Crystalline form of fluorine-substituted pyridopyrazole compound and its preparation method

Abstract

The present invention further includes a crystalline form of a fluorine-substituted pyridopyrazole compound and a method for preparing the same, and further includes the use of the compound and the crystalline form thereof in the manufacture of a medicament for treating related diseases. [Formula 1] TIFF2024511614000011.tif63154

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to CN202110309088.5, filed on March 23, 2021. [Technical field]

[0002] The present invention relates to crystalline forms of fluorine-substituted pyridopyrazole compounds and their preparation methods, as well as the use of said compounds and their crystalline forms in the manufacture of medicaments for treating related diseases. [Background technology]

[0003] BTK is a key kinase in the B cell antigen receptor (BCR) signaling pathway. BTK irreversible inhibitors covalently bind to the kinase's active site, Cys-481, and inhibit BTK activity, thereby effectively suppressing B cell hyperproliferation and achieving anti-tumor and anti-inflammatory effects. Summary of the Invention [Problem to be solved by the invention]

[0004] Among the drugs already on the market, ibrutinib is an irreversible BTK inhibitor jointly developed by Pharmacyclis and Johnson & Johnson, and has been approved by the FDA for use in the treatment of mantle cell lymphoma, chronic lymphocytic leukemia, Waldenstrom's macroglobulinemia, and chronic graft-versus-host disease. However, ibrutinib has a relatively strong inhibitory effect on other kinases besides BTK, and inhibition of kinases such as EGFR, ITK, and TEC in particular can cause adverse reactions such as severe skin rash, diarrhea, and bleeding. Therefore, there is a need in this field to develop novel BTK inhibitors that are highly active while having good selectivity for the treatment of related diseases. [Means for solving the problem]

[0005] The present invention provides crystalline form A of compound of formula (I), characterized in that it has characteristic diffraction peaks at 2θ angles of 17.7805±0.2000°, 22.0193±0.2000°, and 27.4192±0.2000° in a powder X-ray diffraction pattern. [ka]

[0006] In some embodiments of the present invention, the crystalline form A has characteristic diffraction peaks at 2θ angles of 15.0010±0.2000°, 17.0603±0.2000°, 17.7805±0.2000°, 22.0193±0.2000°, 23.5013±0.2000°, and 27.4192±0.2000° in a powder X-ray diffraction pattern.

[0007] In some embodiments of the present invention, the crystalline form A has characteristic diffraction peaks at 2θ angles of 8.1387±0.2000°, 15.0010±0.2000°, 16.1591±0.2000°, 17.0603±0.2000°, 17.7805±0.2000°, 22.0193±0.2000°, 23.5013±0.2000°, and 27.4192±0.2000° 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 8.1387±0.2000°, 13.6809±0.2000°, 15.0010±0.2000°, 16.1591±0.2000°, 17.0603±0.2000°, 17.7805±0.2000°, 22.0193±0.2000°, 23.5013±0.2000°, 24.0218±0.2000°, and 27.4192±0.2000° in a powder X-ray diffraction pattern.

[0009] In some embodiments of the present invention, the crystalline form A has an X-ray powder diffraction pattern of 17.7805±0.2000°, 22.0193±0.2000°, and / or 27.4192±0.2000°, and / or 8.1387±0.2000°, and / or 9.8194±0.2000°, and / or 13.6809±0.2000°, and / or 1 5.0010±0.2000°, and / or 16.1591±0.2000°, and / or 17.0603±0.2000°, and / or 18.4386±0.2000°, and / or 19.5400±0.2000°, and / or 21.3193±0.2000°, and / or 23.1597±0.2000°, and / or 23.5013±0.200 0°, and / or 24.0218±0.2000°, and / or 26.1791±0.2000°, and / or 26.6006±0.2000°, and / or 27.1199±0.2000°, and / or 29.1595±0.2000°, and / or 29.7190±0.2000°, and / or 30.8417±0.2000°, and / or 31. and / or 32.9612±0.2000°, and / or 33.7773±0.2000°, and / or 34.3779±0.2000°, and / or 35.1796±0.2000°, and / or 37.1408±0.2000°.

[0010] In some embodiments of the present invention, the crystalline form A has the following X-ray powder diffraction patterns: 8.1387°, 9.8194°, 13.6809°, 15.0010°, 16.1591°, 17.0603°, 17.7805°, 18.4386°, 19.5400°, 21.3193°, 22.0193°, 23.1597°, 23.5013°, 24.0193°, 25.0193°, 26.0193°, 27.0193°, 28.0193°, 29.0193°, 30.0193°, 31.0193°, 32.0193°, 33.0193°, 34.0193°, 35.0193°, 36.0193°, 37.0193°, 38.0193°, 39.0193°, 40.0193°, 41.0193°, 42.0193°, 43.0193°, 44.0193°, 45.0193°, 46.0193°, 47.0193°, 48.0193°, 49.0193°, 50.0193°, 51.0193°, 52.0193°, 53.0193°, 54.0193°, 55.0193°, 56.0193°, 57.0193°, 58.0193°, 59.0193°, It has characteristic diffraction peaks at 2θ angles of 4.0218°, 26.1791°, 26.6006°, 27.1199°, 27.4192°, 29.1595°, 29.7190°, 30.8417°, 31.2196°, 32.2992°, 32.9612°, 33.7773°, 34.3779°, 35.1796°, and 37.1408°.

[0011] In some embodiments of the present invention, crystalline form A above has an XRPD pattern essentially as shown in FIG.

[0012] In some embodiments of the present invention, the crystalline form A has XRPD pattern analysis data as shown in Table 1. [Table 1]

[0013] In some embodiments of the present invention, the crystalline form A has an endothermic peak at 107.89±3.00° C. and an exothermic peak at 147.76±3.00° C. in a differential scanning calorimeter curve.

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

[0015] In some embodiments of the present invention, the crystalline form A exhibits a weight loss of 0.06% at 200.0±3° C. in a thermogravimetric analysis curve.

[0016] In some embodiments of the present invention, the crystalline form A is shown above, the TGA pattern of which is shown in FIG.

[0017] In some embodiments of the invention, the crystalline form A has single crystal X-ray diffraction data for a monoclinic system in space group C2 with lattice parameters a=22.0128(6) Å, b=12.7542(3) Å, c=16.0152(4) Å, α=γ=90°, β=90.4900(10)°, and volume V=4496.2(2) Å. 3 and the absolute configuration parameter Flack value is 0.04(3).

[0018] The present invention also provides Step 1) adding a compound of formula (I) to a solvent to form a solution; 2) adding an anti-solvent to the solution, stirring at a predetermined temperature for a predetermined time, filtering, and drying the filter cake under reduced pressure; the solvent is an ester-based solvent, an ether-based solvent, or an alcohol-based solvent, the anti-solvent is n-heptane, n-hexane or water; The stirring temperature is 0 to 40° C. The stirring time is 12 to 48 hours.

[0019] The present invention also provides Step 1) adding a compound of formula (I) to a solvent to form a solution; 2) adding an anti-solvent to the solution, stirring at 30° C., filtering, and drying the filter cake under reduced pressure; the solvent is ethyl acetate, tetrahydrofuran or ethanol; The antisolvent is n-heptane, n-hexane or water; The stirring time is 12 to 20 hours.

[0020] The present invention also provides the use of the above crystalline form A in the manufacture of a medicament for treating a BTK-associated disease.

[0021] In some forms of the invention, the BTK-associated disease is a hematologic tumor or an autoimmune disease.

[0022] Definitions and Explanations Unless otherwise specified, the following terms and phrases as used herein are intended to include the following meanings: Certain words and terms should not be regarded as uncertain or indefinite without specific definition, but should be understood in their general sense. When trade names are used herein, they are intended to refer to the corresponding commercial product or its active ingredients.

[0023] The intermediate compounds of the present invention may be prepared by a variety of synthetic methods familiar to those skilled in the art, including the specific embodiments listed below, embodiments combining those embodiments with other chemical synthetic methods, and equivalent alternatives familiar to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present invention.

[0024] The chemical reactions of the specific embodiments of the present invention are carried out in suitable solvents suitable for the chemical transformations of the present invention and the reagents and materials required therefor. In order to obtain the compounds of the present invention, those skilled in the art may need to modify or select synthetic steps or reaction flows based on existing embodiments.

[0025] The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art, and when the present invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed by conventional means of those skilled in the art. For example, in single crystal X-ray diffraction (SXRD), the diffraction intensity data of the cultivated single crystal is collected by a Bruker D8venture diffractometer with a CuKα radiation source and a φ / ω scanning method, and the relevant data is collected, and then the crystal structure is analyzed by a direct method (Shelxs97) to confirm the absolute configuration.

[0026] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

[0027] All solvents used in this invention are commercially available and can be used without further purification.

[0028] The solvents used in the present invention are commercially available.

[0029] Compounds are named according to conventional naming principles in the art or using ChemDraw® software; commercially available compounds are named from the supplier's catalog. Effect of the Invention

[0030] The compound of the present invention has a stable crystal form, is slightly hygroscopic, and is not easily affected by light or heat. The compound of the present invention is a BTK kinase inhibitor with high activity and excellent selectivity, and has excellent selectivity against EGFR, ITK, and TEC kinases.

[0031] The X-ray powder diffractometer (XRPD) method of the present invention Equipment name: X-ray diffraction equipment Equipment model: DX-2700BH Equipment marker: Dandong Haoyuan Ceremony Co., Ltd. Method parameters: Light tube: Cu, k-Alphal (λ=1.54059Å) Light tube voltage: 40kV, Light tube current: 40mA Divergence slit: 0.3 mm Detector slit: 1mm Anti-scattering slit: 1mm Scanning range: 3~40deg Step diameter: 0.02 deg Step length: 0.5 seconds Test 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.

[0032] Differential Scanning Calorimeter (DSC) Method of the Present Invention Instrument model: DSC1 Differential Scanning Calorimeter Test method: The sample (2.97 mg) is placed in a DSC high-pressure crucible, pressed and sealed, and then the test is performed by heating the sample from 40°C to 350°C at a heating rate of 10°C / min.

[0033] Thermogravimetric Analysis (Thermal Gravimetric Analyzer, TGA) Method of the Present Invention Instrument model: Mettler TGA2SF / 1100 Test method: The test is performed by placing a sample (2-5 mg) in a TGA alumina crucible, heating the sample from 40°C to 500°C at a heating rate of 10°C / min under conditions of 20 mL / min N2, and the weight loss from 40°C to 200°C is 0.06%.

[0034] Dynamic Vapor Sorption (DVS) Method of the Present Invention Instrument Model: SMSDVS IntrinsicPlus Dynamic Vapor Sorption Apparatus Test conditions: Place the sample (10-20 mg) in the DVS sample tray and perform the test. The DVS parameters are explained in more detail below. Temperature: 25℃ Equilibrium: dm / dt=0.002% / min (minimum: 10min, maximum: 180min) Drying: Dry at 0% RH until dm / dt≦0.002% / min or for a maximum of 180 min. RH(%) test gradient: 10%(0%~90%), 5%(90~95%) RH(%) test gradient range: 0%-95%-0%

[0035] [Table 2] [Brief description of the drawings]

[0036] [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] 1 is a TGA spectrum of crystalline form A of compound of formula (I). [Figure 4] 1 is a DVS spectrum of crystalline form A of compound of formula (I). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0037] In order to better understand the present invention, the present invention will be further described below with reference to specific examples, which are not intended to limit the present invention.

[0038] Example 1: Preparation of compound of formula (I) [ka]

[0039] Step 1: Synthesis of compound b Preparation of sodium hypochlorite solution: NaHCO3 (11.00g, 130.94mmol, 7.05e-2eq) was dissolved in NaClO (2.42kg, 2.60mol, 2L, 8% purity, 1.40eq). KBr (2M, 96.00mL, 1.03e-1eq) (aqueous solution) was added to a dichloromethane (1.6L) solution of compound a (400g, 1.86mol, 1eq) and 2,2,6,6-tetramethylpiperidine oxide (2.98g, 18.97mmol, 1.02e-2eq), and the mixture was kept at 5-35°C in an ice bath. The sodium hypochlorite solution was added dropwise, and after the dropwise addition, the mixture was kept at 25-35°C and the reaction was continued for 30min. After the reaction mixture was allowed to stand, the liquids were separated, and the separated aqueous phase was extracted with dichloromethane (1.6 L). The combined organic phase was washed with 1M hydrochloric acid (containing 1.6 L of KI (616.85 g, 3.72 mol, 2 eq)) and 10% sodium thiosulfate solution (1.6 L) in that order, and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to dryness to obtain compound b. LCMS (ESI): m / z (M-55) + : 158.7.

[0040] Step 2: Synthesis of compound d In a dry ice ethanol bath and under nitrogen protection, n-butyllithium (2.5M, 1.13L, 1.07eq) was added dropwise to a 2.1L solution of compound c (309.65g, 2.69mol, 1.02eq) in tetrahydrofuran while keeping at -68 to -40°C. After completion of the dropwise addition, the mixture was reacted at -68 to -40°C for 1 hour. A solution of compound b (563g, 2.64mol, 1eq) in tetrahydrofuran (0.7L) was added dropwise while keeping at -68 to -40°C. After completion of the dropwise addition, the mixture was slowly heated to 0°C and reacted for 2 hours. Saturated ammonium chloride solution (1.2L) was added to the reaction solution while stirring, and the reaction solution was concentrated under reduced pressure to about 3L, extracted with ethyl acetate (1.2L*3), and the combined organic phase was washed with 1M hydrochloric acid (1.2L) and half-saturated brine (1.2L) in that order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness. Compound d was obtained by concentration. LCMS (ESI): m / z (M-55) + : 272.9.

[0041] Step 3: Synthesis of compound e Compound d (464g, 1.41mol, 1eq) and 2,2,6,6-tetramethylpiperidine oxide (2.23g, 14.16mmol, 0.01eq) in dichloromethane (1.8L) were added with KBr (2M, 74.00mL, 1.05e-1eq) (aqueous solution), and the mixture was kept at 10-15°C in an ice bath. NaClO (1.84kg, 1.98mol, 1.72L, 8% purity, 1.4eq) was added dropwise, and the mixture was allowed to react for 10min while keeping the temperature at 10-15°C. NaHCO3 (40g, 476.15mmol, 3.37e-1eq) was replenished, and the reaction was continued for 30min. The reaction solution was left to stand, separated, and the separated aqueous phase was extracted with dichloromethane (9 L). The combined organic phase was washed with 1M hydrochloric acid (containing 2.35 kg KI, 2 eq, 9 L) and 10% sodium thiosulfate solution (9 L) in that order, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness. Analysis by supercritical fluid chromatography detection (column: Chiralpak AD-3150 x 4.6 mm I.D, 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B is 5% to 40% within 5.5 min, held at 5% for 1.2 min; flow rate: 2.5 mL / min; column temperature: 40 °C; wavelength: 220 nm) revealed that it was a single-configuration compound. Compound e was obtained. LCMS (ESI): m / z (M-55) + : 271.1.

[0042] [ka]

[0043] Step 4: Synthesis of compound h While stirring, K2CO3 (1.47 kg, 10.63 mol, 1.5 eq) was added to a solution of compound g (1 kg, 7.09 mol, 751.88 mL, 1 eq) and compound f (1.11 kg, 8.50 mol, 1.2 eq) in acetonitrile (5 L), and the temperature was raised to 75 ° C. and reacted for 13 hr. The reaction solution was cooled to room temperature, and after standing, the supernatant was added to 50 L of water, and after 10 min, a large amount of solid was generated, and the mixture was stirred for another 10 min. The remaining reaction solution was filtered, the filter cake was washed with acetonitrile (1 L), and the combined filtrate was added to 10 L of water while stirring, and after 10 min, a large amount of solid was generated, and the mixture was stirred for another 10 min. The suspensions of the above two batches were combined and filtered, the filter cake was washed with water (1 L), and the filter cake was collected by suction filtration until dryness. Compound h was obtained. 1 H NMR (400MHz, CDCl3) δ 8.20 - 8.23 ​​(m, 2H),7.11 - 7.14 (m, 2H),7.02-7.04 (m, 2H),6.96-7.01 (m, 1H).

[0044] Step 5: Synthesis of compound i In a hydrogen water generating bottle, Pd / C (4 g, 10% purity) (wet palladium carbon) was added to a solution of compound h (200 g, 796.22 mmol, 1 eq) in methanol (1 L), and the mixture was purged with argon three times and hydrogen gas three times. The mixture was reacted at hydrogen gas pressure (30 psi) and room temperature (25° C.) for 16 hours. The reaction solution was filtered (through diatomaceous earth), the filter cake was washed with methanol (1 L), and the combined filtrate was concentrated under reduced pressure to dryness to obtain compound i. LCMS (ESI): m / z(M+1) + : 222.0.

[0045] Step 6: Synthesis of compound j Compound i (350 g, 1.58 mol, 1 eq) was added to concentrated hydrochloric acid (2 L) and water (1 L) in an ice-water bath (5°C). To the resulting suspension, a solution of NaNO2 (218.35 g, 3.16 mol, 2 eq) in water (0.5 L) was added dropwise for 0.5 hr while keeping the temperature at 5-15°C. After the addition was completed, the mixture was kept at 5-10°C and reacted for 1 hr. A solution of SnCl2·2H2O (1.43 kg, 6.33 mol, 4.00 eq) in concentrated hydrochloric acid (1 L) was added dropwise to the reaction mixture while keeping the internal temperature at 5-15°C for a total of about 3 hr. After the addition was completed, water (0.75 L) was added to distribute the solid uniformly in the reaction system. The mixture was slowly heated to room temperature (25°C) and the reaction was continued for 4 hr. The reaction solution was filtered, the filter cake was washed with water (3L), the filter cake was collected, the filter cake was divided into 4 fractions, each fraction was dispersed in a mixed solution of methanol (0.5L) and dichloromethane (4L), the pH was adjusted to 14 using 6M NaOH solution, and the mixture was allowed to stand and separated. The aqueous phase was extracted with dichloromethane (2L*2), and the organic phase was collected. The organic phases were combined and filtered, the filter cake was washed with 500mL of dichloromethane, the combined filtrate was separated, the aqueous phase was extracted with dichloromethane (1L*2), and the organic phase was collected. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness. Compound j was obtained. LCMS (ESI): m / z(M+1) + : 237.1.

[0046] Step 7: Synthesis of compound k AcOH (15.21 mol, 870.00 mL, 4.96 eq) was added to a solution of compound e (1 kg, 3.06 mol, 1 eq) and compound j (1 kg, 4.23 mol, 1.38 eq) in ethanol (5 L), and the mixture was reacted at room temperature (25°C) for 12 hours. The reaction solution was concentrated under reduced pressure to dryness. Compound k was obtained.

[0047] Step 8: Synthesis of compound l Cs2CO3 (2.8 kg, 8.59 mol, 2.46 eq) was added to a solution of compound k (1.9 kg, 3.49 mol, 1 eq) in N,N-dimethylformamide (6.5 L), and the mixture was heated to 100°C and reacted for 1.5 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to dryness using an oil pump. The filter cake was washed with dichloromethane (1 L), and the combined filtrate was successively concentrated under reduced pressure to dryness using a water pump and an oil pump. The concentrate was dispersed in ethanol (8 L), stirred for 1 hour, filtered, the filter cake was washed with 1 L of ethanol, and the combined filtrate was concentrated under reduced pressure to dryness, purified by a silica gel column (petroleum ether: ethyl acetate = 1:0 to 4:1, 10% dichloromethane added), and the obtained concentrate was dispersed in ethyl acetate (1.1 L), and 3.3 L of petroleum ether was added while stirring, and the mixture was stirred at room temperature (25°C) for 3 hours. Filtration was performed, and the filter cake was washed with 200 mL of petroleum ether. The filter cake was collected, concentrated under reduced pressure, and dried. Compound 1 was obtained. LCMS (ESI): m / z: 525.3 [M+1]. Supercritical fluid chromatography detection (column: Chiralpak AD-350 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% in 5 min, 0.5% to 10% in 0.5 min) was used. 40% Analysis by a 2.5 mL / min column (from 0.01 to 5%, 5% held for 1.5 min; flow rate: 2.5 mL / min; column temperature: 35° C.; wavelength: 220 nm) showed that the compound was racemic.

[0048] Step 9: Synthesis of compound m Compound l (1.26 kg, 2.40 mol, 1 eq) was dissolved in methanol (2.6 L) and HCl (4 M, 3.80 L, 6.33 eq) was added dropwise to the suspension in methanol (2.6 L). After the addition, the mixture was reacted at room temperature (25 ° C) for 0.5 hr, at which point the solution was clear. The reaction solution was concentrated under reduced pressure until dry. The concentrate was dissolved in a mixed solution of 4 L of dichloromethane and 400 mL of methanol, and the pH of the solution was adjusted to 14 using 6 M sodium hydroxide solution. The mixture was stirred for another 0.5 hr, allowed to stand and separated, and the aqueous phase was extracted with 3 L of dichloromethane. The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure until dry. Supercritical fluid chromatography detection (column: Chiralpak IG-350 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B 5% to 40% in 2 min, 40% Analysis by 1.2 min at 5% and 0.8 min at 5%; flow rate: 4mL / min; column temperature: 35℃; wavelength: 220nm) showed it to be racemic, which was purified by SFC separation (column: DAICELCHIRALPAKAD (250mm*50mm,10μm); mobile phase: A: supercritical carbon dioxide, B: [0.1%NH3H2OMEOH]; B%: 50%~50%,min) to give m (retention time 2.584min). LCMS(ESI): m / z: 425.2[M+1].

[0049] Step 10: Synthesis of Compound of Formula (I) To a solution of compound m (280 g, 659.73 mmol, 1 eq) in tetrahydrofuran (2.8 L) and water (2.8 L), Na2CO3 (105 g, 990.66 mmol, 1.5 eq) was added, and a solution of acryloyl chloride (65.88 g, 727.92 mmol, 59.35 mL, 1.1 eq) in tetrahydrofuran (0.7 L) was added dropwise to the reaction solution, and the reaction was carried out at room temperature (28 °C) for 0.5 hours. After adding a solution of acryloyl chloride (10.6g, 117.12mmol, 9.55mL, 1.78e-1eq) in tetrahydrofuran (50mL) and Na2CO3 (70g, 660.45mmol, 1.00eq), the reaction was continued at room temperature (28℃) for 0.5hr, the reaction solution was adjusted to pH 6 with 1N hydrochloric acid, extracted with dichloromethane (3.5L*2), the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness. The concentrate was purified by silica gel column (petroleum ether:ethyl acetate=1:0-1:1, 10% dichloromethane was added to promote dissolution.). The crude product obtained was dissolved in dichloromethane (1L), concentrated under reduced pressure until dryness, and this operation was repeated three times. The mixture was then transferred to an oil pump and concentrated under reduced pressure to obtain the compound of formula (I). The compound was analyzed by supercritical fluid chromatography detection (column: Cellulose2150×4.6mmI.D, 5μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B is from 5% to 40% within 5 minutes, held at 40% for 2.5 minutes, returned to 5% and equilibrated for 2.5 minutes; flow rate: 2.5mL / min; column temperature: 35℃; wavelength: 220nm), and a single configuration compound (retention time 6.916min) was obtained. 1HNMR(400MHz,DMSO-d6) δ 9.08(s,1H),8.32(s,1H),7.85(d,J=8.8Hz,2H),7.24-7.33(m,4H),7.09-7.11(m,1H),6.8 0-6.7582,1H),6.08(t,J=16.1Hz,1H),5.59-5.70(m,1H),4.70-4.72(m,0.5H),4.32(t,J= 12.7Hz,1H),4.07-4.10(m,0.5H),3.57-3.44(m,0.5H),3.33-3.15(m,1.5H),3.12-3.03(m ,0.5H),3.03-2.88(m,0.5H),2.27-2.16(m,1H),2.05-1.78(m,2H),1.68-1.49(m,1H);LCMS (ESI): m / z(M+1) + : 479.3.

[0050] Confirmation of the configuration of the compound of formula (I) At room temperature, the compound of formula (I) (100 mg, 209.00 μmol, 1 eq) was dissolved in DCM (0.3 mL) and n-heptane (1.5 mL) was added dropwise with stirring. A clear viscous substance was observed, which did not become clear even when heated (40-50°C). When cooled to room temperature and left to stand overnight, a yellow oily substance was observed. When left to stand for 7 months at room temperature in the dark, transparent crystals were observed in the system. These crystals were collected and single crystal diffraction was measured, confirming the above configuration.

[0051] Single crystal diffraction study Equipment model: Single crystal X-ray diffraction (SC-XRD) (D8VENTURE) Conclusion: The compound of formula (I) has the molecular formula C 26 H 21 F3N4O2, monoclinic crystal system, space group C2, lattice parameters a=22.0128(6)Å, b=12.7542(3)Å, c=16.0152(4)Å, α=γ=90°, β=90.4900(10)°, volume V=4496.2(2)Å 3 and the absolute configuration parameter Flack value is 0.04(3).

[0052] Example 2: Preparation of Crystal Form A of Compound of Formula (I) At room temperature (30°C), the compound of formula (I) (79.3g, 165.74mmol, 1eq) was dissolved in ethyl acetate (80mL), and n-heptane (400mL) was slowly added dropwise. After about 100mL was added, a viscous solid was observed, so the addition rate was slowed down and the stirring rate was increased. After the addition was completed over about 2 hours, the large solid (adhering to the bottle wall) was stirred to disperse, and the mixture was further stirred at room temperature (30°C) for 16 hours. The mixture was filtered and the filter cake was collected. The filter cake was dried under reduced pressure, and the obtained filter cake was vacuum dried at 60°C overnight (16h) to obtain the crystalline form A of the compound of formula (I).

[0053] Add ethanol (60mL) to a 500mL one-neck flask containing the compound of formula (I) (30.00g, 62.70mmol, 1eq), heat to 80°C, stir until almost clear, cool to room temperature (30°C), add crystalline form A of compound of formula (I) (about 300mg), stir at room temperature (30°C) for 2 hours, add water (120mL) dropwise, and stir at room temperature (30°C) for 16 hours. Break apart large solids and stir for another 2 hours. Filter, collect the filter cake, and vacuum dry at 90°C for 32 hours to obtain crystalline form A of compound of formula (I).

[0054] Example 3: Preparation of Compound 1 [ka]

[0055] Step 1: Preparation of Compound 1-2 Preparation of LDA solution: Under nitrogen protection, n-butyllithium (2.5 M, 7.04 mL, 1.1 eq) was added dropwise to a solution of diisopropylamine (1.70 g, 16.80 mmol, 2.37 mL, 1.05 eq) in anhydrous tetrahydrofuran (30 mL) at -78°C, and the resulting mixture was warmed to 0°C and reacted for 0.5 h, then cooled to -78°C for use.

[0056] At -78°C, under nitrogen protection, the above LDA solution was dropped into anhydrous tetrahydrofuran (5mL) solution in which compound c (1.84g, 15.99mmol, 1eq) was dissolved, and the resulting mixture was reacted at -78°C for 1 hour. Anhydrous tetrahydrofuran (5mL) solution in which compound 1-1 (3.41g, 15.99mmol, 1eq) was dissolved was dropped into the reaction solution, and the resulting mixture was gradually warmed to room temperature (24°C), and the reaction was continued for another 16 hours. A saturated ammonium chloride solution was added to the system, and ethyl acetate (20mL) was added, followed by separation and extraction. The organic phase was washed with saturated saline (10mL), then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography separation to obtain compound 1-2. LCMS: MS (ESI) m / z (M-55) + :272.9.

[0057] Step 2: Preparation of Compounds 1-3 At 0°C, Dess-Martin oxidant (7.84g, 18.47mmol, 5.72mL, 1.2eq) was added to anhydrous dichloromethane (300mL) solution of compound 1-2 (5.07g, 15.44mmol, 1eq), and the temperature was gradually raised to room temperature (26°C) and reacted for 3 hours. Saturated sodium bicarbonate solution (200mL) and dichloromethane (400mL) were added to the system, filtered, the filtrate was separated, the organic phase was washed with saturated saline (100mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1-3. LCMS: MS (ESI) m / z (M-55) + :270.9.

[0058] Step 3: Preparation of Compounds 1-4 Acetic acid (5.25 g, 87.43 mmol, 5 mL, 57.06 eq) was added to a solution of compound 1-3 (500 mg, 1.53 mmol, 1 eq) and compound j (1.00 g, 4.23 mmol, 2.76 eq) in ethanol (25 mL), and the mixture was reacted at room temperature (25° C.) for 16 hours. The reaction solution was concentrated under reduced pressure to obtain compound k. LCMS: MS (ESI) m / z (M-55) + :489.1.

[0059] Step 4: Preparation of compound l Cesium carbonate (3.45 g, 10.57 mmol, 3.03 eq) was added to a solution of compound k (1.9 g, 3.49 mmol, 1 eq) in N,N-dimethylformamide (30 mL), and the mixture was heated to 135° C. and reacted for 1 hour. The reaction solution was filtered (through diatomaceous earth), the filter cake was washed with N,N-dimethylformamide (30 mL), and the filtrate was concentrated under reduced pressure and purified by column chromatography separation to obtain compound l. LCMS: MS (ESI) m / z (M+H) + :525.3.

[0060] Step 5: Preparation of Compounds 1-4 Trifluoroacetic acid (6.16 g, 54.02 mmol, 4 mL, 48.44 eq) was added dropwise to compound l (585 mg, 1.12 mmol, 1 eq) in dichloromethane (16 mL), and the mixture was allowed to react at room temperature (30° C.) for 0.5 hours. The reaction solution was concentrated under reduced pressure to obtain compound 1-4 (crude product, trifluoroacetate salt). LCMS: MS (ESI) m / z (M+1) + :425.2.

[0061] Step 6: Preparation of Compounds 1A and 1B Sodium carbonate (1.15 g, 10.85 mmol, 4.30 eq) was added to a solution of compound 1-4 (1.36 g, 2.53 mmol, 1 eq, trifluoroacetate) in tetrahydrofuran (10 mL) and water (10 mL), and a solution of acryloyl chloride (130 mg, 1.44 mmol, 117.12 μL, 5.69 e-1 eq) in tetrahydrofuran (1 mL) was added dropwise to the reaction solution, and the reaction was carried out at room temperature (25 ° C.) for 1 hour. The reaction solution was adjusted to pH=5 using 1N hydrochloric acid, extracted with dichloromethane (10mL*3), the combined organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, purified by column chromatography separation, and analyzed by supercritical fluid chromatography detection (column: Cellulose2150×4.6mmI.D, 5μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% within 5 minutes, held at 40% for 2.5 minutes, returned to 5% and equilibrated for 2.5 minutes; flow rate: 2.5mL / min; column temperature: 35℃; wavelength: 220nm), which showed that it was a racemic compound. Chiral separation (column: Phenomenex-Cellulose-2 (250 mm*30 mm, 5 μm); mobile phase: A: supercritical carbon dioxide, B: [0.1% NH3H2OETOH]; B%: 40%-40%) was performed to obtain the chiral isomers Compound 1A (retention time 6.616 min) and Compound 1B (retention time 6.971 min).

[0062] Compound 1A: 1HNMR (400 MHz, DMSO-d6) δ 9.09 (s, 1H), 8.33 (s, 1H), 7.85 (d, J = 9.0 Hz, 2H), 7.39 - 7.22 (m, 4H), 7.16 - 7.05 (m, 1H), 6.92 - 6.76 (m, 1H), 6.09 (t, J = 16.1 Hz, 1H), 5.72 - 5.57 (m, 1H), 4.72 (d, J = 12.3 Hz, 0.5H), 4.31 (t, J = 13.7 Hz, 1H), 4.17 - 4.00 (m, 0.5H), 3.53 - 3.44 (m, 0.5H), 3.33 - 3.14 (m, 1.5H), 3.12 - 3.02 (m, 0.5H), 3.01 - 2.89 (m, 0.5H), 2.27 - 2.16 (m, 1H), 2.05 - 1.80 (m, 2H), 1.66 - 1.46 (m, 1H). LCMS: MS (ESI) m / z (M+1) + : 479.2。

[0063] Compound 1B: 1 HNMR (400 MHz, DMSO-d6) δ 9.09 (s, 1H), 8.33 (s, 1H), 7.85 (d, J = 8.8 Hz, 2H), 7.40 - 7.23 (m, 4H), 7.14 - 7.04 (m, 1H), 6.94 - 6.75 (m, 1H), 6.10 (t, J = 16.1 Hz, 1H), 5.72 - 5.52 (m, 1H), 4.73 (d, J = 12.5 Hz, 0.5H), 4.32 (t, J = 12.7 Hz, 1H), 4.09 (d, J = 13.1 Hz, 0.5H), 3.57 - 3.44 (m, 0.5H), 3.33 - 3.15 (m, 1.5H), 3.12 - 3.03 (m, 0.5H), 3.03 - 2.88 (m, 0.5H), 2.27 - 2.16 (m, 1H), 2.05 - 1.79 (m, 2H), 1.68 - 1.49 (m, 1H). LCMS: MS (ESI) m / z (M+1) + : 479.2。

[0064] (Example 4) Examination of the hygroscopicity of crystalline form A of the compound of formula (I) Experimental materials SMSDVS IntrinsicPlus dynamic vapor sorption apparatus Experimental method Samples (10 - 20 mg) were placed in a DVS sample tray and tested. Experimental Results The DVS spectrum of crystalline form A of compound of formula (I) is shown in FIG. 4, and ΔW=0.23%. Experimental Conclusion At 80% RH / 25° C., the crystalline form A of compound of formula (I) exhibited a moisture weight gain of 0.23%, indicating that the sample is slightly hygroscopic.

[0065] Example 5: Testing the Solid State Stability of Crystalline Form A of Compound of Formula (I) According to the Guidance Principles for Stability Testing of Drug Substances and Preparations (Chinese Pharmacopoeia 2015, Part 4 General Principles 9001), the influencing factors of crystalline form A of compound of formula (I) were ((high temperature (60°C, open), high humidity (room temperature / relative humidity 92.5%, open) and strong light irradiation (visible light intensity 5000 lux and ultraviolet intensity 90 w / cm 2 The stability under accelerated conditions ((40°C / 75% RH, open) and (60°C / 75% RH, open)) was examined.

[0066] Approximately 20 mg of crystalline form A of compound of formula (I) was weighed out and placed on the bottom of a glass sample bottle, spread thinly, and left to stand with the sample fully exposed. The samples left under different conditions were sampled and detected (XRPD) on the 5th day, 10th day, 1st month, 2nd month, and 3rd month, and the detection results were compared with the initial detection results on the 0th day, and the test results are shown in Table 3 below. [Table 3] Conclusion: The crystalline form A of compound of formula (I) has excellent stability under both influencing factors and accelerated conditions.

[0067] Biological Test Data: Experimental Example 1: EGFR, ITK, TEC, and BTK Kinase Assay 1.Reaction conditions Buffer conditions: 20mMHEPES (pH7.5), 10mMgCl2, 1mMEGTA, 0.02%Brij35, 0.02mg / mLBSA, 0.1mmNa3VO4, 2mmDTT, 1%DMSO.

[0068] 2. Reaction Procedure 2.1. Prepare indicator substrate in freshly prepared reaction buffer. 2.2. The desired cofactor was transferred to the substrate solution above. 2.3. The indicated kinase was added to the substrate solution and mixed gently. 2.4. Compounds in DMSO were delivered to the kinase reaction mixture (Echo550) using acoustic techniques. 2.5. 33 P-ATP (specific activity 0.01 μci / μL final) was added to the reaction mixture to initiate the reaction (final ATP concentrations were 2 μM, 5 μM, and 5 μM, respectively). 2.6. The kinase reaction was incubated at room temperature for 120 minutes. 2.7. The reaction was recorded on P81 ion exchange paper (Whatman #3698-915). 2.8.0.75% phosphoric acid cleaning filters were used extensively. 2.9. The amount of radioactive phosphorylated substrate remaining on the filter paper was measured.

[0069] 3. Data Analysis: Kinase activity data are expressed as the percentage of remaining kinase activity relative to reaction with carrier (dimethyl sulfoxide) in the test sample, IC 50 Values ​​and curve fitting were obtained with Prism4 software (GraphPad).

[0070] 4. Experimental conclusion: The results are shown in Table 4. [Table 4] Conclusion: The compounds of the present invention have excellent EGFR, ITK, and TEC kinase selectivity.

Claims

1. Crystal A of compound of formula (I), Crystal A of compound of formula (I) is characterized in that it has characteristic diffraction peaks at 2θ angles of 17.7805±0.2000°, 22.0193±0.2000°, and 27.4192±0.20° in its powder X-ray diffraction pattern. 【Chemistry 1】

2. The crystal A according to claim 1, having characteristic diffraction peaks at 2θ angles of 15.0010±0.2000°, 17.0603±0.2000°, 17.7805±0.2000°, 22.0193±0.2000°, 23.5013±0.2000°, and 27.4192±0.2000° in a powder X-ray diffraction pattern.

3. The crystalline A according to claim 2, having characteristic diffraction peaks at 2θ angles of 8.1387±0.2000°, 15.0010±0.2000°, 16.1591±0.2000°, 17.0603±0.2000°, 17.7805±0.2000°, 22.0193±0.2000°, 23.5013±0.2000°, and 27.4192±0.2000° in a powder X-ray diffraction pattern.

4. The crystalline A according to claim 3, having characteristic diffraction peaks at 2θ angles of 8.1387±0.2000°, 13.6809±0.2000°, 15.0010±0.2000°, 16.1591±0.2000°, 17.0603±0.2000°, 17.7805±0.2000°, 22.0193±0.2000°, 23.5013±0.2000°, 24.0218±0.2000°, and 27.4192±0.2000° in a powder X-ray diffraction pattern.

5. In the powder X-ray diffraction pattern, the following are observed: 8.1387°, 9.8194°, 13.6809°, 15.0010°, 16.1591°, 17.0603°, 17.7805°, 18.4386°, 19.5400°, 21.3193°, 22.0193°, 23.1597°, 23.5013°, 24.0218°, 26.1791°, The crystal A according to claim 4, having characteristic diffraction peaks at 2θ angles of 26.6006°, 27.1199°, 27.4192°, 29.1595°, 29.7190°, 30.8417°, 31.2196°, 32.2992°, 32.9612°, 33.7773°, 34.3779°, 35.1796°, and 37.1408°.

6. The crystal A according to any one of claims 1 to 5, wherein the differential scanning calorimetry curve has an endothermic peak at 107.89 ± 3.00 ° C. and an exothermic peak at 147.76 ± 3.00 ° C.

7. The crystal A according to any one of claims 1 to 5, wherein the weight loss at 200.0±3°C in the thermogravimetric analysis curve reaches 0.06%.

8. Its single crystal X-ray diffraction data shows a monoclinic system, space group C2, with lattice parameters a = 22.0128(6) Å, b = 12.7542(3) Å, c = 16.0152(4) Å, α = γ = 90°, β = 90.4900(10)°, and volume V = 4496.2(2) Å. ３ The crystal A according to claim 1, having an absolute configuration parameter Flack value of 0.04 (3).

9. A method for preparing Crystal A of compound of formula (I), comprising the steps of: Step 1) adding a compound of formula (I) to a solvent to form a solution; 2) adding an anti-solvent to the solution, stirring at a predetermined temperature for a predetermined time, filtering, and drying the filter cake under reduced pressure; the solvent is an ester-based solvent, an ether-based solvent, or an alcohol-based solvent, the anti-solvent is n-heptane, n-hexane or water; The stirring temperature is 0 to 40° C. The method for producing Crystal A of compound of formula (I), wherein the stirring time is 12 to 48 hours.

10. Use of Crystal A according to any one of claims 1 to 8 in the manufacture of a drug for treating a BTK-related disease.

11. The use of claim 10, wherein the BTK-related disease is a hematological tumor or an autoimmune disease.

Images