A salt type of a spiro compound, a crystal form thereof, and a preparation method thereof

Abstract

Disclosed are a salt type and crystal form of a spiro compound, a preparation method of the salt type and crystal form, and application of the salt type and crystal form in drugs for treating related diseases.

Description

Technical Field

[0001] This invention discloses a salt form and crystal form of a spirocyclic compound and a method for preparing the same, as well as the application of the salt form and crystal form in drugs for related diseases. Background Technology

[0002] Non-alcoholic fatty liver disease (NAFLD) has a high prevalence in developed countries and regions, ranging from approximately 15% to 40%. Of these, 10% to 20% of NAFLD patients will develop non-alcoholic steatohepatitis (NASH). The estimated global incidence of NASH is 5% to 7%, which increases to 22% in diabetic patients. Notably, approximately 15% to 25% of NASH patients will develop cirrhosis.

[0003] Recent studies have found that a high-fructose diet is a significant cause of NASH. Once in the liver, fructose is rapidly phosphorylated into fructose-1-phosphate by ketohexokinase (KHK). Further metabolites produced by fructose-1-phosphate become substrates for gluconeogenesis and de novo lipid synthesis (DNL), leading to increased hepatic lipid synthesis and insulin resistance, thereby increasing oxidative stress and inflammation, and accelerating the pathogenesis of NAFLD and NASH. KHK is the rate-limiting enzyme in the metabolism of fructose to fructose-1-phosphate and is an important target for regulating fructose metabolism. Therefore, inhibiting KHK can effectively suppress fructose metabolism and its resulting lipid accumulation, oxidative stress, inflammation, and insulin resistance, thus potentially contributing to the treatment of NASH. Summary of the Invention

[0004] This invention provides compounds of formula (II), .

[0005] The present invention also provides a crystal form A of the compound of formula (II), characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 9.73±0.20°, 11.22±0.20°, 12.66±0.20°, 18.38±0.20°.

[0006] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 6.94±0.20°, 9.73±0.20°, 11.22±0.20°, 12.66±0.20°, 15.63±0.20°, 16.62±0.20°, 18.38±0.20°, 20.81±0.20°.

[0007] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 6.46±0.20°, 6.94±0.20°, 9.73±0.20°, 11.22±0.20°, 12.66±0.20°, 15.63±0.20°, 16.62±0.20°, 18.38±0.20°, 19.64±0.20°, 20.81±0.20°, 22.51±0.20°, 24.58±0.20°.

[0008] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 4.47±0.20°, 6.46±0.20°, 6.94±0.20°, 9.73±0.20°, 11.22±0.20°, 12.25±0.20°, 12.66±0.20°, 15.63±0.20°, 16.62±0.20°, 17.83±0.20°, 18.38±0.20°, 19.64±0.20°, 20.81±0.20°, 22.51±0.20°, 23.59±0.20°, 24.58±0.20°.

[0009] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned A-type crystal has characteristic diffraction peaks at the following 2θ angles: 4.47°, 6.46°, 6.94°, 9.73°, 10.18°, 11.22°, 12.25°, 12.66°, 13.30°, 14.30°, 15.63°, 16.62°, 17.27°, 17.83°, 18.38°, 18.67°, 19.37°, 19.64°, 20.06°, 20.41°, 20.81°, 21.72°, 22.51°, 23.59°, 24.58°, 25.11°, 26.44°, 27.41°, 27.99°, 29.40°, and 30.85°.

[0010] In some embodiments of the present invention, the XRPD pattern of the above-mentioned A-type crystal is as follows: Figure 1 As shown.

[0011] In some embodiments of the present invention, the XRPD spectra analysis data of the above-mentioned A-type crystal form are shown in Table 1: Table 1

[0012] In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned A crystal form has a peak value of an endothermic peak at 188.5℃±3.0℃.

[0013] In some embodiments of the present invention, the DSC spectrum of the above-mentioned A-type crystal is as follows: Figure 2 As shown.

[0014] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned A crystal form shows a weight loss of 1.56% at 150℃±3℃.

[0015] In some embodiments of the present invention, the TGA pattern of the above-mentioned A-type crystal is as follows: Figure 3 As shown.

[0016] The present invention also provides a method for preparing crystal form A of compound (II), comprising: (a) Dissolve the compound of formula (I) in an alcohol solvent; (b) Slowly add a mixture of betaine and alcohol solvent while stirring; (c) Stir at 20~30℃ for 8~16 hours; (d) After filtration, dry for 8-16 hours; The alcohol solvent is selected from isopropanol, ethanol and methanol.

[0017] The present invention also provides the use of the compound of formula (II) or its A crystal form or the crystal form prepared according to the above method in the preparation of a medicament for treating non-alcoholic steatohepatitis (NASH).

[0018] Beneficial effects The betaine salt form described in this invention exhibits better stability (including hygroscopic stability and storage stability) compared to other salt forms. For example, it is stable for one month at 40°C / 75% RH without the detection of byproducts. The betaine salt form also possesses better crystallinity and higher purity. Furthermore, the compounds described in this invention have stable crystal forms and good drug-like properties.

[0019] Definitions and Explanations Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular phrase or term should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When trade names appear herein, they are intended to refer to the corresponding product or its active ingredient.

[0020] 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 equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0021] The chemical reactions in the specific embodiments of this invention are carried out in a suitable solvent, which must be suitable for the chemical changes of this invention and the reagents and materials required therefor. To obtain the compounds of this invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0022] The structures of the compounds of this invention can be confirmed using conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, that absolute configuration can be confirmed using conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) is used, where the cultured single crystal is used to collect diffraction intensity data using a Bruker D8 venture diffractometer with CuKα radiation as the light source. The scanning method is as follows: After collecting relevant data by ω scanning, the crystal structure can be further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0023] The present invention will be described in detail below through embodiments, which are not intended to limit the present invention in any way.

[0024] All solvents used in this invention are commercially available and can be used without further purification.

[0025] The following abbreviations are used in this invention: rt represents room temperature; THF represents tetrahydrofuran; NMP represents N-methylpyrrolidone; MeSO3H represents methanesulfonic acid; DME represents ethylene glycol dimethyl ether; DCM represents dichloromethane; Xphos represents 2-bicyclohexylphosphine-2'4'6'-triisopropylbiphenyl; EtOAc represents ethyl acetate; MeOH represents methanol; acetone represents acetone; 2-Me-THF represents 2-methyltetrahydrofuran; IPA represents isopropanol; DAST represents diethylaminotrifluoride.

[0026] Compounds are named manually or using ChemDraw® software; commercially available compounds are named using supplier catalog names.

[0027] The present invention relates to a powder X-ray diffractometer (XRPD) method. Instrument Model: Bruker D8 Advance X-ray Diffractometer Test method: Approximately 10 ~ 20 mg of sample is used for XRPD detection.

[0028] The detailed XRPD parameters are as follows: Optical tube: Cu, kα, (λ=1.54056) ). Phototube voltage: 40 kV, Phototube current: 40 mA Diverging slit: 0.60 mm Detector slit: 10.50 mm Anti-scattering slit: 7.10 mm Scan range: 4-40 deg Step length: 0.02 deg Step length: 0.12 seconds Sample tray rotation speed: 15 rpm This invention relates to a differential scanning calorimeter (DSC) method. Instrument Model: TA Q2000 Differential Scanning Calorimeter Test method: Take a sample (~1 mg) and place it in a DSC aluminum pot for testing. Under N2 conditions of 50 mL / min, heat the sample from 30℃ (room temperature) to 300℃ (or 350℃) at a heating rate of 10℃ / min.

[0029] The present invention relates to a thermogravimetric analysis (TGA) method. Instrument Model: TA Q5000IR Thermogravimetric Analyzer Test method: Take a sample (2~5mg) and place it in a TGA platinum pot for testing. Under N2 conditions of 25mL / min, heat the sample from room temperature to 350℃ or lose 20% of its weight at a heating rate of 10℃ / min.

[0030] This invention presents a method for dynamic vapor adsorption analysis (DVS). Instrument Model: SMS DVS Advantage Dynamic Vapor Adsorption Analyzer Test conditions: Take a sample (10 ~ 15 mg) and place it in the DVS sample tray for testing.

[0031] The detailed DVS parameters are as follows: Temperature: 25℃ Equilibrium: dm / dt = 0.01 % / min (shortest: 10 min, longest: 180 min) Drying: Dry at 0% RH for 120 min RH (%) test tier: 10% RH (%) test range: 0% - 90% - 0% Hygroscopicity evaluation is classified as follows:

[0032] Note: ΔW% represents the moisture gain of the test sample at 25 ± 1℃ and 80 ± 2% RH. Attached Figure Description

[0033] Figure 1 XRPD spectrum of Cu-Kα radiation for crystal form A of compound (II); Figure 2 The DSC spectrum of compound A (II) is shown below. Figure 3 The TGA spectrum of compound A (II) is shown below. Figure 4 The DVS spectrum of compound A (II) is shown. Detailed Implementation

[0034] To better understand the content of this invention, further explanation will be provided below with reference to specific embodiments. However, the specific implementation methods are not intended to limit the content of this invention.

[0035] Example 1: Preparation of compound (I) Synthesis route: Step 1: Synthesis of compound A-1_2 A-1_1 (50 g, 264.49 mmol) and NaOMe (100 g, 1.85 mol) were dissolved in MeOH (500 mL), and the reaction was carried out under nitrogen protection at 80 °C with stirring for 12 hours. LC-MS showed that the starting material signal disappeared and the product signal was formed. TLC (petroleum ether: ethyl acetate = 3:1) showed the formation of new spots. The reaction solution was directly evaporated to dryness, water (500 mL) was added, and the mixture was extracted with EtOAc (400 mL). The organic phase was evaporated to dryness to obtain A-1_2.

[0036] Step 2: Synthesis of compound A-1_3 A-1_2 (90 g, 499.44 mmol) and CHCl3 (1000 mL) were added to a 3000 mL three-necked flask, followed by the addition of m-chloroperoxybenzoic acid (287.23 g, 1.41 mmol, 85% purity). The reaction was carried out under nitrogen protection and stirred at 30 °C for 12 hours. LCMS showed that the starting material signal did not disappear and a product signal was generated. TLC (dichloromethane:methanol = 10:1) showed the formation of new spots. The reaction solution was filtered, and the filter cake was washed with dichloromethane (500 mL). The filtrate was slowly added to a saturated sodium sulfite solution (500 g of sodium sulfite was used to prepare approximately 2.5 L). The solution was stirred for one hour to quench the oxidizing agent. The mixture was separated, and the aqueous phase was washed with 1000 mL of dichloromethane. The organic phases were combined and evaporated to dryness. 1000 mL of methyl tert-butyl ether was added, and the organic phase was washed with a saturated sodium carbonate solution (500 mL × 3). The aqueous phases were combined, and the aqueous phase was washed again with 500 mL of methyl tert-butyl ether (sodium carbonate solution). The aqueous phases were combined and extracted with chloroform (2 L × 4). The chloroform and organic phases were combined and evaporated to dryness to obtain A-1_3. 1 H NMR(400MHz, CDCl3)δ = 4.24 - 4.11(m, 3H), 4.06 - 3.95(m, 3H), 3.20(t, J=7.8 Hz, 2H), 2.86(t, J=7.7 Hz, 2H), 2.28- 2.15(m, 2H).

[0037] Step 3: Synthesis of compound A-1_4 A-1_3 (59 g, 300.71 mmol) was added to a 1000 mL single-necked flask, followed by acetic anhydride (250 mL). The reaction mixture was stirred at 80 °C for 5 hours under nitrogen protection. The reaction solution was slowly added to water (500 mL), and extracted with ethyl acetate (300 mL × 2). The organic phase was directly evaporated to dryness to obtain the crude product. The crude product was purified by rapid silica gel column chromatography (ISCO cake, 330 g SepaFlash silica gel column, eluent: 0–10% EtOAc / PE, flow rate 100 mL / min) to obtain A-1_4. 1 H NMR(400 MHz, CDCl3)δ = 6.12 - 5.90(m, 1H), 4.02(d, J=7.0 Hz, 6H), 2.95 - 2.82(m,1H), 2.78 - 2.57(m, 2H), 2.14(s, 3H), 2.08 - 1.96(m, 1H).

[0038] Step 4: Synthesis of compound A-1_5 A-1_4 (40 g, 167.9 mmol), THF (400 mL) / H2O (100 mL) were added to a 1000 mL single-necked flask, followed by LiOH·H2O (14 g, 335.8 mmol). The reaction mixture was stirred at 20 °C for 12 hours under nitrogen protection. The reaction solution was then evaporated to dryness. The crude product was purified by rapid silica gel column chromatography (ISCO 330 g SepaFlash silica gel column, eluent: 0–20% EtOAc / PE, flow rate 35 mL / min) to obtain A-1_5. 1 H NMR(400MHz, CDCl3)δ= 5.10(t, J=7.0 Hz, 1H),4.10 - 3.96(m, 6H), 2.88(ddd, J=2.8, 8.9, 15.4 Hz, 1H), 2.70 - 2.48(m, 2H),2.12 - 1.94(m, 1H).

[0039] Step 5: Synthesis of compound A-1_6 A-1_5 (150 g, 764.52 mmol) and DCM (1500 mL) were added to a 5 L three-necked flask, along with Dysmart oxidant (660 g, 1.56 mol). The reaction mixture was stirred at 20 °C for 12 hours under nitrogen protection. The reaction mixture was filtered directly, and the filter cake was washed with ethyl acetate (200 mL). The filtrate was then evaporated to dryness. A-1_6 was purified using a rapid silica gel column (ISCO cake, 330 g SepaFlash rapid silica gel column, eluent: 0–10% EtOAc / PE, flow rate 100 mL / min) to obtain A-1_6. 1 HNMR(400MHz, CDCl3)δ = 4.02(d, J=8.3 Hz, 6H), 2.92 - 2.82(m, 2H), 2.71 - 2.62(m, 2H).

[0040] Step 6: Synthesis of compound A-1_7 A-1_6 (50 g, 257.48 mmol) and DCM (500 mL) were added to a 1000 mL single-necked flask, followed by DAST (122 g, 756.88 mmol, 100 mL). The reaction mixture was stirred at 30 °C for 20 hours under nitrogen protection. The reaction mixture was slowly quenched by adding ice water (2000 mL), and the filter cake was washed with dichloromethane (2000 mL). The filtrate was then evaporated to dryness. A-1_7 was obtained by purification using a rapid silica gel column (ISCO cake, 330 g SepaFlash silica gel column, eluent: 0–10% EtOAc / PE, flow rate 100 mL / min).1 H NMR(400MHz, CDCl3)δ = 3.98(d, J=5.1 Hz, 6H), 2.83 - 2.70(m,2H), 2.62 - 2.41(m, 2H).

[0041] Step 7: Synthesis of compound A-1_8 A-1_7 (50 g, 231.28 mmol) and THF (100 mL) were added to a 1000 mL single-necked flask, and concentrated hydrochloric acid (500 mL) was added. The reaction was carried out under nitrogen protection and stirred at 80 °C for 12 hours. The reaction solution was slowly cooled to room temperature, the turbid liquid was filtered, and the filter cake was washed with ethyl acetate (50 mL) to obtain A-1_8. 1 H NMR(400MHz, CDCl3)δ = 11.85(brs, 1H), 11.36 - 11.12(m, 1H), 2.61 - 2.52(m, 4H).

[0042] Step 8: Synthesis of compound A-1 A-1_8 (34 g, 180.72 mmol) was added to a 1000 mL single-necked flask, followed by POCl3 (206 mL). The reaction mixture was stirred at 120 °C for 12 hours under nitrogen protection. The reaction solution was evaporated to dryness, diluted with dichloromethane (500 mL), and then slowly added to water (1500 mL) to quench the reaction. The organic phase was then extracted with dichloromethane (1000 mL × 3), and the solutions were evaporated to dryness to obtain A-1. 1 HNMR(400MHz, CDCl3)δ = 3.16 - 3.01(m, 2H), 2.85 - 2.65(m, 2H).

[0043] Step 9: Synthesis of compound a Compound B-1 (40 g, 129.28 mmol) was dissolved in DCM (300.0 mL), and the solution was cooled to 0 °C. A solution of compound A-1 (27 g, 119.99 mmol) in DCM (200.0 mL) was slowly added dropwise, followed by the slow addition of DIPEA (46.52 g, 359.97 mmol). The reaction was stirred at 0 °C for 3 hours. After the reaction was complete, the solvent was removed by rotary evaporation to obtain the crude product. The crude product was purified by automated column chromatography (100–200 mesh, eluent: PE:EtOAc = 100:1–100:50) to obtain a. 1H NMR(400MHz, CDCl3) δ = 4.73 - 4.23(m, 4H), 3.72(d, J=5.5 Hz, 3H), 3.07 - 2.90(m, 2H), 2.67 - 2.48(m, 2H), 1.93(br d, J=9.5 Hz, 1H), 1.72 - 1.51(m, 1H), 1.75 - 1.49(m, 1H).

[0044] Step 10: Synthesis of compound b Compound a (35 g, 106.15 mmol) was added in portions to a solution of C-1 hydrochloride (38.65 g, 127.38 mmol) in acetonitrile (350.0 mL), followed by the addition of K₂CO₃ (44 g, 318.44 mmol). The reaction was stirred at 80 °C for 12 hours. After the reaction was complete, the mixture was filtered directly, and the filtrate was evaporated to dryness to obtain compound b. 1 H NMR(400MHz, CDCl3)δ = 4.49 - 4.35(m,2H), 4.33 - 4.17(m, 3H), 4.07(dt, J=5.0, 8.8 Hz, 1H), 3.74(s, 3H), 2.87 -2.77(m, 2H), 2.53 - 2.36(m, 3H), 1.97 - 1.86(m, 2H), 1.50(d, J=6.0 Hz, 3H), 1.39(t, J=5.5 Hz, 1H), 1.32 - 1.26(m, 1H), 1.30 - 1.24(m, 1H).

[0045] Step 11: Synthesis of Compound I Compound b (36 g, 98.8 mmol) was dissolved in THF (350.0 mL) and H₂O (70.0 mL), and lithium hydroxide monohydrate (8.29 g, 197.59 mmol) was added. The reaction was stirred at 20 °C for 12 hours. After the reaction was complete, 200 mL of water was added, and the pH was adjusted to 5-6 with 1N hydrochloric acid. Extraction was performed with ethyl acetate (300 mL). The solvent was removed by rotary evaporation, and the product was dissolved in 25 mL of MeOH. Insoluble inorganic salts were removed by filtration. The MeOH solution of the crude product was purified by Prep-HPLC (separation method: column type: Phenomenex Luna C8 250*50mm*10μm; mobile phase: [H₂O (0.225% formic acid)-ACN]; ACN%: 5%-35%, 10 min) to obtain compound I. 1H NMR(400 MHz, CD3OD)δ ppm 4.22 - 4.50(m, 5H),3.86 - 4.12(m, 2H), 2.88(br d, J =3.76 Hz, 2H), 2.32 - 2.55(m, 3H), 1.84 - 2.04(m, 2H), 1.49(dd, J =6.02, 1.51 Hz, 3H), 1.22 - 1.37(m, 2H). MS (ESI): m / z:351.1 [M+1].

[0046] Example 2: Preparation of crystal form A of compound (II) 20 mL of isopropanol was added to a 100 mL reaction flask. Compound (I) (2.0 g, 5.71 mmol, 1 eq) was added to the reaction flask at room temperature and stirred until dissolved and clear. A supernatant solution of betaine (702.15 mg, 5.99 mmol, 1.05 eq) and MeOH (5 mL) was added to the above reaction solution, and the mixture was stirred overnight at 25°C. The mixture was filtered under reduced pressure, and the filter cake was washed with isopropanol (5 mL). The filter cake was then dried under vacuum to constant weight to obtain crystal form A of compound (II).

[0047] 1H NMR (400 MHz, CD3OD): δ, 4.45-4.35 (m, 5 H), 4.04-4.02 (m, 1 H) ,3.93-3.91 (m, 1 H) 3.85 (s, 2 H), 3.29 (s, 9 H) ,2.88-2.86(m, 2 H) ,2.49–2.40(m, 3 H) , 1.95–1.89 (m, 2 H), 1.49 (d, J = 6.0 Hz,3 H), 1.32 (t, J = 4.0 Hz,2 H). Example 3: Study on the hygroscopicity of crystal form A of compound (II) Experimental materials: SMS DVS Advantage Dynamic Vapor Adsorption Analyzer Experimental methods: Take 10-15 mg of compound A of formula (II) and place it in the DVS sample tray for testing.

[0048] Experimental results: The DVS spectrum of compound A of formula (II) is as follows: Figure 3 As shown, △W = 0.29%.

[0049] Experimental conclusion: The A crystal form of compound (II) has a moisture absorption weight gain of 0.29% at 25°C and 80% RH, indicating slight hygroscopicity.

[0050] Biological test data Experimental Example 1: Fructose-Kinetic Kinase Assay (KHK assay) A. Main materials 1. EnVision microplate reader, PerkinElmer; 2. OptiPlate 384 microplate, PerkinElmer, part number: 6007290; 3. Recombinant human fructokinase (KHK), R&D catalog number: 8177-HK-020, batch number: DDFK0117092; 4. Fructose (D(-)-Fructose), National Pharmaceutical Group Co., Ltd. Item No.: 36003034; 5. ADP-Glo ​​reagent kit, Promega, catalog number: V9101.

[0051] B. Method a) Kinase response 1. Prepare the buffer solution: containing 50 mM hydroxyethylpiperazine ethanethioic acid (Hepes), 140 mM KCl, 3.5 mM MgCl2, 0.01% bovine serum albumin (BSA), with a pH of 7.4.

[0052] 2. Prepare a 2.5-fold concentration of fructokinase working solution using buffer, wherein the fructokinase is 50 nM and the fructose is 12.5 mM.

[0053] 3. Prepare a 2.5-fold concentration of adenosine triphosphate (ATP) working solution using buffer solution, with a concentration of 250 μM.

[0054] 4. Dilute the compound starting at a concentration of 500 µM, and dilute it 3 times at 9 concentration points. The final concentration of the compound in the reaction system starts at 10 µM, and the final concentration of dimethyl sulfoxide (DMSO) is 2%.

[0055] 5. Prepare a 96-well plate as a reaction plate, add 6 μL of 2.5 times the concentration of fructokinase working solution to each well, then add 3 μL of compound working solution to each well, and incubate at room temperature for 5 minutes.

[0056] 6. The first well in each row is the positive control for the compound, i.e., the same volume of buffer solution is added to replace the compound and fructokinase; the last well is the negative control for the compound, i.e., the same volume of buffer solution is added to replace the compound.

[0057] 7. After adding 6 μL of ATP working solution to each well of a 96-well plate, the kinase reaction was initiated. The kinase reaction was incubated in a constant temperature heater at 28°C for 1 hour.

[0058] b) ADP-Glo ​​detection 1. Prepare a 384 plate as a detection plate and add 5 μL of ADP-Glo ​​reagent.

[0059] 2. Add 5 μL of the kinase reaction mixture from the reaction plate to each well and incubate in a constant temperature heater at 28°C for 30 minutes.

[0060] 3. Add 10 μL of kinase detection reagent to each well and incubate in a constant temperature heater at 28°C for 30 minutes.

[0061] 4. Place the test plate into the EnVision microplate reader to read the chemiluminescence signal.

[0062] C. Experimental Results: Table 2: Results of KHK in vitro activity test

[0063] Conclusion: Compound I exhibits strong inhibitory activity against human KHK enzyme.

Claims

1. Compound of formula (II), 。 2. Crystal form A of compound (II): ， Its features Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 9.73±0.20°, 11.22±0.20°, 12.66±0.20°, 16.62±0.20°, and 18.38±0.20°.

3. The A-type crystal according to claim 2, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.94±0.20°, 9.73±0.20°, 11.22±0.20°, 12.66±0.20°, 15.63±0.20°, 16.62±0.20°, 18.38±0.20°, 20.81±0.20°.

4. The A-type crystal according to claim 3, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.46±0.20°, 6.94±0.20°, 9.73±0.20°, 11.22±0.20°, 12.66±0.20°, 15.63±0.20°, 16.62±0.20°, 18.38±0.20°, 19.64±0.20°, 20.81±0.20°, 22.51±0.20°, 24.58±0.20°.

5. The A-type crystal according to claim 4, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.47±0.20°, 6.46±0.20°, 6.94±0.20°, 9.73±0.20°, 11.22±0.20°, 12.25±0.20°, 12.66±0.20°, 15.63±0.20°, 16.62±0.20°, 17.83±0.20°, 18.38±0.20°, 19.64±0.20°, 20.81±0.20°, 22.51±0.20°, 23.59±0.20°, 24.58±0.20°.

6. The A-type crystal according to claim 5, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 4.47°, 6.46°, 6.94°, 9.73°, 10.18°, 11.22°, 12.25°, 12.66°, 13.30°, 14.30°, 15.63°, 16.62°, 17.27°, 17.83°, 18.38°, 18.67°, 19.37°, 19.64°, 20.06°, 20.41°, 20.81°, 21.72°, 22.51°, 23.59°, 24.58°, 25.11°, 26.44°, 27.41°, 27.99°, 29.40°, 30.85°.

7. The A crystal form according to any one of claims 2 to 6, wherein the differential scanning calorimetry curve has a peak value of an endothermic peak at 188.5℃±3.0℃.

8. The A-type crystal according to claim 7, its DSC spectrum is shown in Figure 2.

9. The A crystal form according to any one of claims 2 to 6, wherein the thermogravimetric analysis curve shows a weight loss of 1.56% at 150.0℃ ± 3.0℃.

10. The A crystal form according to claim 9, its TGA spectrum is shown in Figure 3.

11. A method for preparing crystal form A of compound (II) as described in any one of claims 2-10, the method comprising: (a) Dissolve the compound of formula (I) in an alcohol solvent; (b) Slowly add a mixture of betaine and alcohol solvent while stirring; (c) Stir at 20~30℃ for 8~16 hours; (d) After filtration, dry for 8-16 hours; The alcohol solvent is selected from isopropanol, ethanol and methanol.

12. The use of the compound according to claim 1, or the A crystal form according to any one of claims 2 to 10, or the crystal form prepared by the method according to claim 11 in the preparation of a medicament for treating non-alcoholic steatohepatitis (NASH).

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