Crystalline forms of 1H-pyrrolo[2,3-c]pyridine compounds and their manufacturing methods

Abstract

The present invention provides a crystalline form of 1H-pyrrolo[2,3-c]pyridine compound and a method for preparing the same, and further includes the use of said compound (II) and its crystalline form in the preparation of a medicament for treating related diseases. TIFF2025510077000021.tif3761

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims priority to application number CN202210307050.9, filed on March 25, 2022.

[0002] [Technical field] The present invention relates to crystalline forms of 1H-pyrrolo[2,3-c]pyridine compounds and methods for preparing the same, and further includes the use of said compounds and crystalline forms thereof in the preparation of therapeutic agents for related diseases. [Background technology]

[0003] Colony-stimulating factor 1 (CSF-1, also known as macrophage colony-stimulating factor or M-CSF) is an important growth factor that regulates the development of bone marrow progenitor cells, monocytes, macrophages, and their differentiated forms, osteoclasts and dendritic cells. To exert its biological effects, it must bind to its sole cell surface receptor, CSF-1R. CSF-1R, also known as c-FMS because it is encoded by the proto-oncogene c-FMS, is a receptor tyrosine kinase. Binding of CSF-1 and CSF-1R via their extracellular domains induces CSF-1R dimerization, which then triggers autophosphorylation of the intracellular CSF-1R kinase domain. Upon phosphorylation, CSF-1R serves as a docking site for several cytoplasmic signaling molecules, ultimately triggering a series of signaling cascades. For example, phosphorylation of the tyrosine residue at position 697 of CSF-1R can activate the MAPK signaling pathway, and phosphorylation of the tyrosine residue at position 721 can activate the PI3K and PLCγ signaling pathways, etc.

[0004] Colony-stimulating factor 1 receptor (CSF-1R) is a key target in the regulation of tumor-associated macrophages within the tumor microenvironment. Many tumor cells can secrete growth factors such as CSF-1 during their growth, which can recruit macrophages (tumor-associated macrophages, or TAMs) to the tumor area. Similar to tumor cells, macrophages can also secrete CSF-1, and their addition promotes the formation of a complex tumor microenvironment. This microenvironment helps tumor cells develop immune tolerance to their own immune system, thereby promoting tumor cell proliferation, invasion, and metastasis in the body. Blocking the CSF-1 / CSF1R pathway has been shown to significantly reduce macrophage infiltration into tumor sites, slow primary tumor growth, and reduce tumor metastasis. Therefore, inhibiting macrophage survival / activation by inhibiting CSF-1 / CSF1R signaling has become an important strategy for cancer immunotherapy.

[0005] Recent studies have demonstrated that CSF-1R inhibitors can be used in a variety of ways in the field of disease treatment. They can be used alone or in combination with various anti-cancer therapies, such as antiangiogenesis, adoptive T cell transfer, radiation therapy, chemotherapy, and immune checkpoint therapy. Many commercially available drugs, such as imatinib, dasatinib, and sunitinib, have inhibitory activity against CSF-1R. However, selective CSF-1R inhibitors are not yet commercially available. Pexidartinib (PLX-3397), developed by Plexxikon and acquired by Daiichi Sankyo, is a dual CSF-1R and c-Kit inhibitor and was approved by the FDA in August 2019 for the treatment of tenosynovial giant cell tumor (TGCT). [ka] Summary of the Invention

[0006] The present invention provides crystalline form A of compound of formula (I), which is characterized by the presence of characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 9.25±0.20°, and 14.45±0.20° in a powder X-ray diffraction pattern. [ka]

[0007] In some embodiments of the present invention, the crystalline form A has characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 7.98±0.20°, 9.25±0.20°, 14.45±0.20°, 16.02±0.20°, and 24.52±0.20° in a powder X-ray diffraction pattern.

[0008] In some embodiments of the present invention, the crystalline form A has characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 7.98±0.20°, 9.25±0.20°, 12.21±0.20°, 14.45±0.20°, 16.02±0.20°, 20.20±0.20°, and 24.52±0.20° in a powder X-ray diffraction pattern.

[0009] In some embodiments of the present invention, the crystalline form A has characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 7.98±0.20°, 9.25±0.20°, 10.08±0.20°, 12.21±0.20°, 14.45±0.20°, 15.13±0.20°, 16.02±0.20°, 20.20±0.20°, 21.35±0.20°, 21.96±0.20°, and 24.52±0.20° in a powder X-ray diffraction pattern.

[0010] In some embodiments of the present invention, the crystalline form A has characteristic diffraction peaks at 2θ angles of 6.12°, 7.98°, 9.25°, 10.08°, 12.21°, 14.45°, 15.13°, 16.02°, 18.34°, 20.20°, 21.35°, 21.96°, 23.41°, 24.52°, 25.43°, 27.39°, 28.56°, 29.03°, 29.96°, 31.75°, and 37.38° in a powder X-ray diffraction pattern.

[0011] In some embodiments of the present invention, the crystalline form A has an X-ray powder diffraction pattern of 6.12±0.20°, 9.25±0.20°, and / or 14.45±0.20°, and / or 7.98±0.20°, and / or 16.02±0.20°, and / or 24.52±0.20°, and / or 12.21±0.20°, and / or 20.20±0.20°, and / or 10.08±0.20°, and / or 15.13±0.20°, and / or 1 Characteristic diffraction peaks are present at 2θ angles of 8.34±0.20°, and / or 21.35±0.20°, and / or 21.96±0.20°, and / or 23.41±0.20°, and / or 25.43±0.20°, and / or 27.39±0.20°, and / or 28.56±0.20°, and / or 29.03±0.20°, and / or 29.96±0.20°, and / or 31.75±0.20°, and / or 37.38±0.20°. In some embodiments of the present invention, crystalline Form A above has an XRPD pattern substantially as shown in FIG.

[0012] In some embodiments of the present invention, analytical data for the XRPD pattern of crystalline form A above is shown in Table 1. [Table 1]

[0013] In some embodiments of the present invention, the differential scanning calorimetry curve for crystalline form A above has an endothermic peak with an onset at 200.9±3.0° C.

[0014] In some embodiments of the present invention, the DSC pattern of crystalline form A above is as shown in FIG.

[0015] The present invention further provides a compound of formula (II). [ka]

[0016] The present invention provides crystalline form B of compound of formula (II), which is characterized by the presence of characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, and 15.918±0.200° in a powder X-ray diffraction pattern. [ka]

[0017] In some embodiments of the present invention, the crystalline form B has characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, 13.019±0.200°, 15.918±0.200°, 19.697±0.200°, and 25.200±0.200° in a powder X-ray diffraction pattern.

[0018] In some embodiments of the present invention, the crystalline form B has characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, 7.941±0.200°, 11.218±0.200°, 13.019±0.200°, 15.918±0.200°, 19.697±0.200°, and 25.200±0.200° in a powder X-ray diffraction pattern.

[0019] In some embodiments of the present invention, the crystalline form B has characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, 7.941±0.200°, 11.218±0.200°, 13.019±0.200°, 14.701±0.200°, 15.918±0.200°, 17.338±0.200°, 19.697±0.200°, 22.540±0.200°, 25.200±0.200°, and 27.798±0.200° in a powder X-ray diffraction pattern.

[0020] In some embodiments of the present invention, the crystalline form B has an X-ray powder diffraction pattern of 4.579±0.200°, 6.898±0.200°, and / or 15.918±0.200°, and / or 13.019±0.200°, and / or 19.697±0.200°, and / or 25.200±0.200°, and / or 7.941±0.200°, and / or 11.218±0.200°, and / or 9.070±0.200°, and / or 9.423±0.200°, and / or 13.796±0.200°, and / or 14.701±0.200°, and / or 15.182±0.200°, and / or 17.338±0.200°, and / or 18.939±0.200°, and / or 19.461±0.200°, and / or 20.700±0.200°, and / or 21.602±0.200°, and / or 22.200±0.200°, and / or 22 .540±0.200°, and / or 22.818±0.200°, and / or 23.919±0.200°, and / or 24.461±0.200°, and / or 25.639±0.200°, and / or 26.563±0.200°, and / or 27.120±0.200°, and / or 27.798±0.200°, and / or 28.156±0.200°, and / or 28.595±0.200°, and / or 29.000±0 Characteristic diffraction peaks are present at 2θ angles of 0.200°, and / or 29.339±0.200°, and / or 30.739±0.200°, and / or 32.064±0.200°, and / or 32.355±0.200°, and / or 33.201±0.200°, and / or 34.156±0.200°, and / or 35.020±0.200°, and / or 36.063±0.200°, and / or 39.176±0.200°.

[0021] In some embodiments of the present invention, the crystalline form B has the following X-ray powder diffraction patterns: 4.579°, 6.898°, 7.941°, 9.070°, 9.423°, 11.218°, 13.019°, 13.796°, 14.701°, 15.182°, 15.918°, 17.338°, 18.939°, 19.461°, 19.697°, 20.700°, 21.602°, 22.200°, 22.54° Characteristic diffraction peaks are present at 2θ angles of 0°, 22.818°, 23.919°, 24.461°, 25.200°, 25.639°, 26.563°, 27.120°, 27.798°, 28.156°, 28.595°, 29.000°, 29.339°, 30.739°, 32.064°, 32.355°, 33.201°, 34.156°, 35.020°, 36.063°, and 39.176°.

[0022] In some embodiments of the present invention, crystalline form B above has an XRPD pattern substantially as shown in FIG.

[0023] In some embodiments of the present invention, analytical data for the XRPD pattern of crystalline form B above is shown in Table 2. [Table 2]

[0024] In some embodiments of the present invention, the differential scanning calorimetry curve for crystalline form B has an endothermic peak at 240.16±3.0°C and an exothermic peak at 282.04±3.0°C.

[0025] In some embodiments of the present invention, the DSC pattern of crystalline form B is shown in FIG.

[0026] In some embodiments of the present invention, the thermogravimetric analysis curve of the above crystalline form B shows a weight loss of 0.3643% at 200.0±3° C.

[0027] In some embodiments of the present invention, the TGA pattern of crystalline form B is as shown in FIG.

[0028] The present invention provides crystalline form C of compound of formula (III), which is characterized by the presence of characteristic diffraction peaks at 2θ angles of 5.35±0.20°, 8.66±0.20°, and 14.22±0.20° in a powder X-ray diffraction pattern. [ka]

[0029] In some embodiments of the present invention, the crystalline form C has characteristic diffraction peaks at 2θ angles of 5.35±0.20°, 5.91±0.20°, 8.66±0.20°, 13.12±0.20°, 14.22±0.20°, and 25.19±0.20° in a powder X-ray diffraction pattern.

[0030] In some embodiments of the present invention, the crystalline form C has characteristic diffraction peaks at 2θ angles of 5.35±0.20°, 5.91±0.20°, 8.66±0.20°, 13.12±0.20°, 14.22±0.20°, 17.61±0.20°, 25.19±0.20°, and 26.34±0.20° in a powder X-ray diffraction pattern.

[0031] In some embodiments of the present invention, the crystalline form C has an X-ray powder diffraction pattern of 5.35±0.20°, 5.91±0.20°, and / or 8.66±0.20°, and / or 13.12±0.20°, and / or 14.22±0.20°, and / or 17.61±0.20°, and / or 25.19±0.20°, and / or 26.34±0.20°, and / or Characteristic diffraction peaks are present at 2θ angles of 7.17±0.20°, and / or 10.63±0.20°, and / or 17.34±0.20°, and / or 21.22±0.20°, and / or 22.28±0.20°, and / or 29.39±0.20°, and / or 31.03±0.20°, and / or 32.10±0.20°, and / or 34.91±0.20°.

[0032] In some embodiments of the present invention, the crystalline form C has characteristic diffraction peaks at 2θ angles of 5.35°, 5.91°, 7.17°, 8.66°, 10.63°, 13.12°, 14.22°, 17.34°, 17.61°, 21.22°, 22.28°, 25.19°, 26.34°, 29.39°, 31.03°, 32.10°, and 34.91° in a powder X-ray diffraction pattern.

[0033] In some embodiments of the present invention, crystalline form C above has an XRPD pattern substantially as shown in FIG.

[0034] In some embodiments of the present invention, analytical data for the XRPD pattern of crystalline form C above is shown in Table 3. [Table 3]

[0035] In some forms of the present invention, the differential scanning calorimetry curve for crystalline Form C above has an endothermic peak with an onset at 223.2±3.0° C.

[0036] In some embodiments of the present invention, the DSC pattern of crystalline form C is as shown in FIG.

[0037] The present invention provides Step 1) adding a compound of formula (I) to a solvent to form a solution; Step 2) adding 12N concentrated hydrochloric acid to the solution, stirring at a certain temperature for a certain time, filtering, and vacuum drying the filter cake; the solvent is an alcohol-based solvent, The stirring temperature is 30 to 50°C, The method for preparing crystalline form B of compound of formula (II) is further provided, wherein the stirring time is 12 to 24 hours.

[0038] The present invention provides Step 1) adding a compound of formula (I) to a solvent to form a solution; Step 2) adding 12N concentrated hydrochloric acid to the solution, stirring at a certain temperature for a certain time, filtering, and vacuum drying the filter cake; the solvent is an alcohol-based solvent, The stirring temperature is 40 to 45°C. The present invention further provides a method for preparing crystalline form B of compound of formula (II), wherein the stirring time is 12 to 16 hours.

[0039] The present invention further provides the use of the compound of formula (II), crystalline form A, crystalline form B or crystalline form C, as described above, in the manufacture of a medicament for the treatment of tenosynovial giant cell tumor.

[0040] [Definitions and Explanations] Unless otherwise specified, the following terms and phrases used herein shall have the following meanings: Certain words or terms should not be considered uncertain or unclear in the absence of a specific definition and should be understood in their ordinary sense. When trade names are mentioned herein, it is intended to refer to the corresponding trade name or its active ingredient.

[0041] The intermediate compounds of the present invention can be prepared by various synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalents known to those skilled in the art. Preferred embodiments include, but are not limited to, embodiments of the present invention.

[0042] The chemical reactions of certain embodiments of the present invention are carried out in suitable solvents. The solvent must be suitable for the chemical transformations of the present invention and the required reagents and materials. To obtain the compounds of the present invention, those skilled in the art may need to modify or select synthetic steps or reaction procedures based on existing embodiments.

[0043] The structure of the compounds of the present invention can be confirmed by conventional methods known to those skilled in the art. When the present invention involves the absolute configuration of a compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, in single crystal X-ray diffraction (SXRD), a Bruker D8 venture diffractometer is used to collect diffraction intensity data from a cultured single crystal. The source is CuKα radiation, and the scan mode is φ / ω scan. After collecting the relevant data, the absolute configuration can be confirmed by further analyzing the crystal structure using a direct method (Shelxs97).

[0044] The present invention will be described in more detail below with reference to examples, but these examples are not intended to limit the present invention in any way.

[0045] All solvents used in this invention are commercially available and can be used without further purification.

[0046] The solvents used in the present invention are commercially available.

[0047] Compounds are named according to conventional naming principles in the art or using ChemDraw® software; commercially available compounds use supplier catalog names. [Effects of the Invention]

[0048] The crystalline form of the compound of the present invention is stable and slightly hygroscopic, and can significantly inhibit the proliferation of BCR-FMS-BaF3 cells.

[0049] X-ray powder diffractometer (XRPD) method of the present invention Crystalline Form A of Compound of Formula (I) and Crystalline Form C of Compound of Formula (III): Instrument name: PANalytical X'pert 3 Type X-ray diffractometer Test method: Approximately 10 mg of sample is spread evenly on a single crystal silicon sample plate. [Table 4]

[0050] Crystalline form B of compound of formula (II): Name of instrument: X-ray diffraction equipment Instrument model: DX-2700BH Equipment manufacturer: Dandong Haoyuan Instrument Co., Ltd.

[0051] Method parameters: Tube: Cu, k-Alphal (λ=1.54184Å) Tube voltage: 40 kV, tube current: 30 mA Divergence slit: 1mm Detector slit: 0.3 mm Anti-scatter slit: 1mm Scan range: 3~40deg Step angle: 0.02 deg Step size: 0.5 seconds Testing method: Place the sample on the sample plate, flatten the surface of the sample plate, and then place the sample plate in the X-ray diffraction device for testing.

[0052] Differential Scanning Calorimeter (DSC) Method of the Present Invention Crystalline Form A of Compound of Formula (I) and Crystalline Form C of Compound of Formula (III): Instrument model: TA2500 differential scanning calorimeter [Table 5]

[0053] Crystalline form B of compound of formula (II): Instrument model: Mettler Toledo DSC 1 Differential Scanning Calorimeter Test method: A sample (2.44 mg) is placed in a DSC high-pressure crucible, pressed and sealed, and then tested. The sample is heated from 40°C to 350°C at a heating rate of 10°C / min.

[0054] Thermogravimetric Analysis (TGA) Method of the Present Invention Instrument model: TGA2SF / 1100 Test method: A sample (2.745 mg) is placed in an alumina crucible and heated from 40°C to 500°C at a rate of 10°C / min.

[0055] Dynamic Vapor Sorption Analysis (DVS) Method of the Present Invention Instrument model: SMS DVS Intrinsic Dynamic Vapor Sorption Apparatus Test conditions: The sample (10 mg) is placed in the DVS sample plate and tested. The detailed DVS parameters are as follows: Temperature: 25℃ Equilibration: dm / dt=0.01% / min Drying: 25℃, 0% RH for 2 hours RH(%) test gradient: 5% RH RH(%) test gradient range: 0%-95%-0% RH

[0056] [Table 6] [Brief explanation of the drawings]

[0057] [Figure 1] 1 is an XRPD spectrum of Cu-Kα radiation of crystalline form A of compound of formula (I). [Figure 2] 1 is a DSC spectrum of crystalline form A of compound of formula (I). [Figure 3] 1 is an XRPD spectrum of Cu-Kα radiation of crystalline form B of compound of formula (II). [Figure 4] 1 is a DSC spectrum of crystalline form B of compound of formula (II). [Figure 5] 1 is a TGA spectrum of crystalline form B of compound of formula (II). [Figure 6] 1 is an XRPD spectrum of Cu-Kα radiation of crystalline form C of compound of formula (III). [Figure 7] 1 is a DSC spectrum of crystalline form C of compound of formula (III). [Figure 8] 1 is a DVS spectrum of crystalline form B of compound of formula (II). DETAILED DESCRIPTION OF THE INVENTION

[0058] In order to better understand the present invention, the present invention will be further described below with reference to specific examples, but the specific examples are not intended to limit the present invention.

[0059] Example 1: Preparation of Compound of Formula (I) [ka]

[0060] Step 1: Synthesis of compound 1-B 1-A (125 g, 819.24 mmol) was dissolved in methanol (2.5 L) and toluene (2.5 L), and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex (40 g, 48.98 mmol) and triethylamine (331.59 g, 3.28 mol) were added. The mixture was then stirred in an autoclave at an external temperature of 110 °C under a carbon monoxide (2 MPa) atmosphere for 40 hours. After the reaction was completed, the mixture was filtered through diatomaceous earth and concentrated under reduced pressure to remove the solvent, yielding a crude product. The crude product was purified by column chromatography (n-heptane:ethyl acetate = 1:1) to yield compound 1-B. MS m / z: 177.2 [M+H] +

[0061] Step 2: Synthesis of compound 1-C 1-B (249.9 g, 1.42 mol) was dissolved in N,N-dimethylformamide (2.5 L), and the reaction mixture was cooled to 0°C. N-iodosuccinimide (351.05 g, 1.56 mol) was added, and the reaction mixture was stirred at 0-5°C for 1 hour. The reaction mixture was slowly poured into water (9 L), stirred for 15 minutes, and then filtered under reduced pressure to obtain a filter cake. The filter cake was washed twice with 4 L of water. The filter cake was baked in an oven under reduced pressure, and then ethanol (800 mL) was added. The mixture was stirred for 1 hour and then filtered under reduced pressure to obtain product 1-C. MS m / z: 302.7 [M+H] +

[0062] Step 3: Synthesis of compound 1-D 1-C (200 g, 662.10 mmol) was dissolved in tetrahydrofuran (3 L). The reaction mixture was cooled to 0-5°C and sodium hydride (39.73 g, 993.15 mmol, 60% content) was slowly added. After the addition was complete, the mixture was stirred at 0-5°C for 0.5 h. 2-(chloromethoxy)ethyltrimethylsilane (139.09 g, 834.25 mmol) was then added. The reaction mixture was allowed to warm to 20°C and stirred for 1 h. Under a weak nitrogen flow, the reaction mixture was slowly poured into 3 L of water and quenched with stirring. Ethyl acetate (3 L) was added for extraction, and the organic phase was washed with water (3 L) and then with saturated brine (3 L). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to remove the solvent, yielding the crude product. The crude product was purified by column chromatography (n-heptane:tetrahydrofuran = 7:1 to 4:1) to obtain a crude product. To the obtained crude product, n-heptane (2 L) was added and stirred for 1 hour, and then suction filtered under reduced pressure to obtain product 1-D. MS m / z: 433.1 [M+H] +

[0063] Step 4: Synthesis of compound 1-F 1-E (2.5 kg, 15.77 mol) and cesium carbonate (10.28 kg, 31.54 mol) were dissolved in tetrahydrofuran (10 L), the system was cooled to 0-10°C, and cyclopropanol (1.14 kg, 19.71 mol) was added dropwise. After the addition was completed, the reaction solution was slowly heated to 25-30°C and stirred for 16 hours. The reaction solution was suction filtered under reduced pressure, and the resulting filter cake was rinsed twice with tetrahydrofuran (1 L). The resulting filtrate was combined with the rinsed filtrate. The resulting filtrate was slowly poured into water (36 L), stirred for 30 minutes, and then suction filtered under reduced pressure to obtain a filter cake. Water (5 L) was added to the resulting filter cake, stirred for 30 minutes, and then suction filtered under reduced pressure to obtain product 1-F. MS m / z: 180.8 [M+H] +

[0064] Step 5: Synthesis of compound 1-G 1-F (100 g, 555.06 mmol) was dissolved in ethanol (1 L), Pd / C (5 g, 555.06 mmol, 10% content) was added, and the reaction was stirred under a hydrogen atmosphere (30-40 psi) at 25-30°C for 16 hours. The reaction mixture was filtered through diatomaceous earth, and the solvent was removed by rotary evaporation under reduced pressure to give 1-G. MS m / z: 151.2 [M+H] +

[0065] Step 6: Synthesis of compound 1-H 1-G (154 g, 1.03 mol) was dissolved in 1,4-dioxane (2.25 L) and water (45 mL), and 2,5-dibromo-4-methylpyridine (308.77 g, 1.23 mol), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (29.67 g, 51.27 mmol), and potassium carbonate (354.31 g, 2.56 mol) were added thereto, and the system was purged with nitrogen three times. Tris(dibenzylideneacetone)dipalladium(0) (14.74 g, 25.64 mmol) was added thereto, and the system was purged with nitrogen three times. The reaction solution was stirred at 55 to 60°C under nitrogen protection for 16 hours. The reaction mixture was filtered through diatomaceous earth, and the filter cake was rinsed with 1,4-dioxane (500 mL × 3). The obtained filtrate (volume: approximately 6 L) was concentrated under reduced pressure to a volume of approximately 3.5 L, slowly poured into n-heptane (17.5 L), stirred for 0.5 hours, and then suction filtered under reduced pressure to obtain a filter cake. Water (3 L) was added to the filter cake, and the mixture was stirred for 20 minutes. The mixture was then suction filtered under reduced pressure to obtain 1-H. MS m / z: 319.9 [M+H] +

[0066] Step 7: Synthesis of Compound 1-I 1-H (2.18 kg, 6.80 mol) was dissolved in dichloromethane (17.5 L) and 4-dimethylaminopyridine (41.56 g, 340.21 mmol) was added. Di-tert-butyl dicarbonate (1.93 kg, 8.85 mol) was then added dropwise, and the reaction mixture was stirred at an external temperature of 30-40°C for 3 hours. The reaction mixture was washed with water (10 L), and the organic phase was washed with saturated brine (10 L). The organic phase was collected, dried over anhydrous sodium sulfate (1 kg), and concentrated under reduced pressure to obtain the crude product. n-Heptane (5 L) was added to the crude product, and the mixture was stirred at 0°C in an ice bath for 15 minutes. After stirring for another hour, the solid gradually precipitated. The mixture was then filtered with suction to obtain 1-I. MS m / z: 420.1 [M+H] +

[0067] Step 8: Synthesis of Compound 1-J 1-I (500.31 g, 1.19 mol), bis(pinacolato)diboron (392.81 g, 1.55 mol), dioxane (5 L), and potassium acetate (236.73 g, 2.38 mol) were added to a reaction vessel in this order, and the reaction system was purged with nitrogen three times. Thereafter, [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (43.71 g, 0.08 mol) was added, and the reaction system was purged with nitrogen three times. After that, the reaction system was protected with a nitrogen stream and heated to 70-80°C and stirred for 12-16 hours. The reaction mixture was filtered under reduced pressure through diatomaceous earth (200 g), the filter cake was rinsed with ethyl acetate (1 L × 2), the combined filtrates were concentrated, and the resulting crude product was purified by column chromatography (eluent: n-heptane:tetrahydrofuran = 10 / 1) to obtain compound 1-J. MS m / z: 490.3 [M+H+Na] +

[0068] Step 9: Synthesis of Compound 1-K 1-D (385.26 g, 0.89 mol), 1-J (466.95 g, 0.91 mol), and dioxane (3.85 L) were added sequentially to a reaction vessel and stirred. Potassium phosphate (385.81 g, 1.82 mol) was dissolved in water (385 mL) and added to the reaction vessel at room temperature (20-30 °C). The reaction system was purged with nitrogen three times. The reaction system was cooled to 0-5 °C, and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex (65.25 g, 0.09 mol) was added. The reaction system was purged with nitrogen three times. The reaction system was heated to 35-40 °C and stirred for 12-16 hours. Ethyl acetate (8 L) and water (8 L) were added to the reaction solution, stirred for 10 minutes, and then allowed to stand for separation. The aqueous phase was separated. The organic phase was washed with water (8 L) and saturated sodium chloride solution (8 L). The organic phase was collected, dried over anhydrous sodium sulfate (500 g), suction filtered under reduced pressure, and concentrated under reduced pressure to obtain a crude product. Ethyl acetate (7.7 L) and powdered activated carbon (193.38 g) were added to the crude product, and the mixture was refluxed and stirred at 75-80°C for 1 hour. The temperature was then lowered to 55-60°C, and the crude product was filtered under reduced pressure through diatomaceous earth (500 g). The filter cake was rinsed with ethyl acetate (15 L). The combined filtrate was concentrated under reduced pressure to approximately 2 L (temperature below 45°C). Ethyl acetate (0.3 L) and n-heptane (3 L) were added to the crude product, and the mixture was stirred at 20-30°C for 12-16 hours and filtered to obtain 1-K. MS m / z: 646.4 [M+H] +

[0069] Step 10: Synthesis of compound 1-L Ethanol (705 mL) was added to a 3 L reaction vessel, followed by 1-K (352 g, 0.55 mol) and stirring. 260 mL of methylamine ethanol solution was added to the reaction vessel flask and further stirred until the reaction mixture became cloudy. The temperature of the reaction mixture was slowly raised to 35-40 °C and stirred for an additional 2-3 hours. Another 260 mL of methylamine ethanol solution was added to the reaction vessel flask and further stirred until the reaction mixture became cloudy. The reaction mixture was stirred at 35-40 °C for 20-24 hours, and the reaction mixture gradually became clear. The reaction mixture was concentrated under reduced pressure to give crude product 1-L, which was used directly in the next step without further purification. MS m / z: 645.2 [M+H] +

[0070] Step 11: Synthesis of Compound 1-M To the 1-L crude product, 704 mL of tetrahydrofuran was added. While stirring, 1100 mL of a tetrabutylammonium fluoride tetrahydrofuran solution was added to the reaction flask to clarify the reaction mixture. Then, 66 mL of anhydrous ethylenediamine was added to the reaction flask. The reaction mixture was stirred at 65-70°C for 18 hours. The reaction mixture was concentrated under reduced pressure (50°C). When the solvent could no longer evaporate, 2 L of ethyl acetate and 2 L of water were added, followed by extraction. The organic phase was washed with water (2 L x 4). The collected organic phase was concentrated under reduced pressure (50°C). When the solvent could no longer evaporate, 700 mL of water was added to a rotary evaporator to create a cloudy mixture. The mixture was transferred to a round-bottom flask and stirred at 20-30°C for 16 hours. The mixture was suction filtered under reduced pressure using a Buchner funnel, and the filter cake was rinsed with water (50 mL x 3) to obtain a solid. The solid was further stirred with acetonitrile (700 mL) at 20-30°C for 30 minutes. The mixture was filtered under reduced pressure using a Buchner funnel, and the filter cake was rinsed with acetonitrile (30 mL x 3) to obtain 1-M. MS m / z: 515.3 [M+H] +

[0071] Step 12: Synthesis of Compounds of Formula (I) Ethyl acetate (3.4 L) was added to a 10 L three-neck flask. 1-M (168 g) was added to the reaction vessel and further stirred until the reaction solution became clear. 800 mL of hydrochloric acid / ethyl acetate (4 mol / L) was slowly added to the reaction vessel flask, causing a solid to precipitate in the reaction solution. The temperature of the reaction system was gradually raised from 20-30°C to 35-40°C, and the mixture was stirred under these conditions for 19 hours. The reaction system was suction filtered under reduced pressure to obtain a yellow solid, which was rinsed with ethyl acetate (50 mL x 3). The yellow solid was added to a vessel containing 1700 mL of water with stirring, causing the system to become clear. The pH of the reaction system was adjusted to 9-10 using 2.0 L of the prepared Na2CO3 solution (1 mol / L). After a light green solid precipitated, the mixture was further stirred for 0.5-1 hour. The pale green solid was placed in a round-bottom flask containing acetonitrile (1.7 L) and stirred for 0.5 to 1 hour at 20 to 30° C. The reaction mixture was suction filtered under reduced pressure to obtain the compound of formula (I). MS m / z: 415.2 [M+H] +

[0072] Example 2: Preparation of crystalline form B of compound of formula (II) [ka]

[0073] Step 1: Synthesis of crystalline form B of compound of formula (II) EtOH (2.5 L) was added to a 5 L reaction vessel, followed by the addition of compound of formula (I) (121.2 g) and stirring. 26 mL of concentrated hydrochloric acid (12 mol / L) was slowly added to the reaction vessel flask, and the reaction solution dissolved and then became cloudy. The temperature of the reaction system was gradually raised to 40-45°C and stirred for an additional 12-16 hours. The temperature was then naturally lowered to 10-20°C, and the reaction system was subjected to suction filtration under reduced pressure using a Buchner funnel. The filter cake was washed with ethanol (50 mL x 3), collected, and vacuum dried (45°C, -0.1 MPa, 12 hours) to obtain crystalline form B of compound (II). IC-Cl analysis revealed that the hydrochloride salt (Cl) was present at 1.0 mcg. - % was confirmed to be 7.6%). MS m / z: 415.2 [M+H] +;1HNMR(400MHz,DMSO-d6)δ12.22(s,1H),10.63(s,1H),9.07(d,J=4.8Hz,1H),8.37(d,J=2.8Hz,1H),8.24(d,J=5.6Hz,1H),7.99(s,1H),7.9 3-7.90(m,2H),7.71(d,J=5.6Hz,1H)7.15(s,1H)7.00(d,J=8.8Hz,1H) ,4.23-4.19(m,1H)2.91(d,J=4.8Hz,3H)2.27(s,3H)0.79-0.67(m,4H).

[0074] Example 3: Preparation of crystalline form A of compound of formula (I) 1.6 g of crystalline form B of compound of formula (II) was weighed and dissolved in 25 mL of water. The mixture was stirred at 10-20°C. 1.88 g of sodium carbonate was dissolved in 5 mL of water, and then slowly added to the above solution and stirred for an additional 0.5 hours. The reaction mixture changed from clear to cloudy, and a solid precipitated. The mixture was filtered, and the filter cake was dried under vacuum for 12 hours to obtain crystalline form A of compound of formula (I). MS m / z: 415.2 [M+H] + .

[0075] Example 4: Preparation of crystalline form C of compound of formula (III) Crystalline Form A of compound of formula (I) (1.2 g, 2.90 mmol) was weighed and mixed with ethanol (18 mL), and concentrated HCl (12 M, 603.20 μL, 2.5 eq) was added thereto. The solid was observed to dissolve. The mixture was stirred overnight in an oil bath at 50°C, filtered, and baked to obtain crystalline Form C of compound of formula (III). IC-Cl analysis revealed that two hydrochloride salts (Cl) were present. - % was confirmed to be 18.03%).

[0076] Example 5: Study of hygroscopicity of crystalline form B of compound of formula (II) Test materials: SMS DVS Intrinsic Dynamic Vapor Sorption Apparatus Experimental Method: Samples (10 mg) are placed in the DVS sample plate for testing. Test Results: The DVS spectrum of crystalline form B of compound of formula (II) is shown in Figure 8. ΔW=0.28%. Testing Conclusion: For crystalline form B of compound of formula (II), the weight gain due to moisture absorption at 80% RH / 25° C. was 0.28%, and the sample was slightly hygroscopic.

[0077] Example 6: Testing the solid state stability of crystalline form B of compound of formula (II) Approximately 900 mg of crystalline Form B of compound of Formula (II) was weighed and placed at the bottom of a glass sample vial, which was then spread into a thin layer. The sample was completely exposed. The samples were then sampled and analyzed (XRPD) on days 10 and 30 after exposure to high temperature and humidity. The irradiated samples were placed on a clean watch glass, spread into a thin layer, and covered with a quartz glass cover. The samples were then sampled and analyzed (XRPD) on days 5 and 10. For long-term and accelerated experiments (protected from light), each sample was placed in a two-layer LDPE bag, and each layer of the LDPE bag was individually sealed. The LDPE bags were then placed in aluminum foil bags containing desiccant, which were then heat-sealed. The samples were then examined under conditions of 25°C / 60% RH, 30°C / 65% RH, and 40°C / 75% RH. The results were compared with the initial results on day 0. The test results are shown in Table 7 below. [Table 7] Conclusion: Crystalline form B of compound of formula (II) exhibits good stability under all conditions of influence factors, illumination, acceleration and long-term experiments.

[0078] Biological Test Data In this study, Promega's Luminescent Cell Viability Assay (CellTiter-Glo® method) was used to detect cell proliferation. The CellTiter-Glo® Luminescent Cell Viability Assay (CellTiter-Glo® Luminescent Cell Viability Assay Kit) is a homogeneous and rapid method for detecting the viability of live cells in culture by quantitatively measuring ATP. ATP is an indicator of viable cell metabolism. Cell lysis and the resulting luminescent signal are proportional to the amount of ATP present, which is directly proportional to the number of viable cells in culture. A higher fluorescence reading indicates a higher intracellular ATP content and therefore higher cell viability. Therefore, the degree of cell proliferation inhibition by a compound can be monitored by fluorescence values.

[0079] 1.1 Experimental design To analyze the inhibitory effect of crystalline form B of compound of formula (II) on the proliferation of BCR-FMS-BaF3 cells, three replicates were set up and the compound concentrations ranged from 10 μM to 0.0256 nM.

[0080] 1.2 Experimental steps 1.2.1 Setting the compound working solution concentration 1) Compounds were diluted to 2 mM with 100% DMSO, i.e., 5 μL of 10 mM compound stock solution was taken and placed in column 1 of a 96-well compound plate, then 20 μL of 100% DMSO was added and mixed evenly. 2) 20 μL of 100% DMSO was added to columns 2-10 of a 96-well compound plate. 3) 5 μL of compound was taken from column 1 and mixed evenly in column 2, then 5 μL of compound was taken from column 2 and mixed evenly in column 3, and the above steps were repeated up to column 9 to complete the gradient dilution of the compound. 4) Preparation of compound working solution: Transfer 2 μL of compound per well from column 2 of the compound plate to a new 96-well compound plate and add 78 μL of cell culture medium per well. Transfer 2 μL of 100% DMSO from the negative control wells and add 78 μL of cell culture medium. 5) Compound concentrations are final concentrations, blank control wells are medium wells without cells, and negative control is 0.5% DMSO.

[0081] 1.2.2 Cell inoculation and drug treatment 1) The BCR-FMS-BaF3 cell suspension was collected and centrifuged at 1000 rpm (revolutions per minute) for 5 minutes. The supernatant was removed, resuspended in preheated medium, and counted. After counting, the cell suspension was diluted with cell culture medium to the required density, as shown in Table 4-1. 80 μL of the cell suspension was inoculated into each well of a 96-well cell culture plate. 2) On the day of the experiment, 20 μL of the compound working solution was added to each well of the cell plate and incubated in a 37°C, 5% CO2 incubator for 3 days. The incubation time varies depending on the cell type. The specific cell seeding density and incubation period are shown in Table 8. [Table 8] 3) After incubation, 20 μL of CTG detection reagent was added to each well of the cell plate and incubated for 10 minutes at 25° C. After incubation, luminescence signals were detected using EnVision.

[0082] 1.3 Data analysis The data were used to calculate cell viability after compound treatment according to the following formula: % inhibition = (RFU 化合物 -RFU 陰性対照 ) / (RFU blank -RFU 陰性対照 ) x 100%. Blank reading: average reading of blank control wells, negative control: cells treated with 0.5% DMSO. Then plotted using Prism and the IC of the compound 50 values ​​were calculated.

[0083] 1.4 Experimental results: shown in Table 9. [Table 9] Experimental conclusion: Crystalline form B of compound of formula (II) can significantly inhibit the proliferation of BCR-FMS-BaF3 cells.

Claims

1. A crystal of crystalline form A of compound of formula (I) having characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 9.25±0.20°, and 14.45±0.20° in a powder X-ray diffraction pattern. 【Chemistry 1】

2. 2. The crystalline form A according to claim 1, wherein the powder X-ray diffraction pattern exhibits characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 7.98±0.20°, 9.25±0.20°, 14.45±0.20°, 16.02±0.20°, and 24.52±0.20°.

3. 3. The crystalline form A according to claim 2, wherein the powder X-ray diffraction pattern exhibits characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 7.98±0.20°, 9.25±0.20°, 12.21±0.20°, 14.45±0.20°, 16.02±0.20°, 20.20±0.20°, and 24.52±0.20°.

4. 4. The crystalline form A according to claim 3, wherein the powder X-ray diffraction pattern exhibits characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 7.98±0.20°, 9.25±0.20°, 10.08±0.20°, 12.21±0.20°, 14.45±0.20°, 15.13±0.20°, 16.02±0.20°, 20.20±0.20°, 21.35±0.20°, 21.96±0.20°, and 24.52±0.20°.

5. 5. The crystalline form A according to claim 4, wherein the powder X-ray diffraction pattern exhibits characteristic diffraction peaks at 2θ angles of 6.12°, 7.98°, 9.25°, 10.08°, 12.21°, 14.45°, 15.13°, 16.02°, 18.34°, 20.20°, 21.35°, 21.96°, 23.41°, 24.52°, 25.43°, 27.39°, 28.56°, 29.03°, 29.96°, 31.75°, and 37.38°.

6. The crystalline form A according to any one of claims 1 to 5, which has an endothermic peak at 200.9±3.0°C in a differential scanning calorimetry curve.

7. Compound of formula (II). 【Chemistry 2】

8. A crystal of crystalline form B of compound of formula (II) having characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, and 15.918±0.200° in a powder X-ray diffraction pattern. 【Transformation 3】

9. 9. The crystalline form B according to claim 8, wherein the powder X-ray diffraction pattern exhibits characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, 13.019±0.200°, 15.918±0.200°, 19.697±0.200°, and 25.200±0.200°.

10. 10. A crystalline form of crystalline form B according to claim 9, wherein the powder X-ray diffraction pattern exhibits characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, 7.941±0.200°, 11.218±0.200°, 13.019±0.200°, 15.918±0.200°, 19.697±0.200°, and 25.200±0.200°.

11. 11. A crystalline form B according to claim 10, wherein the powder X-ray diffraction pattern exhibits characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200°, 7.941±0.200°, 11.218±0.200°, 13.019±0.200°, 14.701±0.200°, 15.918±0.200°, 17.338±0.200°, 19.697±0.200°, 22.540±0.200°, 25.200±0.200°, and 27.798±0.200°.

12. The powder X-ray diffraction pattern shows the following angles: 4.579°, 6.898°, 7.941°, 9.070°, 9.423°, 11.218°, 13.019°, 13.796°, 14.701°, 15.182°, 15.918°, 17.338°, 18.939°, 19.461°, 19.697°, 20.700°, 21.602°, 22.200°, 22.540°, 22.818°, 23.919°, 24.701°, 25.182°, 26.200°, 27.200°, 28.200°, 29.200°, 30.200°, 31.200°, 32.200°, 33.200°, 34.200°, 35.200°, 36.200°, 37.200°, 38.200°, 39.200°, 40.200°, 41.200°, 42.200°, 43.200°, 44.200°, 45.200°, 46.200°, 47.200°, 48.200°, 49.200°, 50.200°, 51.200°, 52.200°, 53.200°, 54.200°, 55.200°, 56.200°, 57.200°, 58.200°, 12. The crystalline form B according to claim 11, which has characteristic diffraction peaks at 2θ angles of 4.461°, 25.200°, 25.639°, 26.563°, 27.120°, 27.798°, 28.156°, 28.595°, 29.000°, 29.339°, 30.739°, 32.064°, 32.355°, 33.201°, 34.156°, 35.020°, 36.063°, and 39.176°.

13. The crystal of crystalline form B according to any one of claims 8 to 12, wherein a differential scanning calorimetry curve shows an onset of one endothermic peak at 240.16±3.0°C and a peak value of one exothermic peak at 282.04±3.0°C.

14. The crystal of crystalline form B according to any one of claims 8 to 12, wherein the weight loss at 200.0±3°C in the thermogravimetric analysis curve reaches 0.3643%.

15. Use of a crystal of crystalline form A according to any one of claims 1 to 6, a compound according to claim 7, or a crystal of crystalline form B according to any one of claims 8 to 14 in the manufacture of a medicament for the treatment of tenosynovial giant cell tumor.

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