Polymorph of NLRP3 inhibitor, preparation method therefor, and use thereof

Abstract

Disclosed in the present invention are a polymorph of an NLRP3 inhibitor, a preparation method therefor, and use thereof. A polymorph of a compound of formula (I), comprising an amorphous form, crystal form I, crystal form II, crystal form III, and crystal form IV of the compound of formula (I). The polymorph has good hygroscopicity, solubility, physical stability, and / or chemical stability, and thus is suitable for drug development.

Description

Polymorphs of NLRP3 inhibitors, their preparation methods and applications

[0001] This application claims priority to an earlier application filed on June 28, 2024, with the China National Intellectual Property Administration, patent application number 2024108647047, entitled "Polymorphs of NLRP3 Inhibitors and Their Preparation Methods and Applications"; the entire contents of the earlier application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of compounds, specifically relating to polymorphs of NLRP3 inhibitors, their preparation methods, and applications. Background Technology

[0003] PCT / CN2023 / 142422 (application date December 27, 2023) describes the compound (S)-3-((4-(4-chloro-2-hydroxyphenyl)-7-(prop-1-yn-1-yl)phthalazin-1-yl)amino)propane-1,2-diol, the structure of which is shown in Formula (I). This compound exhibits good NLRP3 inhibitory activity, along with favorable metabolic properties and safety. Animal studies have shown that this compound has significant IL-1β inhibitory activity and holds promise for the treatment of various diseases, including gouty arthritis, demonstrating potential drug development potential.

[0004] Drugs typically exist in various solid forms, and researching suitable solid forms for drug development is a technical problem that those skilled in the art are dedicated to solving. Summary of the Invention

[0005] To improve the above-mentioned technical problems, the present invention provides polymorphs of the compound of formula (I), including crystal form I, crystal form II, crystal form III, crystal form IV and crystal form V of the compound of formula (I);

[0006] According to an embodiment of the present invention, the crystal form I, using Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 17.2±0.20°, 22.0±0.20°, 22.6±0.20°, 24.9±0.20°, 25.7±0.20°, and 27.4±0.20° when expressed in 2θ angles; further, it may also exhibit characteristic peaks at 8.6±0.20°, 10.9±0.20°, 14.4±0.20°, 20.6±0.20°, and / or 29.7±0.20°; even further, it may also exhibit characteristic peaks at 9.5±0.20°, 18.6±0.20°, 20.0±0.20°, 23.8±0.20°, 31.7±0.20°, and / or 32.7±0.20°.

[0007] In one embodiment, crystal form I is subjected to Cu-Kα radiation, and X-ray powder diffraction, expressed in 2θ angles, at 3.0±0.20°, 3.3±0.20°, 4.2±0.20°, 4.5±0.20°, 5.1±0.20°, 6.1±0.20°, 6.4±0.20°, 6.7±0.20°, 8.6±0.20°, 9.5±0.20°, 9.9±0.20°, 10.9±0.20°, 11.4±0.20°, 12.6±0.20°, 13.3±0.20°, 14.4±0.20°, and 15.1°. Characteristic peaks are observed at ±0.20°, 15.4±0.20°, 15.9±0.20°, 17.2±0.20°, 18.6±0.20°, 19.1±0.20°, 20.0±0.20°, 20.6±0.20°, 22.0±0.20°, 22.6±0.20°, 23.8±0.20°, 24.9±0.20°, 25.7±0.20°, 27.4±0.20°, 29.7±0.20°, 31.2±0.20°, 31.7±0.20°, 32.2±0.20°, and 32.7±0.20°.

[0008] According to an embodiment of the present invention, the crystal form I has an XRPD pattern as shown in FIG1.

[0009] According to an embodiment of the present invention, the crystal form I loses no more than 4 wt% of weight at room temperature to about 160°C, preferably no more than 3 wt%, for example, 2.7 wt%.

[0010] According to an embodiment of the present invention, the crystal form I is a hydrate, for example, a 0.5 to 2 hydrate.

[0011] According to an exemplary embodiment of the present invention, the crystal form I has a TGA pattern as shown in FIG2.

[0012] According to an embodiment of the present invention, crystal form I has one, two, or three peaks as shown in (a)-(c):

[0013] (a) An endothermic peak with an initial temperature of 25–30 °C and a peak temperature difference of 55–65 °C from the initial temperature; for example, crystal form I has an endothermic peak with an initial temperature of 27.66 °C and a peak temperature difference of 60.11 °C from the initial temperature (peak temperature of 87.77 °C).

[0014] (b) An endothermic peak with an initial temperature of 145–155 °C and a peak temperature difference of 7–12 °C from the initial temperature; for example, crystal form I has an endothermic peak with an initial temperature of 151.39 °C and a peak temperature difference of 9.35 °C from the initial temperature (peak temperature of 160.74 °C).

[0015] (c) An endothermic peak with an initial temperature of 185–190 °C and a peak temperature difference of 2–8 °C from the initial temperature; for example, crystal form I has an endothermic peak with an initial temperature of 188.90 °C and a peak temperature difference of 4.19 °C from the initial temperature (peak temperature of 193.09 °C).

[0016] According to an exemplary embodiment of the present invention, the crystal form I has a DSC pattern as shown in FIG3.

[0017] According to an embodiment of the present invention, the crystal form II, using Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 7.8±0.20°, 11.2±0.20°, 12.5±0.20°, 16.9±0.20°, 20.1±0.20°, and 23.4±0.20° in 2θ angles; furthermore, characteristic peaks may also be observed at 5.6±0.20° and 13.3±0.20°. Characteristic peaks are present at 0.20°, 21.4±0.20°, 22.6±0.20°, 25.1±0.20° and / or 26.1±0.20°; furthermore, characteristic peaks are also present at 14.2±0.20°, 17.7±0.20°, 18.2±0.20°, 19.2±0.20°, 27.9±0.20° and / or 32.7±0.20°.

[0018] Alternatively, the crystal form II, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 5.6±0.20°, 7.8±0.20°, 11.2±0.20°, 12.5±0.20°, 16.9±0.20°, 20.1±0.20°, and 23.4±0.20° in 2θ angles; further, it may also exhibit characteristic peaks at 21.4±0.20°, 22.6±0.20°, 25.1±0.20°, and / or 26.1±0.20°; even further, it may also exhibit characteristic peaks at 14.2±0.20°, 17.7±0.20°, 18.2±0.20°, 19.2±0.20°, 27.9±0.20°, and / or 32.7±0.20°.

[0019] In one embodiment, the crystal form II is subjected to Cu-Kα radiation, and X-ray powder diffraction, expressed in 2θ angles, at 4.0±0.20°, 4.6±0.20°, 6.1±0.20°, 7.8±0.20°, 10.1±0.20°, 11.2±0.20°, 12.5±0.20°, 13.3±0.20°, 14.2±0.20°, 15.0±0.20°, 15.7±0.20°, 16.9±0.20°, and 17.7±0. Characteristic peaks are present at 0.20°, 18.2±0.20°, 19.2±0.20°, 20.1±0.20°, 21.4±0.20°, 22.0±0.20°, 22.6±0.20°, 23.4±0.20°, 25.1±0.20°, 26.1±0.20°, 27.9±0.20°, 30.2±0.20°, 30.8±0.20°, 31.3±0.20°, 31.8±0.20°, and 32.8±0.20°.

[0020] According to an embodiment of the present invention, the crystal form II has an XRPD pattern as shown in Figure 5, Figure 19 or Figure 22.

[0021] According to an embodiment of the present invention, the crystal form II loses no more than 3 wt% of weight at room temperature to about 170°C, for example, 2.4 wt%.

[0022] According to an embodiment of the present invention, the crystal form II is a hydrate, for example, a 0.5 to 0.8 hydrate.

[0023] According to an exemplary embodiment of the present invention, the crystal form II has a TGA pattern as shown in FIG6, FIG21 or FIG24.

[0024] According to an embodiment of the present invention, the crystal form II has one or two peaks as shown in (1)-(2):

[0025] (1) An endothermic peak with an initial temperature of 55-80℃ and a peak temperature difference of 18-33℃ from the initial temperature; for example, an endothermic peak with an initial temperature of 55-60℃ and a peak temperature difference of 18-26℃ from the initial temperature; or an endothermic peak with an initial temperature of 60-75℃ and a peak temperature difference of 18-30℃ from the initial temperature.

[0026] For example, crystal form II has an endothermic peak with an initial temperature of 58.97°C and a peak temperature difference of 22.17°C from the initial temperature (peak temperature is 81.14°C).

[0027] Alternatively, crystal form II has an endothermic peak with an initial temperature of 61.02°C and a peak temperature 25.57°C lower than the initial temperature (peak temperature 86.59°C).

[0028] Alternatively, crystal form II has an endothermic peak with an initial temperature of 69.33°C and a peak temperature 26.4°C lower than the initial temperature (peak temperature 95.73°C).

[0029] (2) An endothermic peak with an initial temperature of 188-196℃ and a peak temperature difference of 1-5℃ from the initial temperature;

[0030] For example, crystal form II has an endothermic peak with an initial temperature of 192.29°C and a peak temperature difference of 2.56°C from the initial temperature (peak temperature of 194.85°C).

[0031] Alternatively, crystal form II has an endothermic peak with an initial temperature of 193.02℃ and a peak temperature 1.91℃ higher than the initial temperature (peak temperature 194.93℃).

[0032] Alternatively, crystal form II has an endothermic peak with an initial temperature of 192.82°C and a peak temperature 1.8°C higher than the initial temperature (peak temperature 194.62°C).

[0033] According to an exemplary embodiment of the present invention, the crystal form II has a DSC pattern as shown in FIG7, FIG20 or FIG23.

[0034] According to an embodiment of the present invention, the crystal form III, using Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 9.8±0.20°, 14.7±0.20°, 16.7±0.20°, 19.6±0.20°, and 22.5±0.20° in 2θ angles; further, it may also have characteristic peaks at 11.2±0.20°, 12.4±0.20°, and / or 24.1±0.20°; even further, it may also have characteristic peaks at 12.9±0.20°, 21.4±0.20°, 23.1±0.20°, 23.4±0.20°, and / or 27.1±0.20°.

[0035] In one embodiment, the crystal form III is subjected to Cu-Kα radiation, and X-ray powder diffraction, expressed in 2θ angles, at 2.4±0.20°, 3.2±0.20°, 3.9±0.20°, 4.8±0.20°, 5.3±0.20°, 6.2±0.20°, 7.1±0.20°, 7.8±0.20°, 8.4±0.20°, 9.8±0.20°, 10.5±0.20°, 11.2±0.20°, 12.4±0.20°, 12.9±0.20°, 13.3±0.20°, 14.0±0.20°, 14.7±0.20°, 15.1±0.20°, and 15. Characteristic peaks are present at 8±0.20°, 16.1±0.20°, 16.7±0.20°, 17.2±0.20°, 17.8±0.20°, 18.4±0.20°, 19.6±0.20°, 20.7±0.20°, 21.4±0.20°, 22.5±0.20°, 23.1±0.20°, 23.4±0.20°, 24.1±0.20°, 24.5±0.20°, 24.8±0.20°, 25.6±0.20°, 26.0±0.20°, 26.4±0.20°, 27.1±0.20°, 27.8±0.20°, and 30.2±0.20°.

[0036] According to an embodiment of the present invention, the crystal form III has an XRPD pattern as shown in FIG9.

[0037] According to an embodiment of the present invention, the crystal form III loses no more than 2 wt% at room temperature to about 100°C, for example, 1.14 wt%; and loses 5 to 6.5 wt% at 100 to 140°C, for example, 5.7 wt%.

[0038] According to an exemplary embodiment of the present invention, the crystal form III has a TGA pattern as shown in FIG10.

[0039] According to an embodiment of the present invention, crystal form III has one, two, or three peaks as shown in (i)-(iii):

[0040] (i) An endothermic peak with an initial temperature of 45–52 °C and a peak temperature difference of 18–23 °C from the initial temperature; for example, crystal form III has an endothermic peak with an initial temperature of 48.24 °C and a peak temperature difference of 20.25 °C from the initial temperature (peak temperature of 68.49 °C).

[0041] (ii) An endothermic peak with an initial temperature of 115–125 °C and a peak temperature difference of 2–8 °C from the initial temperature; for example, crystal form III has an endothermic peak with an initial temperature of 119.57 °C and a peak temperature difference of 4.07 °C from the initial temperature (peak temperature of 123.64 °C).

[0042] (iii) An endothermic peak with an initial temperature of 185–195 °C and a peak temperature difference of 1–5 °C from the initial temperature; for example, crystal form III has an endothermic peak with an initial temperature of 189.46 °C and a peak temperature difference of 2.54 °C from the initial temperature (peak temperature of 192.00 °C).

[0043] According to an exemplary embodiment of the present invention, the crystal form III has a DSC pattern substantially as shown in FIG11.

[0044] According to an embodiment of the present invention, the crystal form IV, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 5.7±0.20°, 7.9±0.20°, 11.4±0.20°, 12.7±0.20°, and 17.1±0.20° in 2θ angles; further, it may also exhibit characteristic peaks at 20.4±0.20°, 22.8±0.20°, and / or 23.5±0.20°; even further, it may also exhibit characteristic peaks at 14.1±0.20°. Characteristic peaks are present at 15.5±0.20°, 17.6±0.20°, 18.0±0.20°, 19.3±0.20°, 21.5±0.20°, 23.9±0.20°, 24.8±0.20°, 25.4±0.20°, 26.3±0.20°, 27.4±0.20°, 28.4±0.20°, 30.8±0.20°, 31.6±0.20°, 33.3±0.20° and / or 36.6±0.20°.

[0045] In one embodiment, the crystal form IV is irradiated using Cu-Kα radiation, and X-ray powder diffraction is performed at angles of 2θ at 5.7±0.20°, 7.9±0.20°, 11.4±0.20°, 12.7±0.20°, 14.1±0.20°, 15.5±0.20°, 17.1±0.20°, 17.6±0.20°, 18.0±0.20°, 19.3±0.20°, and 20.4± Characteristic peaks are present at 0.20°, 21.5±0.20°, 22.8±0.20°, 23.5±0.20°, 23.9±0.20°, 24.8±0.20°, 25.4±0.20°, 26.3±0.20°, 27.4±0.20°, 28.4±0.20°, 30.8±0.20°, 31.6±0.20°, 33.3±0.20°, and 36.6±0.20°.

[0046] According to an embodiment of the present invention, the crystal form IV has an XRPD pattern as shown in FIG14.

[0047] According to an embodiment of the present invention, the crystal form IV has an endothermic peak with an initial temperature of 185-195°C and a peak temperature difference of 1-7°C from the initial temperature; for example, the crystal form IV has an endothermic peak with an initial temperature of 186.58°C and a peak temperature difference of 4.05°C from the initial temperature (peak temperature is 190.63°C).

[0048] According to an embodiment of the present invention, the crystal form IV also has an exothermic peak, for example, an onset temperature of not less than 200°C.

[0049] According to an exemplary embodiment of the present invention, the crystal form IV has a DSC pattern substantially as shown in FIG15.

[0050] According to an embodiment of the present invention, the crystal form V, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 4.6±0.20°, 10.9±0.20°, 18.8±0.20°, 19.7±0.20°, 20.6±0.20°, and 23.7±0.20° in 2θ angles; further, characteristic peaks may also be observed at 7.2±0.20° and 14.6±0.20°. Characteristic peaks are present at 20°, 18.1±0.20° and / or 20.1±0.20°; further, characteristic peaks are also present at 17.1±0.20°, 17.5±0.20°, 21.3±0.20°, 22.0±0.20°, 24.3±0.20°, 26.5±0.20°, 28.2±0.20° and / or 29.1±0.20°.

[0051] According to an embodiment of the present invention, the crystal form V is subjected to Cu-Kα radiation, and X-ray powder diffraction, expressed in 2θ angles, at 4.6±0.20°, 7.2±0.20°, 10.9±0.20°, 11.6±0.20°, 12.1±0.20°, 14.1±0.20°, 14.6±0.20°, 15.2±0.20°, 17.1±0.20°, 17.5±0.20°, 18.1±0.20°, 18.8±0.20°, 19.7±0.20°, 20.1±0.20°, 20.6±0.20°, and 21.3±0.20°. Characteristic peaks are present at 21.6±0.20°, 22.0±0.20°, 22.2±0.20°, 23.0±0.20°, 23.7±0.20°, 24.3±0.20°, 25.4±0.20°, 25.8±0.20°, 26.1±0.20°, 26.5±0.20°, 27.6±0.20°, 28.2±0.20°, 29.1±0.20°, 30.8±0.20°, 31.8±0.20°, 33.1±0.20°, 33.4±0.20°, 34.2±0.20°, and 34.5±0.20°.

[0052] According to an embodiment of the present invention, the crystal form V has an XRPD pattern as shown in FIG16.

[0053] According to an embodiment of the present invention, the crystal form V has one or two endothermic peaks as follows:

[0054] An endothermic peak with an initial temperature of 30–40°C and a peak temperature difference of 10–20°C from the initial temperature; for example, an endothermic peak with an initial temperature of 36.21°C and a peak temperature difference of 15.3°C from the initial temperature (peak temperature of 51.51°C).

[0055] An endothermic peak with an initial temperature of 106–116℃ and a peak temperature difference of 0.5–5℃ from the initial temperature; for example, an endothermic peak with an initial temperature of 111.09℃ and a peak temperature difference of 2.05℃ from the initial temperature (peak temperature of 113.14℃).

[0056] According to an exemplary embodiment of the present invention, the crystal form V has a DSC pattern as shown in FIG17.

[0057] According to an embodiment of the present invention, the crystal form V loses 3-12 wt% weight at 30-130°C, preferably 5-10 wt%.

[0058] According to an exemplary embodiment of the present invention, the crystal form V has a TGA pattern as shown in FIG18.

[0059] The present invention also provides an amorphous form of the compound of formula (I), the amorphous form having an XRPD pattern substantially as shown in FIG12.

[0060] This invention also provides a single crystal of the above-mentioned crystal form II, belonging to the orthorhombic crystal system, with space group P212121, and cell parameters [missing information]. α = β = γ = 90°.

[0061] The present invention provides a crystal form (e.g., crystal form II) of the compound of formula (I), which, using Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 7.8±0.20°, 11.2±0.20°, 12.5±0.20°, 16.9±0.20°, 20.1±0.20°, and 23.4±0.20° in 2θ angles; further, characteristic peaks can also be observed at 13.3±0.20° and 21.4°. Characteristic peaks are present at ±0.20°, 22.6±0.20°, 25.1±0.20° and / or 26.1±0.20°; furthermore, characteristic peaks are also present at 14.2±0.20°, 17.7±0.20°, 18.2±0.20°, 19.2±0.20°, 27.9±0.20° and / or 32.7±0.20°; preferably, characterization is performed using an ARL Equinox 100 X-ray powder diffractometer with a 2θ scan angle from 0° to 35°, a voltage of 40 kV, and a current of 0.9 mA;

[0062] Alternatively, using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 5.6±0.20°, 7.8±0.20°, 11.2±0.20°, 12.5±0.20°, 16.9±0.20°, 20.1±0.20°, and 23.4±0.20°; furthermore, characteristic peaks can also be observed at 13.3±0.20°, 21.4±0.20°, and 2... Characteristic peaks are present at 2.6±0.20°, 25.1±0.20° and / or 26.1±0.20°; further, characteristic peaks are also present at 14.2±0.20°, 17.7±0.20°, 18.2±0.20°, 19.2±0.20°, 27.9±0.20° and / or 32.7±0.20°; preferably, characterization is performed using a Bruker D8 Advance X-ray powder diffractometer, with parameters shown in Table 7-1.

[0063] The present invention also provides a crystalline hemihydrate of the compound of formula (I),

[0064] Preferably, the crystalline hemihydrate is crystal form II or a single crystal of crystal form II.

[0065] The present invention also provides a method for preparing crystal form II of the compound of formula (I), comprising using the compound of formula (I) or crystal form I as raw material, using solvent one as solvent or dispersant, and separating to obtain crystal form II;

[0066] The solvent one may be selected from one or more of methanol, ethanol, isopropanol, n-propanol, n-butanol, acetone, 4-methyl-2-pentanone, ethyl acetate, isopropyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, dimethyl sulfoxide, acetonitrile, toluene, n-heptane, methylcyclohexane, dichloromethane, methyl tert-butyl ether, anisole, isopropyl ether, dioxane, and water. For example, the solvent one may be selected from methanol, ethanol, or a mixture of dimethyl sulfoxide, acetone, and water.

[0067] According to an embodiment of the present invention, the preparation method of crystal form II can be selected from methods such as slow volatilization, room temperature suspension, high temperature suspension, temperature cycling suspension, rapid cooling, slow cooling, forward dropping dissolution, reverse dropping dissolution, vapor diffusion, etc.

[0068] In one embodiment, the preparation method of crystal form II includes: forming a suspension of crystal form I and ethanol, stirring at 45-55°C for 1-5 hours, then cooling to room temperature and stirring for at least 8 hours, separating, and obtaining crystal form II.

[0069] The present invention also provides a pharmaceutical composition comprising a polymorph, amorphous, crystalline hemihydrate and / or a single crystal of the compound of formula (I) above.

[0070] According to embodiments of the present invention, the pharmaceutical composition further comprises one, two or more pharmaceutically acceptable excipients.

[0071] According to embodiments of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0072] According to an embodiment of the present invention, the additional therapeutic agent is selected from smegglutinin.

[0073] The present invention also provides the use of polymorphs, single crystals, crystalline hemihydrates or pharmaceutical compositions of the compounds shown in formula (I) above in the preparation of medicaments for treating NLRP3-mediated conditions and / or diseases, such as in the preparation of NLRP3 inhibitor medicaments.

[0074] According to an embodiment of the present invention, the pharmaceutical composition comprises a polymorph, amorphous, crystalline hemihydrate or single crystal of the compound of formula (I), and smegglutinin.

[0075] According to embodiments of the present invention, the condition and / or disease is, for example, an autoinflammatory febrile syndrome such as cold pyridine-associated periodic syndrome (CAPS), sickle cell disease, systemic lupus erythematosus (SLE), chronic liver disease, non-alcoholic steatohepatitis (NASH), gout, gouty arthritis, pericarditis, type I and type II diabetes and related complications (e.g., nephropathy, retinopathy), neuroinflammatory-related disorders (e.g., multiple sclerosis, brain infection, acute injury, neurodegenerative diseases, Alzheimer's disease), atherosclerosis and cardiovascular risks (e.g., hypertension), obesity, hidradenitis suppurativa, wound healing and scar formation, and cancer (e.g., colorectal cancer, lung cancer, myeloproliferative neoplasms, leukemia, myelofibrosis).

[0076] According to embodiments of the present invention, the use of the polymorph, amorphous, crystalline hemihydrate, single crystal of crystal form II, or pharmaceutical composition of the compound represented by formula (I) in the preparation of a medicament for treating NLRP3-mediated obesity.

[0077] The present invention also provides a method for treating NLRP3-mediated symptoms and / or diseases, comprising administering to a patient a preventive or therapeutically effective amount of a polymorph, amorphous, crystalline hemihydrate, single crystal of crystal form II, and / or pharmaceutical composition of the compound represented by formula (I) above.

[0078] According to an embodiment of the present invention, the pharmaceutical composition comprises a polymorph, amorphous, crystalline hemihydrate or single crystal of the compound of formula (I), and smegglutinin.

[0079] According to embodiments of the present invention, the NLRP3-mediated conditions and / or diseases are selected from: autoinflammatory febrile syndromes such as cold pyridine-associated periodic syndrome (CAPS), sickle cell disease, systemic lupus erythematosus (SLE), chronic liver disease, non-alcoholic steatohepatitis (NASH), gout, gouty arthritis, pericarditis, type I and type II diabetes and related complications (e.g., nephropathy, retinopathy), neuroinflammatory-related disorders (e.g., multiple sclerosis, brain infection, acute injury, neurodegenerative diseases, Alzheimer's disease), atherosclerosis and cardiovascular risks (e.g., hypertension), obesity, hidradenitis suppurativa, wound healing and scar formation, and cancers (e.g., colorectal cancer, lung cancer, myeloproliferative neoplasms, leukemia, myelofibrosis).

[0080] In some embodiments, the present invention also provides a method for treating obesity, comprising administering to a patient a preventive or therapeutically effective amount of a polymorph, amorphous, crystalline hemihydrate, single crystal of crystal form II, or pharmaceutical composition of the compound represented by formula (I) above.

[0081] According to embodiments of the present invention, a pharmaceutical composition comprising administering to a patient a preventive or therapeutically effective amount of a compound of formula (I), the pharmaceutical composition comprising a polymorph, amorphous, crystalline hemihydrate or a single crystal of crystal form II of the compound represented by formula (I), and smegglutinin. Beneficial effects

[0082] The present invention provides a polymorph of the compound of formula (I) which has good hygroscopicity, solubility, physical stability and / or chemical stability, and is suitable for pharmaceutical preparation.

[0083] Terminology Definitions and Explanations

[0084] The term "pharmaceuticalally acceptable excipient" refers to an excipient that does not cause significant irritation to the organism and does not impair the biological activity and properties of the active compound.

[0085] The term "two or more" refers to two, three, four, five or more kinds.

[0086] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0087] The term “therapeutic effective amount” refers to the amount of an active compound or drug that researchers, veterinarians, physicians, or other clinicians are searching for in tissues, systems, animals, individuals, or humans to elicit a biological or medical response. It includes one or more of the following: (1) prevention of disease: e.g., prevention of disease, disorder, or condition in individuals susceptible to disease, disorder, or symptom but not yet experiencing or exhibiting the pathology or symptoms of the disease; (2) suppression of disease: e.g., suppression of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptoms of the disease, disorder, or symptom (i.e., prevention of further development of the pathology and / or symptoms); (3) relief of disease: e.g., relief of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptom of the disease, disorder, or symptom (i.e., reversal of the pathology and / or symptom).

[0088] The term "room temperature" refers to a temperature of 15 to 40°C, such as 20 to 35°C, or 25°C.

[0089] The term “about” represents an error of ±3°C, for example, ±2°C.

[0090] The compound shown in formula (I) is the compound of formula (I).

[0091] The term “characteristic peak at… and / or” means having a peak at one or more of the 2θ angles. Attached Figure Description

[0092] Figure 1 shows the XRPD pattern of crystal form I of compound (I);

[0093] Figure 2 shows the TGA spectrum of crystal form I of compound (I);

[0094] Figure 3 shows the DSC spectrum of crystal form I of compound (I);

[0095] Figure 4 shows the crystal form I of compound (I). 1 H NMR spectrum;

[0096] Figure 5 shows the XRPD pattern of crystal form II of compound (I).

[0097] Figure 6 shows the TGA spectrum of crystal form II of compound (I);

[0098] Figure 7 shows the DSC spectrum of crystal form II of compound (I);

[0099] Figure 8 shows the crystal form II of compound (I). 1 H NMR spectrum;

[0100] Figure 9 shows the XRPD pattern of crystal form III of compound (I);

[0101] Figure 10 shows the TGA spectrum of crystal form III of compound (I);

[0102] Figure 11 shows the DSC spectrum of crystal form III of compound (I);

[0103] Figure 12 shows the XRPD spectrum of the amorphous form of compound (I);

[0104] Figure 13 shows the DVS spectrum of crystal form II of compound (I);

[0105] Figure 14 shows the XRPD pattern of crystal form IV of compound (I);

[0106] Figure 15 shows the DSC spectrum of crystal form IV of compound (I);

[0107] Figure 16 shows the XRPD pattern of crystal form V of compound (I);

[0108] Figure 17 shows the DSC spectrum of crystal form V of compound (I);

[0109] Figure 18 shows the TGA spectrum of crystal form V of compound (I);

[0110] Figure 19 shows the XRPD pattern of the crystal form in Example 7 (Bruker D8 Advance test results);

[0111] Figure 20 shows the DSC spectrum of the crystal form in Example 7;

[0112] Figure 21 shows the TGA spectrum of the crystal form in Example 7;

[0113] Figure 22 shows the XRPD pattern of the crystal form in Example 7 (ARL Equinox 100 test results);

[0114] Figure 23 shows the DSC spectrum of the crystal form in Example 7 (test results of METTLER TOLEDO TGA2);

[0115] Figure 24 shows the TGA spectrum of the crystal form in Example 7 (METTLER TOLEDO DSC3 test results);

[0116] Figure 25 shows the PXRD pattern obtained from the single-crystal diffraction data of compound (I) crystal form II.

[0117] Figure 26 shows the effects of compound (I), smegglutinin, and their combination on mouse body weight. Detailed Implementation

[0118] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0119] Patent application PCT / CN2023 / 142422 (filed on December 27, 2023) discloses the compound shown in formula (I), and all contents involved in that patent application are incorporated herein by reference.

[0120] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0121] Nuclear magnetic resonance analysis (NMR) 1 H NMR)

[0122] Several milligrams of solid sample were dissolved in dimethyl sulfoxide-d6 or deuterated methanol solvent and analyzed by nuclear magnetic resonance on a Bruker AVANCE NEO 400 (Bruker, GER).

[0123] X-ray powder diffraction (XRPD) of crystalline forms I, II, III and amorphous forms

[0124] The obtained solid samples were analyzed using an ARL Equinox 100 X-ray powder diffractometer. The 2θ scanning angle ranged from 0° to 35°. The testing method was Cu target Kα radiation, voltage 40 kV, current 0.9 mA, and the sample disk was a zero-background sample disk.

[0125] Crystal form IV was determined using variable-temperature XRPD with a Panalytical Empyrean X-ray diffractometer. A suitable sample was evenly spread on a single-crystal silicon sample disk, and XRPD tests were performed using the following parameters:

[0126] Thermogravimetric analysis (TGA)

[0127] The thermogravimetric analyzer was a METTLER TOLEDO TGA2 (METTLER TOLEDO, US). 2-10 mg of sample was placed in a pre-equilibrated open aluminum sample pan and automatically weighed inside the TGA furnace. The sample was heated to the final temperature at a rate of 10 °C / min, with nitrogen purging at 50 mL / min at the sample location and 20 mL / min at the balance location.

[0128] Differential scanning calorimetry (DSC)

[0129] The differential scanning calorimeter was a METTLER TOLEDO DSC3 (METTLER TOLEDO, US). 2-10 mg of sample was accurately weighed and placed in a perforated DSC Tzero sample pan. The sample was heated to 80°C or 100°C at a rate of 2°C / min, held for 5 minutes, then cooled to 25°C at a rate of 10°C / min, held at 25°C for 10 minutes, and then heated to the final temperature at a rate of 2°C / min. Nitrogen purging was performed at a rate of 50 mL / min.

[0130] Dynamic moisture adsorption-desorption analysis (DVS)

[0131] Dynamic moisture adsorption-desorption analysis was performed using DVS Intrinsic (SMS, UK). The test adopted a gradient mode with humidity changes ranging from 0% to 95% to 0%. Within the range of 0% to 90%, the humidity change for each gradient was 10%. The gradient endpoint was determined using the dm / dt method, with a dm / dt value less than 0.002% and maintained for 10 minutes as the gradient endpoint.

[0132] High-performance liquid chromatography (HPLC)

[0133] The high-performance liquid chromatographs used were Waters Arc HPLC and Agilent 1260II, and the test conditions are shown in Tables 1 and 2.

[0134] Table 1. HPLC Test Conditions (Reversed Phase)

[0135] Table 2 HPLC Test Conditions (Normal Phase)

[0136] Example 1: Formula (I) Compound Crystal Form I

[0137] Preparation of (S)-3-((7-bromo-4-chlorophthalazine-1-yl)amino)propane-1,2-diol (IB) in step one

[0138] Compound IA (4 g, 0.014 mol) and (S)-3-aminopropane-1,2-diol (1.31 g, 0.014 mmol) were dissolved in ethanol (25 mL), and the mixture was heated to 80 °C and reacted for 4 hours. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by silica gel column chromatography using a dichloromethane / methanol eluent system to give the title compound IB (1.2 g, yield: 25%).

[0139] MS m / z(ESI): 332.1(M+1).

[0140] The second step is the preparation of (S)-3-((4-chloro-7-(prop-1-yn-1-yl)phthalazin-1-yl)amino)propane-1,2-diol (IC).

[0141] Compound IB (600 mg, 1.98 mmol), 1-(trimethylsilyl)propyne (178 mg, 1.59 mmol), cuprous iodide (151 mg, 0.79 mmol), cesium carbonate (646 mg, 1.98 mmol), and bis(triphenylphosphine)palladium dichloride (278 mg, 0.39 mmol) were dissolved in a mixed solvent of N,N-dimethylacetamide and water (22 mL, V / V = 10:1). The mixture was heated to 100 °C and reacted for 4 hours. After the reaction was completed, the reaction solution was added to a saturated ammonium chloride solution and extracted with ethyl acetate (10 mL × 3). The crude product was obtained by filtration and concentration. The residue was purified by silica gel column chromatography with dichloromethane / methanol as the eluent to give the title compound IC (70 mg, yield: 13.5%).

[0142] MS m / z(ESI):292.1(M+1).

[0143] Step 3: Preparation of (S)-3-((4-(4-chloro-2-hydroxyphenyl)-7-(prop-1-yn-1-yl)phthalazin-1-yl)amino)propane-1,2-diol (I)

[0144] Compound IC (50 mg, 0.17 mmol) was dissolved in a mixed solvent of dioxane and water (2.2 mL, V / V = 10:1), and (4-chloro-2-hydroxyphenyl)boronic acid (30 mg, 0.17 mmol), 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (12 mg, 0.02 mmol) and potassium carbonate (47 mg, 0.34 mmol) were added. The reaction mixture was stirred at 100 °C for 1 hour under nitrogen protection. After the reaction was completed, the reaction solution was filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography (Waters MS triggered Prep-LC with Acquity QDA detector, column: Welch 10μm C18 250×21.2mm; mobile phase 1: water (containing 0.1% NH3); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 15%-100%, flow rate: 25mL / min) to obtain compound I, namely compound (I) (4.4mg, yield: 6.7%).

[0145] MS m / z(ESI): 384.1(M+1) + .

[0146] 1 H NMR(400MHz,CD3OD)δ8.25(s,1H),7.72(dd,1H),7.55(d,1H),7.29(d,1H),7.01(d,1H),6.9 9(s,2H),4.02-3.95(m,1H),3.85-3.79(m,1H),3.75-3.69(m,1H),3.61(d,2H),2.10(s,3H).

[0147] 40 mg of the compound of formula (I) was weighed and added to 0.2 mL of methanol to form a suspension. The suspension was cyclically heated between 50°C and 5°C. Finally, the suspension was centrifuged at 5°C. The obtained substance was characterized by XRPD to be a crystalline form and named crystalline form I. The XRPD, TGA, DSC, and NMR spectra of crystalline form I of the compound of formula (I) are shown in Figures 1-4. The XRPD resolution results are shown in Table 3.

[0148] Table 3

[0149] TGA results showed that crystal form I experienced approximately 2.7% weight loss during heating to 160°C, and decomposition may occur above 240°C. DSC results showed that crystal form I exhibited a broad endothermic peak at an initial temperature of 27.66°C and a peak temperature of approximately 87.8°C, as well as endothermic peaks at an initial temperature of 151.39°C and a peak temperature of approximately 160.7°C, and at an initial temperature of 188.90°C and a peak temperature of approximately 193.1°C. 1 No obvious organic solvent signal peaks were observed in the 1H NMR results.

[0150] Example 2 Crystal form II of compound (I)

[0151] 40 mg of crystal form I was weighed as raw material and 0.4 mL of ethanol was added to form a suspension. After stirring at 50 °C for 2 hours, the suspension was cooled to room temperature and stirred for one day. After centrifugation, a solid was obtained. XRPD characterization (see Figure 5) showed that it was crystal form II of compound (I).

[0152] The XRPD analysis results for crystal form II are shown in Table 4. TGA results show that crystal form II experiences approximately 2.4% weight loss during heating to 170℃ and may decompose after 240℃ (see Figure 6). DSC results show endothermic peaks for crystal form II at an initial temperature of 58.97℃ and a peak temperature of approximately 81.1℃, and at an initial temperature of 192.29℃ and a peak temperature of approximately 194.9℃ (see Figure 7). 1 1H NMR results and reference spectrum (crystal type I) 1 The HNMR spectra were basically consistent; a solvent residue peak of ethanol was observed at 1.05 ppm, and the molar ratio of ethanol was calculated to be 0.07 eq (see Figure 8). As shown in Figure 13, crystal form II showed a 2.62% increase in weight due to moisture absorption at 80% humidity and a 3.33% increase at 95% humidity; and the crystal form remained unchanged before and after the DVS test.

[0153] Table 4

[0154] Example 3 Crystal form III of compound (I)

[0155] The crystal form III of compound (I) was obtained by slow evaporation in methanol: 5 mg of crystal form I was weighed as raw material, dissolved in 0.6 mL of methanol, and slowly evaporated at room temperature to obtain a solid. A wet sample was taken for testing, and the crystal form was characterized by XRPD (see Figure 9). It was recorded as the crystal form III of compound (I).

[0156] The XRPD analysis results for crystal form III are shown in Table 5. TGA results show that crystal form III experiences approximately 1.14% weight loss upon heating to 100℃, and approximately 5.7% weight loss during heating from 100 to 140℃. Decomposition may occur after 250℃ (see Figure 10). DSC results show endothermic peaks for crystal form III at the following temperatures: onset temperature 48.24℃, peak temperature approximately 68.5℃; onset temperature 119.57℃, peak temperature approximately 123.6℃; and onset temperature 189.46℃, peak temperature approximately 192.0℃ (see Figure 11).

[0157] Table 5

[0158] Example 4: Amorphous form of compound (I)

[0159] 20 mg of crystal form I was dissolved in 8 ml of acetonitrile, and then added to 16 ml of n-heptane. The resulting clear solution was allowed to evaporate slowly. The solid was characterized by XRPD (Figure 12), which showed that it was an amorphous form.

[0160] Example 5: Compound (I) Crystal Form IV

[0161] 20 mg of crystal form II was heated to 130 °C at a rate of 2 °C / min under nitrogen purging, and XRPD was tested to obtain crystal form IV. The XRPD pattern of crystal form IV is shown in Figure 14, and the analysis results are shown in Table 6.

[0162] Table 6

[0163] Crystal form II was heated to 130°C at a rate of 2°C / min in DSC, cooled to room temperature, and then heated to 300°C at a rate of 10°C / min to obtain the DSC spectrum of crystal form IV (see Figure 15). The spectrum shows that crystal form IV has an endothermic peak at the initial temperature of 186.58°C and the peak temperature of 190.63°C.

[0164] The partial test information for the crystal forms in Examples 6 and 7 is as follows:

[0165] X-ray powder diffraction (XRPD)

[0166] XRPD diffraction patterns were acquired using an X-ray diffractometer. Samples were prepared on a zero-background silicon wafer by gently pressing the sample onto a flat surface. The parameters of the XRPD diffraction are listed in Table 7-1.

[0167] Table 7-1 Parameters for XRPD testing using Bruker D8 Advance

[0168] Thermogravimetric analysis (TGA)

[0169] Thermogravimetric analysis (TGA) was performed using a TA instrument. Approximately 1–3 mg of sample was loaded into a pre-weighed aluminum pan and heated according to the parameters in Table 7-2. The data were analyzed using TRIOS software.

[0170] Table 7-2 Thermogravimetric Analysis Test Parameters

[0171] Differential scanning calorimetry (DSC)

[0172] Differential scanning calorimetry (DSC) analysis was performed using a TA instrument. Approximately 1–3 mg of sample was placed in an aluminum dish with a pinhole and heated according to the parameters in Table 7-3. Data were analyzed using TRIOS software.

[0173] Table 7-3 DSC Test Parameters

[0174] Hydrogen nuclear magnetic resonance (HNMR) 1 H-NMR)

[0175] Data was acquired using a Bruker 400MHz instrument. 1 ¹H-NMR spectra. Unless otherwise specified, samples were prepared in CD3OD (deuterated methanol) solvent and tested according to the parameters in Table 7-4. Data were analyzed using MestReNova software.

[0176] Table 7-4 1 H-NMR analysis parameters

[0177] High-performance liquid chromatography (HPLC)

[0178] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent HPLC 1260 series instrument. The methods are listed in Table 7-5.

[0179] Table 7-5 High Performance Liquid Chromatography Methods

[0180] KF

[0181] KF analysis was performed using a Mettler V30S KF titrator series instrument. The methods are listed in Table 7-6.

[0182] Table 7-6 KF Test Method

[0183] Example 6: Formula (I) Compound Crystal Form V

[0184] Preparation method of crystalline form V of compound (I): 100 mg of compound (I) was added to 2 ml of a mixed solvent of dimethyl sulfoxide / butanone / hexaethylphosphoric triamine (1 / 9 / 10, v / v / v), stirred overnight at 25 °C, centrifuged, and the solid was dried in a vacuum oven at 50 °C to obtain the product. XRPD characterization confirmed its crystalline form, and it was named crystalline form V. The XRPD, DSC, and TGA spectra of crystalline form V are shown in Figures 16-18, and the XRPD resolution results are shown in Table 8. The DSC results show that crystalline form V has an endothermic peak with an initial temperature of 36.21 °C and a peak temperature of 51.51 °C, as well as an endothermic peak with an initial temperature of 111.09 °C and a peak temperature of 113.14 °C. The TGA results show that crystalline form V loses approximately 8.4 wt% of weight between 30 and 130 °C.

[0185] Table 8

[0186] Example 7: Crystal form of compound (I)

[0187] 100 mg of compound (I) was added to 1.8 mL of a mixed solvent of dimethyl sulfoxide / acetone / water (2.7 / 5.3 / 10, v / v / v), stirred overnight at 25 °C, centrifuged, and the solid was dried in a vacuum oven at 50 °C to obtain the product. XRPD characterization showed it to be a crystalline form (Figure 19) and a hemihydrate. The XRPD analysis results are shown in Table 9.

[0188] Table 9

[0189] Based on the following test results, it can be seen that the crystal form obtained in this embodiment is crystal form II.

[0190] DSC results showed that this crystal form exhibited two endothermic peaks: one with an initial temperature of 61.02℃ and a peak temperature of 86.59℃, and the other with an initial temperature of 193.02℃ and a peak temperature of 194.93℃ (Figure 20). TGA results showed that this crystal form lost approximately 2.1 wt% of weight between 50 and 120℃ and approximately 0.2 wt% of weight between 180 and 215℃ (Figure 21). KF analysis indicated a water content of 2.7%. HPLC testing showed a sample purity of 99.62%. 1 H-NMR showed that the sample contained 0.15% acetone and 0.07% dimethyl sulfoxide.

[0191] The crystal form of this embodiment was analyzed using an ARL Equinox 100 X-ray powder diffractometer (2θ scanning angle from 0° to 35°; test method: Cu target Kα rays, voltage 40 kV, current 0.9 mA, sample disk with zero background). The XRPD pattern is shown in Figure 22, and the analysis results are shown in Table 10. The results show that the XRPD pattern of the crystal form in this embodiment is consistent with that of crystal form II in Example 2.

[0192] Table 10

[0193] The following instruments and methods were used to further analyze the crystal form of this embodiment using DSC and TGA. The DSC results showed that the crystal form had an endothermic peak with an initial temperature of 69.33℃ and a peak temperature of 95.73℃, as well as an endothermic peak with an initial temperature of 192.82℃ and a peak temperature of 194.62℃ (Figure 23). The TGA results showed that the crystal form lost approximately 2.4 wt% of weight between 50 and 120℃ (Figure 24). The results indicate that the DSC and TGA spectra of the crystal form in this embodiment are essentially consistent with those of crystal form II in Example 2.

[0194] Thermogravimetric analysis (TGA)

[0195] The thermogravimetric analyzer was a METTLER TOLEDO TGA2 (METTLER TOLEDO, US). 2-10 mg of sample was placed in a pre-equilibrated open aluminum sample pan and automatically weighed inside the TGA furnace. The sample was heated to the final temperature at a rate of 10 °C / min, with nitrogen purging at 50 mL / min at the sample location and 20 mL / min at the balance location.

[0196] Differential scanning calorimetry (DSC)

[0197] The differential scanning calorimeter was a METTLER TOLEDO DSC3 (METTLER TOLEDO, US). 2-10 mg of sample was accurately weighed and placed in a perforated DSC Tzero sample pan. The sample was heated to 80°C or 100°C at a rate of 2°C / min, held for 5 minutes, then cooled to 25°C at a rate of 10°C / min, held at 25°C for 10 minutes, and then heated to the final temperature at a rate of 2°C / min. Nitrogen purging was performed at a rate of 50 mL / min.

[0198] Example 8 Single crystal of crystal form II

[0199] Add 5 mg of compound (I) to 0.6 ml of methanol (MeOH), dissolve at room temperature, filter, and slowly evaporate the filtrate at room temperature to obtain crystals.

[0200] The following characterization confirmed that the obtained crystal is a single crystal of crystal type II.

[0201] Using the following single-crystal X-ray diffractometer, the single-crystal diffraction data was used to simulate the PXRD pattern shown in Figure 25.

[0202] Bruker D8 VENTURE dual-microspot single-crystal X-ray diffractometer, ambient temperature 223K, enhanced Cu light source, wavelength

[0203] The crystal analysis data are shown in Table 11.

[0204] Table 11

[0205] Stability Study of Test Example 1

[0206] Stability studies were conducted on crystal form II under high temperature (60℃), high humidity (25℃ / 92.5% RH), and light irradiation (25℃ / 4500 Lux) conditions. Samples were taken on day 7 for XRPD characterization and HPLC testing. The results are shown in Table 12.

[0207] Table 12

[0208] The stability of crystal form IV was tested under the same conditions described above. Crystal form IV exhibited similar, identical, or better stability than crystal form II.

[0209] Test Example 2: Biological Media and Water Solubility Test

[0210] The preparation process of the biological medium is shown in Table 13. Crystal form II samples were added to the biological medium and water and stirred at 37°C for 24 h. Samples were taken at 0.5 h, 2 h, and 24 h. The sampled solutions were filtered through a 0.22 μm aqueous filter membrane. Samples with higher concentrations were appropriately diluted with diluent. The peak area of ​​the solution was measured by HPLC. Finally, the concentration of the compound in the solution was calculated based on the peak area, the HPLC standard curve of the raw material, and the dilution factor. In addition, the pH value of the supernatant after 24 h was tested. The results are shown in Table 14. Crystal form II has high solubility in FaSSGF (pH = 1.6), and the remaining solid crystal form remained unchanged in all systems.

[0211] Table 13 Preparation process of biological media

[0212] Table 14

[0213] Test Example 3: Pharmacological Experiment of a High-Fat Diet-Induced DIO Obese Mouse Model

[0214] 3.1 Experimental Objective

[0215] The efficacy of the compound on a high-fat diet-induced DIO mouse model was tested.

[0216] 3.2 Test Materials

[0217] 3.2.1 Reagents

[0218] Hematoxylin staining solution: Zhongshan Jinqiao Biotechnology Co., Ltd., Product No.: ZLI-9610. Water-soluble eosin staining solution: Zhongshan Jinqiao Biotechnology Co., Ltd., Product No.: ZLI-9613. Sirius red staining solution: Beijing Solarbio Technology Co., Ltd., Product No.: G1427.

[0219] 3.2.2 Equipment

[0220] Electronic balance: Sartorius, Model: Quintix 124-1CN

[0221] Electronic scale: Shanghai Yueping Scientific Instruments Co., Ltd., Model: YP10001

[0222] Ultrasonic cleaner: Kunshan Ultrasonic Instrument Co., Ltd., Model: KQ3200E

[0223] Oscillator: Qilinbell, Model: 01-00X-1012

[0224] High-speed homogenizer: IKA, Model: T10 basic

[0225] Anesthesia machine: MIP, Model: 50318

[0226] Refrigerated centrifuge: Thermo Fisher, part number: Sorvall Legend Micro 17r

[0227] Multifunctional microplate reader: TECAN, catalog number: SPARK 10M

[0228] Microscope: OLYMPUS, Product No.: DP720

[0229] Tissue dehydrator: Leica, Part No.: TP1020

[0230] Paraffin embedding machine: Leica, part number: EG1150H

[0231] Freezing station: Leica, item number: EG1150C

[0232] Slicer: Leica, Part No.: RM2235

[0233] 3.3 Experimental Methods

[0234] 3.3.1 Experimental Grouping

[0235] Table 15 Animal grouping and administration regimens

[0236] 3.3.2 Model Establishment and Drug Administration

[0237] One week after arrival and acclimatization, the animals were randomly divided into two groups (7+28): 7 mice were fed a normal diet, and 28 mice were fed a high-fat diet (HFD). After 22-23 weeks of rearing, the animals were fasted for 6 hours, and blood glucose levels were measured using a glucometer. Blood was collected from the orbital sinus of all animals, and serum was collected by centrifugation to measure serum total cholesterol (TC) and triglyceride (TG) levels. Based on body weight, serum TC, and TG levels, the animals were randomly divided into four groups: one group continued to be fed a high-fat diet (HFD), and the other group of 7 mice received a normal diet. Administered medications according to Table 4 for 30 days.

[0238] 3.3.3 Reagent Preparation

[0239] 1) Semaglutide stock solution (0.5 mg / ml): Weigh 1.0 mg of semaglutide, add 2 ml of PBS, and slowly pipette until dissolved to prepare a stock solution with a concentration of 0.5 mg / ml.

[0240] Semaglutide low dose (0.0125 mg / ml): Pipette 25 uL of the stock solution and add 975 uL of PBS to obtain a solution with a concentration of 0.0125 mg / ml. Prepare fresh before use.

[0241] 2) Preparation of 0.5% sodium carboxymethyl cellulose (CMC-Na) solution (for dissolving the test sample GS8-164): Weigh 0.5g of CMC-Na powder, slowly add 100mL of ultrapure water, stir magnetically and heat to dissolve and mix well. Filter through a 0.22mm filter membrane and store at 2-8℃ until use.

[0242] Compound (I) (2 mg / mL, content: 96.7%): Weigh 84.0 mg of compound (I), add 40.614 mL of 0.5% CMC-Na, add a magnetic stir bar and stir thoroughly on a magnetic stirrer, vortex, and sonicate for 20 minutes to make the drug preparation in a suspension state. Prepare once every two days and store at 2-8℃ for later use.

[0243] 3.4 Detection Indicators and Results

[0244] The drug was administered continuously for one month, and the patient's weight was measured every three days during the administration period. The experimental results are shown in Figure 26.

[0245] At a dose of 20 mpk of compound (I), the weight loss was 5% in 30 days compared to the model group. The weight loss effect of compound (I) (20 mpk) in combination with semaglutide (0.025 mpk) (33%) was better than that of semaglutide (0.025 mpk) alone (23%).

[0246] Conclusion: Compound (I) showed good weight loss effect when used alone in the DIO obesity model, and its weight loss effect was better than that of Semaglutide alone when used in combination with it, demonstrating excellent potential for combination therapy.

[0247] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A polymorph of the compound of formula (I), characterized in that, The polymorphs include crystal forms I, II, III, IV and V of the compound of formula (I); 2. The polymorph according to claim 1, characterized in that, The crystal form I, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 17.2±0.20°, 22.0±0.20°, 22.6±0.20°, 24.9±0.20°, 25.7±0.20°, and 27.4±0.20° in 2θ angles; further, it also exhibits characteristic peaks at 8.6±0.20°, 10.9±0.20°, 14.4±0.20°, 20.6±0.20°, and / or 29.7±0.20°; even further, it also exhibits characteristic peaks at 9.5±0.20°, 18.6±0.20°, 20.0±0.20°, 23.8±0.20°, 31.7±0.20°, and / or 32.7±0.20°. Preferably, the crystal form I has an XRPD pattern as shown in Figure 1; Preferably, the crystal form I loses no more than 4 wt% of weight at room temperature to about 160°C; Preferably, crystal form I is a hydrate; Preferably, the crystal form I has a TGA pattern as shown in Figure 2; Preferably, the crystal form I has one, two, or three peaks as shown in (a)-(c): (a) An endothermic peak with an initial temperature of 25–30°C and a peak temperature difference of 55–65°C from the initial temperature; (b) An endothermic peak with an initial temperature of 145–155 °C and a peak temperature difference of 7–12 °C from the initial temperature; (c) An endothermic peak with an initial temperature of 185–190 °C and a peak temperature difference of 2–8 °C from the initial temperature; Preferably, the crystal form I has a DSC pattern as shown in Figure 3.

3. The polymorph according to claim 1, characterized in that, The crystal form II, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 7.8±0.20°, 11.2±0.20°, 12.5±0.20°, 16.9±0.20°, 20.1±0.20°, and 23.4±0.20° in 2θ angles; further, it also exhibits characteristic peaks at 5.6±0.20°, 13.3±0.20°, 21.4±0.20°, 22.6±0.20°, 25.1±0.20°, and / or 26.1±0.20°; even further, it also exhibits characteristic peaks at 14.2±0.20°, 17.7±0.20°, 18.2±0.20°, 19.2±0.20°, 27.9±0.20°, and / or 32.7±0.20°. For example, the crystal form II has an XRPD pattern that is basically as shown in Figure 5, Figure 19 or Figure 22; For example, the crystal form II loses no more than 3 wt% of weight at room temperature to about 170°C; Preferably, the crystal form II is a hydrate, for example, a 0.5 hydrate; For example, the crystal form II has a TGA pattern as shown in Figure 6, Figure 21 or Figure 24; For example, crystal form II has one or two peaks as shown in (1)-(2): (1) An endothermic peak with an initial temperature of 55-80℃ and a peak temperature difference of 18-33℃ from the initial temperature; For example, an endothermic peak with an initial temperature of 55–60°C and a peak temperature difference of 18–26°C from the initial temperature; or an endothermic peak with an initial temperature of 60–75°C and a peak temperature difference of 18–30°C from the initial temperature. (2) An endothermic peak with an initial temperature of 188-196℃ and a peak temperature difference of 1-5℃ from the initial temperature; For example, the crystal form II has a DSC pattern as shown in Figure 7, Figure 20 or Figure 23.

4. The polymorph according to claim 1, characterized in that, The crystal form III, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 9.8±0.20°, 14.7±0.20°, 16.7±0.20°, 19.6±0.20°, and 22.5±0.20° in 2θ angles; further, it also exhibits characteristic peaks at 11.2±0.20°, 12.4±0.20°, and / or 24.1±0.20°; even further, it also exhibits characteristic peaks at 12.9±0.20°, 21.4±0.20°, 23.1±0.20°, 23.4±0.20°, and / or 27.1±0.20°. Preferably, the crystal form III has an XRPD pattern as shown in Figure 9; Preferably, the crystal form III loses no more than 2 wt% at room temperature to about 100°C, and loses 5 to 6.5 wt% at 100 to 140°C; Preferably, the crystal form III has a TGA pattern as shown in Figure 10; Preferably, crystal form III has one, two, or three peaks as shown in (i)-(iii): (i) an endothermic peak with an initial temperature of 45–52 °C and a peak temperature difference of 18–23 °C from the initial temperature; (ii) an endothermic peak with an initial temperature of 115–125°C and a peak temperature difference of 2–8°C from the initial temperature; (iii) An endothermic peak with an initial temperature of 185–195°C and a peak temperature difference of 1–5°C from the initial temperature; Preferably, the crystal form III has a DSC pattern as shown in Figure 11.

5. The polymorph according to claim 1, characterized in that, The crystal form IV, when irradiated with Cu-Kα, exhibits characteristic peaks in X-ray powder diffraction at 5.7±0.20°, 7.9±0.20°, 11.4±0.20°, 12.7±0.20°, and 17.1±0.20° in 2θ angles; further, it also exhibits characteristic peaks at 20.4±0.20°, 22.8±0.20°, and / or 23.5±0.20°; even further, it also exhibits characteristic peaks at 14.1±0.20° and 15.5±0.20°. Characteristic peaks are present at 20°, 17.6±0.20°, 18.0±0.20°, 19.3±0.20°, 21.5±0.20°, 23.9±0.20°, 24.8±0.20°, 25.4±0.20°, 26.3±0.20°, 27.4±0.20°, 28.4±0.20°, 30.8±0.20°, 31.6±0.20°, 33.3±0.20° and / or 36.6±0.20°. Preferably, the crystal form IV has an XRPD pattern as shown in Figure 14; Preferably, the crystal form IV has an endothermic peak with an initial temperature of 185–195°C and a peak temperature that differs from the initial temperature by 1–7°C. Preferably, the crystal form IV also has an exothermic peak; Preferably, the crystal form IV has a DSC pattern as shown in Figure 15.

6. The polymorph according to claim 1, characterized in that, The crystal form V, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 4.6±0.20°, 10.9±0.20°, 18.8±0.20°, 19.7±0.20°, 20.6±0.20°, and 23.7±0.20°, expressed in 2θ angles; furthermore, characteristic peaks can also be observed at 7.2±0.20°, 14.6±0.20°, and 1... Characteristic peaks are present at 8.1±0.20° and / or 20.1±0.20°; furthermore, characteristic peaks are also present at 17.1±0.20°, 17.5±0.20°, 21.3±0.20°, 22.0±0.20°, 24.3±0.20°, 26.5±0.20°, 28.2±0.20° and / or 29.1±0.20°. Preferably, the crystal form V has an XRPD pattern as shown in Figure 16; Preferably, the crystal form V has one or two endothermic peaks as follows: An endothermic peak with an initial temperature of 30–40℃ and a peak temperature difference of 10–20℃ from the initial temperature; An endothermic peak with an initial temperature of 106–116℃ and a peak temperature difference of 0.5–5℃ from the initial temperature; Preferably, the crystal form V has a DSC pattern as shown in Figure 17; Preferably, the crystal form V loses 3-12 wt% of weight at 30-130°C; Preferably, the crystal form V has a TGA pattern as shown in Figure 18.

7. The amorphous form of the compound of formula (I), characterized in that, The amorphous material has a basic XRPD pattern as shown in Figure 12; 8. A single crystal of crystal form II, characterized in that, The single crystal belongs to an orthorhombic crystal system, with space group P212121 and cell parameters [missing information]. α = β = γ = 90°; The crystal form II is the crystal form II in claim 3.

9. The crystalline hemihydrate of compound (I), Preferably, the crystalline hemihydrate is crystal form II as described in claim 3 or the single crystal as described in claim 8.

10. A method for preparing crystal form II of the compound of formula (I) according to claim 3, characterized in that, The preparation method includes using a compound of formula (I) or crystal form I as raw material, using solvent one as solvent or dispersant, and separating to obtain crystal form II; The solvent is selected from one or more of methanol, ethanol, isopropanol, n-propanol, n-butanol, acetone, 4-methyl-2-pentanone, ethyl acetate, isopropyl acetate, tetrahydrofuran, 2-methyltetrahydrofuran, dimethyl sulfoxide, acetonitrile, toluene, n-heptane, methylcyclohexane, dichloromethane, methyl tert-butyl ether, anisole, isopropyl ether, dioxane, and water.

11. A pharmaceutical composition, characterized in that, The polymorph of the compound of formula (I) according to any one of claims 1-6, the amorphous form according to claim 7, the single crystal according to claim 8, or the crystalline hemihydrate according to claim 9; Preferably, the pharmaceutical composition further comprises one, two or more pharmaceutically acceptable excipients; Preferably, the pharmaceutical composition further comprises one or more additional therapeutic agents, and the additional therapeutic agent is preferably smegglutinin.

12. Use of the polymorph of the compound of formula (I) according to any one of claims 1-6, the amorphous form according to claim 7, the single crystal according to claim 8, the crystalline hemihydrate according to claim 9, or the pharmaceutical composition according to claim 11 in the preparation of a medicament for treating NLRP3-mediated conditions and / or diseases, such as in the preparation of an NLRP3 inhibitor medicament; Preferably, the pharmaceutical composition comprises a polymorph of the compound of formula (I), the amorphous form of claim 7, the single crystal of claim 8, or the crystalline hemihydrate of claim 9, and smegglutinin.

13. The use according to claim 12, characterized in that, The conditions and / or diseases mentioned are autoinflammatory febrile syndromes such as cold pyrrolizum-associated periodic syndrome (CAPS), sickle cell disease, systemic lupus erythematosus (SLE), chronic liver disease, non-alcoholic steatohepatitis (NASH), gout, gouty arthritis, pericarditis, type I and type II diabetes and related complications (e.g., nephropathy, retinopathy), neuroinflammatory-related disorders (e.g., multiple sclerosis, brain infection, acute injury, neurodegenerative diseases, Alzheimer's disease), atherosclerosis and cardiovascular risks (e.g., hypertension), obesity, hidradenitis suppurativa, wound healing and scar formation, and cancers (e.g., colorectal cancer, lung cancer, myeloproliferative neoplasms, leukemia, myelofibrosis); for example, obesity.

14. A method for treating NLRP3-mediated symptoms and / or diseases, characterized in that, The method includes administering to a patient a preventive or therapeutically effective amount of a polymorph of the compound of formula (I) of any one of claims 1-6, an amorphous form of claim 7, a single crystal of claim 8, a crystalline hemihydrate of claim 9, and / or a pharmaceutical composition of claim 11.

15. The method according to claim 14, characterized in that, The conditions and / or diseases mentioned are autoinflammatory febrile syndromes such as cold pyridine-associated periodic syndrome (CAPS), sickle cell disease, systemic lupus erythematosus (SLE), chronic liver disease, non-alcoholic steatohepatitis (NASH), gout, gouty arthritis, pericarditis, type I and type II diabetes and related complications (e.g., nephropathy, retinopathy), neuroinflammatory-related disorders (e.g., multiple sclerosis, brain infection, acute injury, neurodegenerative diseases, Alzheimer's disease), atherosclerosis and cardiovascular risks (e.g., hypertension), obesity, hidradenitis suppurativa, wound healing and scar formation, and cancers (e.g., colorectal cancer, lung cancer, myeloproliferative neoplasms, leukemia, myelofibrosis).

16. A method for treating obesity, characterized in that, The pharmaceutical composition includes polymorphs of the compound of formula (I) according to any one of claims 1-6, the amorphous form according to claim 7, the single crystal according to claim 8, the crystalline hemihydrate according to claim 9, and / or the pharmaceutical composition according to claim 11, which are given to patients in a preventive or therapeutically effective amount. Preferably, the pharmaceutical composition comprises a polymorph of the compound of formula (I), the amorphous form of claim 7, the single crystal of claim 8, or the crystalline hemihydrate of claim 9, and smegglutinin.

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