscopic: weight gain due to moisture absorption is not less than 15%;

[0054] Hygroscopic: weight gain due to moisture absorption is less than 15% but not less than 2%;

[0055] Slightly hygroscopic: weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0056] No or almost no hygroscopicity: weight gain due to moisture is less than 0.2%.

[0057] 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

[0058] Figure 1 This is the XRPD spectrum of Compound 1 Form A.

[0059] Figure 2 This is the XRPD spectrum of Compound 1 Form B.

[0060] Figure 3 This is the XRPD spectrum of Compound 1 Form C.

[0061] Figure 4 This is the XRPD spectrum of Compound 1 Form D.

[0062] Figure 5 This is the XRPD spectrum of Compound 1 Form E.

[0063] Figure 6This is the XRPD spectrum of Compound 1 Form F.

[0064] Figure 7 This is the XRPD spectrum of the amorphous form of compound 1. DETAILED DESCRIPTION

[0065] 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.

[0066] Test conditions of the instruments used in the experiment:

[0067] 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.

[0068] 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).

[0069] High performance liquid chromatography (HPLC) analysis was performed using Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489.

[0070] High performance liquid chromatography was performed using Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson-281 preparative chromatographs.

[0071] Silica gel column chromatography generally uses Yantai Huanghai Silica Gel 200-300 mesh silica gel as the carrier.

[0072] 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°.

[0073] 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.

[0074] 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.

[0075] 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.

[0076] The known starting materials disclosed herein 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.

[0077] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0078] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1L.

[0079] 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.

[0080] Example 1 Preparation of the compound of formula 1

[0081] N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide 1

[0082] Refer to the preparation method of Example 3 of WO2023016484A1

[0083]

[0084] first step

[0085] 4-Bromo-2-((2,4-dimethoxybenzyl)oxy)-6-fluorobenzonitrile 1b

[0086] 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.

[0087] Step 2

[0088] 4-Bromo-2-fluoro-6-hydroxybenzonitrile 1c

[0089] 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%).

[0090] Step 3

[0091] 2-(Allyloxy)-4-bromo-6-fluorobenzonitrile 1d

[0092] 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%). 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).

[0093] Step 4

[0094] 3-Allyl-4-bromo-6-fluoro-2-hydroxybenzonitrile 1e

[0095] 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).

[0096] Step 5

[0097] 4-Bromo-6-fluoro-2-hydroxy-3-(3-hydroxypropyl)benzonitrile 1f

[0098] 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).

[0099] Step 6

[0100] 5-Bromo-7-fluorochromene-8-carbonitrile 1g

[0101] 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].

[0102] Step 7

[0103] 8-Cyano-7-fluorochromene-5-carboxylic acid methyl ester 1h

[0104] 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].

[0105] Step 8

[0106] 7-Fluoro-5-(hydroxymethyl)chromane-8-carbonitrile 1i

[0107] 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].

[0108] Step 9

[0109] 5-((1H-pyrazol-1-yl)methyl)-7-fluorochromene-8-carbonitrile 1k

[0110] 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].

[0111] Step 10

[0112] 5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-amine

[0113] 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%). 1 H 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].

[0114] Step 11

[0115] N-(5-((1H-pyrazol-1-yl)methyl)-3,4-dihydro-2H-chromeno[8,7-d]isoxazol-9-yl)-2-methoxybenzenesulfonamide 1

[0116] 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 C185μm 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 follows: Figure 7 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).

[0117] Example 2 Preparation of Crystalline Form A of the Compound Represented by Formula 1

[0118] Weigh about 20 mg of the compound of formula 1, dissolve it in 4 mL of butanone, filter it through a 0.45 μm filter membrane after it becomes clear, and slowly evaporate the solvent to obtain a solid. The product is defined as Form A by X-ray powder diffraction detection, and the XRPD spectrum is as follows: Figure 1 , and the positions of its characteristic peaks are shown in Table 1. The DSC spectrum shows that the peak value of the endothermic peak is 225.86℃. The TGA spectrum shows that the weight loss is 0.03% at 40℃-200℃. The DVS test shows that under normal storage conditions (i.e. 25℃, 60%RH), the sample has a moisture absorption weight gain of about 0.15%; under accelerated experimental conditions (i.e. 70%RH), the moisture absorption weight gain is about 0.19%; under extreme conditions (i.e. 90%RH), the moisture absorption weight gain is about 0.49%. After the DVS test, the crystal form was retested and the crystal form did not change.

[0119] Table 1

[0120]

[0121]

[0122]

[0123] Example 3 Preparation of Crystalline Form A of the Compound Represented by Formula 1

[0124] About 100 mg of the compound of formula 1 was weighed, 1 mL of solvent was added, the solvent was shown in Table 2 below, slurried at room temperature for 3 days, centrifuged and vacuum dried to obtain a solid. The product was crystal form A according to X-ray powder diffraction.

[0125] Table 2

[0126] Solvents Crystal form Methanol Crystalline Form A Ethanol Crystalline Form A Isopropyl alcohol Crystalline Form A

[0127] Example 4 Preparation of Crystalline Form B of the Compound Represented by Formula 1

[0128] Weigh about 20 mg of the compound shown in Formula 1, dissolve it in 2 mL of acetone, filter it through a 0.45 μm filter membrane, and slowly evaporate the solvent to obtain the title product. The product is defined as free form B by X-ray powder diffraction detection. The XRPD spectrum is as follows: Figure 2 The characteristic peak positions are shown in Table 3.

[0129] Table 3

[0130]

[0131]

[0132] Example 5 Preparation of Crystalline Form B of the Compound Represented by Formula 1

[0133] About 100 mg of the compound of Formula 1 was weighed, 1 mL of acetone was added, and the mixture was slurried at room temperature for 3 days. After centrifugation, the solid was vacuum dried to obtain a solid. The product was found to be Form B by X-ray powder diffraction.

[0134] Example 6 Preparation of Crystalline Form C of the Compound Represented by Formula 1

[0135] About 100 mg of the compound of Formula 1 was weighed, 1 mL of solvent was added, and the solvent was shown in Table 4 below. The mixture was slurried at room temperature for 3 days. After centrifugation, the solid was vacuum dried to obtain a solid. The product was defined as Form C by X-ray powder diffraction detection. The XRPD spectrum is shown in Figure 3 The characteristic peak positions are shown in Table 5.

[0136] Table 4

[0137] Solvents Crystal form Dimethyl sulfoxide Crystalline Form C N-Methylpyrrolidone Crystalline Form C N,N-Dimethylformamide Crystalline Form C

[0138] Table 5

[0139]

[0140]

[0141] Example 7 Preparation of Crystalline Form D of the Compound Represented by Formula 1

[0142] Weigh about 10 mg of the compound shown in Formula 1, add 4 mL of ethyl acetate to dissolve, filter through a 0.45 μm filter membrane after the solution becomes clear, and slowly evaporate the solvent to obtain a solid. The product is defined as Form D by X-ray powder diffraction detection, and the XRPD spectrum is as follows Figure 4 , and the characteristic peak positions are shown in Table 6.

[0143] Table 6

[0144]

[0145]

[0146] Example 8 Preparation of Crystalline Form E of the Compound Represented by Formula 1

[0147] Weigh about 10 mg of the compound shown in Formula 1, dissolve it in 2 mL of acetone containing 10% water, filter it through a 0.45 μm filter membrane after dissolving it, and slowly evaporate the solvent to obtain a solid. The product is defined as Form E by X-ray powder diffraction detection, and the XRPD spectrum is as follows Figure 5 , and the characteristic peak positions are shown in Table 7.

[0148] Table 7

[0149]

[0150]

[0151] Example 9 Preparation of Crystalline Form F of the Compound Represented by Formula 1

[0152] Weigh about 10 mg of the compound shown in Formula 1, dissolve it in 1 mL of dichloromethane, filter it through a 0.45 μm filter membrane, and slowly evaporate the solvent to obtain a solid. The product is defined as Form F by X-ray powder diffraction detection, and the XRPD spectrum is as follows: Figure 6 , and the characteristic peak positions are shown in Table 8.

[0153] Table 8

[0154]

[0155]

[0156] Example 10 Study on Hygroscopicity of Crystalline Form

[0157] Surface Measurement Systems intrinsic PLUS DVS was used to examine the humidity range of 0% to 95% at 25°C, with a step of 10% and a TMAX of 120min for one cycle.

[0158] Table 9

[0159] Crystal form 0.0%RH~80.0%RH Crystal form A 0.26％ constant

[0160] Example 11: Study on the stability of factors affecting crystal form

[0161] The free crystal form A and the sodium salt crystal form I were laid out in an open manner, and the stability of the samples was investigated under light (4500 Lux), high temperature (40° C., 60° C.), and high humidity (75% RH, 92.5% RH) conditions, respectively. The sampling period was 30 days.

[0162] Table 10

[0163]

[0164] 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), the physical and chemical stability of Form A are good.

[0165] Example 12: Long-term / accelerated stability of crystal forms

[0166] The free state samples were sealed in aluminum foil bags and placed at 40°C / 75% RH to investigate stability. The results are shown below.

[0167] Table 11

[0168]

[0169] Conclusion: Long-term accelerated experiments show that under 40℃ / 75%RH conditions for 1 month, Form A has good physical and chemical stability.

Claims

1. A crystalline form A 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 7.472, 11.581, 20.815, 22.761 and 27.433, preferably at 7.472, 11.581, 15.758, 16.732, 17.890, 20.815, 21.544, 22.761, 24.407 and 27.433, more preferably at 7.472, 11.329, 11.581, 15.758, 16.732, 17.890, 18.336, 20.815, 21.544, 22.761, 24.407 and 27.433, 2. The crystalline form A according to claim 1, characterized in that The X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ is shown in FIG1 .

3. A method for preparing the crystalline form A according to claim 1 or 2, wherein the method is selected from: Method 1: adding the compound represented by formula 1 into a solvent (I), and beating and crystallizing; the solvent (I) is selected from one or more of methanol, ethanol, and isopropanol; Method 2: dissolving the compound of formula 1 in a solvent (II), evaporating the solvent to crystallize; the solvent (II) is selected from butanone.

4. According to the crystal form described in claim 1-2, the 2θ angle error range is ±0.

20.

5. A pharmaceutical composition comprising the crystal form according to claims 1-2 and optionally a pharmaceutically acceptable excipient.

6. A method for preparing a pharmaceutical composition, comprising the step of mixing the crystal form according to claims 1-2 and a pharmaceutically acceptable excipient.

7. Use of the crystalline form according to claims 1-2, or the composition according to claim 5, in the preparation of a medicament for preventing and / or treating cancer.

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