Crystalline form of 1H-pyrrolo[2,3-c]pyridine compound and its manufacturing method

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 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 and its crystalline form in the preparation of a medicament for treating 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 controls the growth of bone marrow progenitor cells, monocytes, and macrophages, as well as osteoclasts and dendritic cells differentiated from macrophages. To exert its biological effects, it must bind to its only 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. The binding of CSF-1 and CSF-1R at their extracellular domains induces the dimerization of CSF-1R, which in turn causes the autophosphorylation of the intracellular CSF-1R kinase domain. Once phosphorylated, CSF-1R serves as a docking site for several cytoplasmic signaling molecules, ultimately triggering a series of signaling cascade reactions. For example, phosphorylation of the tyrosine residue at position 697 of CSF-1R can activate the MAPK signaling pathway, phosphorylation of the tyrosine residue at position 721 can activate the PI3K and PLCγ signaling pathways, and so on.

[0004] Colony-stimulating factor 1 receptor (CSF-1R) is a key target in the control 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, 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 functions, 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. Thus, inhibiting macrophage survival / activation by inhibiting CSF-1 / CSF1R signaling has become an important strategy in cancer immunotherapy.

[0005] Recent studies have shown that CSF-1R inhibitors can be used in various ways in the field of disease treatment, either alone or in combination with various anti-cancer therapies such as antiangiogenesis, T cell adoptive 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 available on the market. Pexidartinib (PLX-3397), developed by Plexxikon and acquired by Daiichi Sankyo, is a dual inhibitor of CSF-1R and c-Kit, 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), characterized in that it has characteristic diffraction peaks at 2θ angles of 6.12±0.20°, 9.25±0.20° and 14.45±0.20° in an X-ray powder 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, the analytical data of 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 onset of one endothermic peak 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 a crystalline form B of compound of formula (II), characterized in that the crystalline form B has characteristic diffraction peaks at 2θ angles of 4.579±0.200°, 6.898±0.200° and 15.918±0.200° in an X-ray powder 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 23.200±0.200° .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±0.200° Characteristic diffraction peaks are present at 2θ angles of 2.064±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°, 23.02°, 24.02°, 25.01°, 26.02°, 27.02°, 28.02°, 29.02°, 30.02°, 31.02°, 32.02°, 33.02°, 34.02°, 35.02°, 36.02°, 37.02°, 38.02°, 39.02°, 40.02°, 41.02°, 42.02°, 43.02°, 44.02°, 45.02°, 46.02°, 47.02°, 48.02°, 49.02°, 50.02°, 51.02°, 52.02°, 53.02°, 54.02°, 55.02°, 56.02°, 57.02°, 58.02°, 59.02°, 60.02°, 61.02°, 62.02°, 63 Characteristic diffraction peaks exist 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 approximately as shown in FIG.

[0023] In some embodiments of the present invention, the analytical data of 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 of crystalline form B above 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 crystalline form B above has 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 a crystalline form C of compound of formula (III), characterized in that the powder X-ray diffraction pattern has characteristic diffraction peaks at 2θ angles of 5.35±0.20°, 8.66±0.20° and 14.22±0.20°. [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 approximately as shown in FIG.

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

[0035] In some embodiments of the present invention, the differential scanning calorimetry curve for crystalline form C above has an onset of one endothermic peak 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 relates to Step 1) adding a compound of formula (I) to a solvent to form a solution; 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 present invention further provides a method for producing crystalline form B of compound of formula (II), wherein the stirring time is 12 to 24 hours.

[0038] The present invention relates to Step 1) adding a compound of formula (I) to a solvent to form a solution; 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 method for producing crystalline form B of compound of formula (II) is further provided, wherein the stirring time is 12 to 16 hours.

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

[0040] [Definitions and Explanations] Unless otherwise stated, 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, but 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 ingredients.

[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 reactions of the present invention and the necessary 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 compound of the present invention can be confirmed by conventional methods known to those skilled in the art, and when the present invention involves the absolute configuration of the 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 light 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 the present invention is not limited to these examples in any way.

[0045] All of the 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; for commercially available compounds, supplier catalog names are used. Effect of the Invention

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

[0049] The 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): Name of instrument: PANalytical X'pert 3 Type X-ray Diffraction Equipment 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 Instruments 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-scattering slit: 1mm Scan range: 3~40deg Step angle: 0.02 deg Step size: 0.5 seconds Test method: Place the sample on the sample plate, flatten the surface of the sample plate, and finally put the sample plate into the X-ray diffraction device for testing.

[0052] Differential Scanning Calorimetry (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: The 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: The sample (2.745 mg) is placed in an alumina crucible and heated from 40°C to 500°C at a heating 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: Samples (10 mg) are 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 description of the drawings]

[0057] [Figure 1] FIG. 1 is an XRPD spectrum of Cu-Kα radiation of crystalline form A of compound of formula (I). [Diagram 2] 1 is a DSC spectrum of crystalline form A of compound of formula (I). [Diagram 3] FIG. 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). [Diagram 5] 1 is a TGA spectrum of crystalline form B of compound of formula (II). [Figure 6] FIG. 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 PREFERRED EMBODIMENTS

[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 thereto, and the mixture was stirred for 40 hours in an autoclave at an external temperature of 110°C under an atmosphere of carbon monoxide (2 MPa). After the reaction was completed, the mixture was filtered through diatomaceous earth and concentrated under reduced pressure to remove the solvent, and a crude product was obtained. The crude product was purified by column chromatography (n-heptane:ethyl acetate=1:1) to obtain 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), the reaction solution was cooled to 0°C, N-iodosuccinimide (351.05 g, 1.56 mol) was added thereto, and the reaction solution was stirred at 0-5°C for 1 hour. The reaction solution 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 thereto. After stirring for 1 hour, the product 1-C was obtained by suction filtration. MS m / z: 302.7 [M+H] +

[0062] Step 3: Synthesis of compound 1-D 1-C (200g, 662.10mmol) was dissolved in tetrahydrofuran (3L), the reaction system was cooled to 0-5℃, sodium hydride (39.73g, 993.15mmol, 60% content) was slowly added thereto, and after the addition was completed, stirring was continued at 0-5℃ for 0.5 hours, 2-(chloromethoxy)ethyltrimethylsilane (139.09g, 834.25mmol) was added thereto, the reaction system was naturally heated to 20℃, and stirring was continued for 1 hour. Under a weak nitrogen flow atmosphere, the reaction liquid was slowly poured into 3L of water, stirred and quenched. Ethyl acetate (3L) was added for extraction, the organic phase was washed with water (3L), and further washed with saturated saline (3L), the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to remove the solvent, and a crude product was obtained. 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 the mixture was 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, cyclopropanol (1.14 kg, 19.71 mol) was added dropwise, and after the completion of the addition, the reaction solution was slowly heated to 25-30°C and stirred for 16 hours. The reaction solution was filtered under reduced pressure, and the obtained filter cake was rinsed twice with tetrahydrofuran (1 L), and the obtained filtrate was combined. The obtained filtrate was slowly poured into water (36 L), stirred for 30 minutes, and then filtered under reduced pressure to obtain a filter cake. Water (5 L) was added to the obtained filter cake, stirred for 30 minutes, and then 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), to which Pd / C (5 g, 555.06 umol, 10% content) was added, and the reaction was stirred under hydrogen atmosphere (30-40 psi) at 25-30° C. for 16 h. The reaction was filtered through diatomaceous earth, avoiding vacuum pumping to dryness, and the filtrate was rotary evaporated under reduced pressure to remove the solvent to give 1-G. MS m / z: 151.2 [M+H] +

[0065] Step 6: Synthesis of compound 1-H 1-G (154g, 1.03mol) was dissolved in 1,4-dioxane (2.25L) and water (45mL), and 2,5-dibromo-4-methylpyridine (308.77g, 1.23mol), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (29.67g, 51.27mmol), potassium carbonate (354.31g, 2.56mol) were added thereto, and the system was replaced with nitrogen three times. Tris(dibenzylideneacetone)dipalladium(0) (14.74g, 25.64mmol) was added thereto, and the system was replaced with nitrogen three times. The reaction solution was stirred at 55-60°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: about 6 L) was concentrated under reduced pressure to a volume of about 3.5 L, slowly poured into n-heptane (17.5 L), stirred for 0.5 hours, and then filtered under reduced pressure with suction to obtain a filter cake. Water (3 L) was added thereto, and the mixture was stirred for 20 minutes. The mixture was then filtered under reduced pressure with suction 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), 4-dimethylaminopyridine (41.56 g, 340.21 mmol) was added thereto, and di-tert-butyl dicarbonate (1.93 kg, 8.85 mol) was added dropwise, and the reaction solution was stirred at an external temperature of 30 to 40 ° C for 3 hours. The reaction solution was washed with water (10 L), and the organic phase was washed with saturated saline (10 L). The organic phase was collected and dried over anhydrous sodium sulfate (1 kg), and then concentrated under reduced pressure to obtain a crude product. n-Heptane (5 L) was added to the crude product, and the mixture was stirred at 0 ° C for 15 minutes in an ice bath. When a solid gradually precipitated, the mixture was stirred for another hour, and then suction filtered to obtain 1-I. MS m / z: 420.1 [M+H] +

[0067] Step 8: Synthesis of compound 1-J 1-I (500.31g, 1.19mol), bis(pinacolato)diboron (392.81g, 1.55mol), dioxane (5L), potassium acetate (236.73g, 2.38mol) were added to the reaction vessel in this order, and the reaction system was replaced with nitrogen three times. [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (43.71g, 0.08mol) was added thereto, and the reaction system was replaced with nitrogen three times. After that, the reaction system was protected with a nitrogen flow, and the reaction system was heated to 70-80℃ 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), and the filtrates were combined and concentrated. 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 (385.26g, 0.89mol), 1-J (466.95g, 0.91mol), dioxane (3.85L) were added to the reaction vessel in order and stirred, potassium phosphate (385.81g, 1.82mol) was dissolved in water (385mL), and then added to the reaction vessel at room temperature (20-30℃), the reaction system was replaced with nitrogen three times, the reaction system was cooled to 0-5℃, [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride dichloromethane complex (65.25g, 0.09mol) was added thereto, and the reaction system was replaced with nitrogen three times. The reaction system was heated to 35-40℃ and stirred for 12-16 hours. Ethyl acetate (8L) and water (8L) were added to the reaction liquid, and the mixture was stirred for 10 minutes, then left to stand for liquid separation, and the aqueous phase was separated. The organic phase was washed with water (8 L), washed with 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, to which ethyl acetate (7.7 L) and powdered activated carbon (193.38 g) were added, and the mixture was refluxed and stirred (75-80°C) for 1 hour, then cooled to 55-60°C, and suction filtered under reduced pressure through diatomaceous earth (500 g). The filter cake was rinsed with ethyl acetate (15 L), and the filtrate was combined and concentrated under reduced pressure to about 2 L (temperature was 45°C or less), to which ethyl acetate (0.3 L) and n-heptane (3 L) were added, 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 the 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 became cloudy. The temperature of the reaction was slowly raised to 35-40°C and stirred for another 2-3 hours. Another 260 mL of methylamine ethanol solution was added to the reaction vessel flask and further stirred until the reaction became cloudy. The reaction was stirred at 35-40°C for 20-24 hours, and the reaction gradually became clear. The reaction was concentrated under reduced pressure to give crude product 1-L, which was used directly in the next step reaction without further purification. MS m / z: 645.2 [M+H] +

[0070] Step 11: Synthesis of compound 1-M 704 mL of tetrahydrofuran was added to the 1-L crude product, and 1100 mL of tetrabutylammonium fluoride tetrahydrofuran solution was added to the reaction flask while stirring to make the reaction liquid clear. Then, 66 mL of anhydrous ethylenediamine was added to the reaction flask. The reaction liquid was stirred at 65-70°C for 18 hours. The reaction liquid was concentrated under reduced pressure (50°C), and when the solvent could not be evaporated, 2 L of ethyl acetate and 2 L of water were added, and the organic phase was washed with water (2 L x 4). The organic phase was collected and concentrated under reduced pressure (50°C), and when the solvent could not be evaporated, 700 mL of water was added to a rotary evaporator to make it cloudy, and the mixture was transferred to a round-bottom flask and stirred for 16 hours at 20-30°C. The mixture was filtered by suction 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 Compound 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 to make the reaction solution clear. 800 mL of hydrochloric acid / ethyl acetate (4 mol / L) was slowly added to the reaction vessel flask, and a solid precipitated in the reaction solution. The temperature of the reaction system was gradually raised from 20-30°C to 35-40°C, and stirred under these conditions for 19 hours. The reaction solution was suction filtered under reduced pressure to obtain a yellow solid, and the solid was rinsed with ethyl acetate (50 mL x 3). The yellow solid was added to a vessel containing 1700 mL of water while stirring, and the system became clear. The above reaction system was adjusted to pH = 9-10 using 2.0 L of the prepared Na2CO3 solution (1 mol / L), and when a light green solid precipitated, it was further stirred for 0.5-1 hour. The light 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 filtered under reduced pressure with suction 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, then the compound of formula (I) (121.2 g) was added and stirred, and 26 mL of concentrated hydrochloric acid (12 mol / L) was slowly added to the reaction vessel flask, whereupon the reaction solution dissolved and then became cloudy. The temperature of the reaction system was gradually raised to 40-45°C, and the reaction solution was stirred for another 12-16 hours. The temperature was then naturally lowered to 10-20°C, and the reaction solution was filtered by suction under reduced pressure using a Buchner funnel, the filter cake was washed with ethanol (50 mL x 3), the filter cake was collected, and the obtained filter cake was vacuum dried (45°C, -0.1 MPa, 12 hours) to obtain crystalline form B of compound (II). The IC-Cl measurement revealed that 1.0 unit of hydrochloride (Cl - % 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 the crystalline form B of the compound of formula (II) was weighed out, dissolved in 25 mL of water, and 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 another 0.5 hours. The reaction solution became cloudy instead of clear, and a solid precipitated. After filtration, the filter cake was vacuum dried for 12 hours to obtain the crystalline form A of the compound of formula (I). MS m / z: 415.2 [M+H] + .

[0075] Example 4: Preparation of crystalline form C of compound of formula (III) 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 Form C of compound of formula (III). IC-Cl measurement showed that two hydrochloride salts (Cl - % was confirmed to be 18.03%).

[0076] Example 5: Study of the hygroscopicity of crystalline form B of compound of formula (II) Test materials: SMS DVS Intrinsic Dynamic Vapor Sorption Apparatus Testing 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%. Test 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, placed on the bottom of a glass sample bottle, and spread into a thin layer. The sample was completely exposed. The samples placed in high temperature and high humidity were sampled and detected (XRPD) on the 10th and 30th days. The irradiated samples were placed on a clean watch glass, spread into a thin layer, covered with a quartz glass cover, and sampled and detected (XRPD) on the 5th and 10th days. For long-term experiments and accelerated tests (protected from light), each sample was placed in a two-layer LDPE bag, and each layer of the LDPE bag was individually fastened and sealed. The LDPE bags were then placed in aluminum foil bags containing desiccant, heat-sealed, and examined under conditions of 25°C / 60% RH, 30°C / 65% RH, and 40°C / 75% RH, and the detection results were compared with the initial detection results on day 0, and the test results are shown in Table 7 below. [Table 7] Conclusion: The crystalline form B of compound of formula (II) exhibits good stability under all conditions of influence factors, illumination, accelerated 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 is a homogeneous and rapid method to detect the viability of live cells in culture by quantitatively measuring ATP. ATP is one of the indicators of the metabolism of live cells. Cell lysis and the resulting luminescent signal are proportional to the amount of ATP present, which is directly proportional to the number of live cells in the culture. A higher fluorescence reading indicates a higher ATP content in the cells and also indicates a higher cell viability. Therefore, the degree of inhibition of cell proliferation by a compound can be monitored by the fluorescence value.

[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 concentration range of the compound was 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., take 5 μL of 10 mM compound stock solution and place into column 1 of a 96-well compound plate, then add 20 μL of 100% DMSO and mix evenly. 2) Add 20 μL of 100% DMSO to columns 2-10 of a 96-well compound plate. 3) Take 5 μL of compound from column 1 and mix evenly in column 2, then take 5 μL of compound from column 2 and mix evenly in column 3, and repeat the above steps up to column 9 to complete the gradient dilution of compound. 4) Preparation of compound working solution: Take 2 μL of compound per well from column 2 of the compound plate into a new 96-well compound plate and add 78 μL of cell media per well. Take 2 μL of 100% DMSO from the negative control wells and add 78 μL of cell media. 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) BCR-FMS-BaF3 cell suspension was collected and centrifuged at 1000 rpm (revolutions per minute) for 5 minutes, the supernatant was removed, and the cells were resuspended in pre-warmed medium and counted. After counting, the cell suspension was diluted with cell medium to the required density, and each cell density is 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 for 3 days in a 37°C, 5% CO2 incubator. Different cells require different incubation times. The specific cell seeding density and incubation days 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, the luminescence signal was detected using EnVision.

[0082] 1.3 Data analysis The data was 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 ) × 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 compounds 50 values ​​were calculated.

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

Claims

1. Crystalline form A of compound of formula (I) has 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. The crystalline form A according to claim 1, wherein the powder X-ray diffraction pattern 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°.

3. The crystalline form A according to claim 2, wherein the powder X-ray diffraction pattern 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°.

4. The crystalline form A according to claim 3, wherein the powder X-ray diffraction pattern 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°.

5. 5. The crystalline form A according to claim 4, wherein the powder X-ray diffraction pattern 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°.

6. 6. The crystalline form A of claim 5, having an XRPD pattern substantially as shown in FIG.

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

8. 8. The crystalline form A of claim 7, having a DSC pattern as shown in FIG.

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

10. Crystal form B of compound of formula (II) has 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. 【Chemistry 3】

11. The crystalline form B according to claim 10, wherein the powder X-ray diffraction pattern 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°.

12. 12. The crystalline form B of claim 11, wherein the powder X-ray diffraction pattern 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°.

13. 13. The crystalline form B of claim 12, wherein the powder X-ray diffraction pattern 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°.

14. Powder X-ray diffraction pattern: 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° 14. The crystalline form B of claim 13, having characteristic diffraction peaks at 2θ angles of 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°.

15. 15. The crystalline form B of claim 14, having an XRPD pattern substantially as shown in Figure 3.

16. The crystalline form B according to any one of claims 10 to 15, wherein a differential scanning calorimetry curve has 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.

17. 17. The crystalline form B of claim 16, whose DSC pattern is shown in Figure 4.

18. The crystalline form B according to any one of claims 10 to 15, wherein the weight loss at 200.0±3°C in the thermogravimetric analysis curve reaches 0.3643%.

19. 19. The crystalline form B of claim 18, having a TGA pattern as shown in Figure 5.

20. Use of crystalline form A according to any one of claims 1 to 8, a compound according to claim 9, or crystalline form B according to any one of claims 10 to 19 in the manufacture of a medicament relating to the treatment of tenosynovial giant cell tumor.

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