NLRP3 inhibitor compound, preparation method therefor, and use thereof

Abstract

An NLRP3 inhibitor compound, a preparation method therefor, and use thereof. The compound has a structure represented by formula (A). In the formula, X represents an acid, and n is a number of 0.1-4. The compound has good bioactivity and solubility, showing promise as a drug.

Description

NLRP3 inhibitor compound and preparation method and application thereof

[0001] This application claims the priority of the prior application filed with the China National Intellectual Property Office on June 28, 2024, with the patent application number 2024108646951, and the invention name of "NLRP3 inhibitor compound and preparation method and application thereof"; the entire contents of the prior application are incorporated herein by reference. TECHNICAL FIELD

[0002] The present application belongs to the field of compounds, and particularly relates to a NLRP3 inhibitor compound and a preparation method and application thereof. BACKGROUND

[0003] NOD-like receptor protein 3 (NLRP3) is an important member of the NOD-like receptor family. NLRP3 contains three parts of pyrin domain (PYD), nucleotide-binding domain (NBD) and leucine-rich repeat (LRR). When the cell receives the stimulation of aseptic inflammatory danger signals, NLRP3 interacts with the adapter protein apoptosis-associated speck-like protein (ASC) and pro-caspase-1 to form the NLRP3 inflammasome complex. The activation of NLRP3 inflammasome leads to the release of IL-1β and IL-18.

[0004] There are few NLRP3 inflammasome inhibitors under research at present. It is necessary to develop NLRP3 inflammasome inhibitors with better pharmaceutical properties such as physicochemical properties, bioavailability, biological activity, etc., for clinical needs. SUMMARY

[0005] In order to improve the above technical problems, the present application provides a compound having the structure as shown in formula A:

[0006] wherein X represents an acid, and n is a number of 0.1-4.

[0007] According to an embodiment of the present application, the acid is an inorganic acid or an organic acid, for example, the inorganic acid is selected from one or two or more of hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, etc., and for example, the organic acid is selected from one or two or more of methanesulfonic acid, lactic acid, maleic acid, malic acid (for example, L-malic acid), tartaric acid (for example, L-tartaric acid), p-toluenesulfonic acid, fumaric acid, ethanesulfonic acid, malonic acid, and oxalic acid, etc.

[0008] According to embodiments of the application, n is a number from 0.4 to 3, for example 0.4, 0.5, 0.53, 0.55, 0.6, 0.7, 0.8, 0.9, 1.0, 1.01, 1.02, 1.03, 1.04, 1.05, 1.06, 1.07, 1.08, 1.09, 1.1, 1.11, 1.12, 1.13, 1.14, 1.15, 1.2, 1.21, 1.25, 1.3, 1.35, 1.4, 1.41, 1.42, 1.43, 1.44, 1.45, 1.5, 1.53, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8 or 2.9.

[0009] In certain embodiments, when X represents hydrochloric acid, n is a number from 0.7 to 1.3.

[0010] In certain embodiments, when X represents sulfuric acid, n is a number from 0.7 to 1.3.

[0011] In certain embodiments, when X represents phosphoric acid, n is a number from 0.5 to 2.5.

[0012] In certain embodiments, when X represents methanesulfonic acid, n is a number from 0.7 to 1.3.

[0013] In certain embodiments, when X represents hydrobromic acid, n is a number from 0.5 to 2.5.

[0014] In certain embodiments, when X represents maleic acid, n is a number from 0.9 to 1.3.

[0015] In certain embodiments, when X represents fumaric acid, n is a number from 0.4 to 1.3.

[0016] In certain embodiments, when X represents L-malic acid, n is a number from 0.5 to 1.3.

[0017] In certain embodiments, when X represents L-tartaric acid, n is a number from 0.5 to 1.3.

[0018] In certain embodiments, when X represents malonic acid, n is a number from 0.5 to 1.3.

[0019] In certain embodiments, when X represents p-toluene sulfonic acid, n is a number from 0.5 to 1.3.

[0020] According to embodiments of the application, the compound is a compound of formula A-I, a compound of formula A-II, a compound of formula A-III or a compound of formula A-IV;

[0021] The compound of formula A-I has the following structure:

[0022] n is selected from the group consisting of numbers from 0.6 to 1.8, preferably from 0.9 to 1.3, and exemplarily from 0.9 to 1 (e.g. 0.9);

[0023] The compound of formula A-II has the following structure:

[0024] n is selected from the group consisting of numbers from 0.6 to 1.8, preferably from 0.9 to 1.3, and exemplarily from 0.9 to 1.1 (e.g. 1);

[0025] The compound of formula A-III has the following structure:

[0026] n is selected from the group consisting of numbers from 0.5 to 1.5, preferably from 0.9 to 1.3, and exemplarily 1;

[0027] The compound of formula A-IV has the following structure:

[0028] n is selected from the group consisting of numbers from 0.5 to 1.5, preferably from 0.9 to 1.3, and exemplarily from 1.04 to 1.1.

[0029] The present application also provides a preparation method of the compound of formula A, comprising: reacting a compound of formula (I) with an appropriate amount of acid X to obtain the compound of formula A;

[0030] The n and X have the definitions as shown above.

[0031] According to an embodiment of the present application, the reaction is carried out in an organic solvent, for example, the organic solvent is selected from one or more than two of ethanol, acetone, 1,4-dioxane, n-heptane.

[0032] According to an embodiment of the present application, the temperature of the reaction is from 40 to 60°C, for example, 45°C, 50°C or 55°C.

[0033] The present application also provides a crystalline form of the compound of Formula A-I having X-ray powder diffraction peaks expressed in angles 2θ using Cu-Kα radiation at 8.1 ± 0.20°, 8.4 ± 0.20°, 13.4 ± 0.20°, 21.0 ± 0.20°, and 22.7 ± 0.20°; further having peaks at 5.4 ± 0.20°, 6.9 ± 0.20°, 9.1 ± 0.20°, 13.9 ± 0.20°, 18.0 ± 0.20°, 18.9 ± 0.20°, and / or 24.6 ± 0.20°; further having peaks at 11.0 ± 0.20°, 11.2 ± 0.20°, 14.5 ± 0.20°, 16.6 ± 0.20°, 17.4 ± 0.20°, 18.4 ± 0.20°, 20.0 ± 0.20°, 21.9 ± 0.20°, 23.2 ± 0.20°, 23.5 ± 0.20°, 25.5 ± 0.20°, 25.9 ± 0.20°, 26.2 ± 0.20°, 26.6 ± 0.20°, 27.0 ± 0.20°, 27.7 ± 0.20°, 28.0 ± 0.20°, 29.4 ± 0.20°, 30.1 ± 0.20°, and / or 30.5 ± 0.20°;

[0034] The compound of Formula A-I has the following structure:

[0035] n is selected from a number from 0.6 to 1.8, preferably from a number from 0.9 to 1.3, and is exemplarily selected from a number from 0.9 to 1.

[0036] According to an embodiment of the present application, the crystalline form of the compound of Formula A-I has an XRPD pattern substantially as shown in Figure 13.

[0037] According to an embodiment of the present application, the crystalline form of the compound of Formula A-I is an anhydrate.

[0038] According to an exemplary embodiment of the present application, the crystalline form of the compound of Formula A-I has a TGA pattern substantially as shown in Figure 15.

[0039] According to an embodiment of the present application, the crystalline form of the compound of Formula A-I has an endothermic peak with an onset temperature from 172 to 185 °C and a peak temperature difference from the onset temperature from 1 to 5 °C; for example, an endothermic peak with an onset temperature of 179.49 °C and a peak temperature difference from the onset temperature of 3.05 °C (peak temperature of 182.54 °C).

[0040] According to an exemplary embodiment of the present application, the crystalline form of the compound of Formula A-I has a DSC pattern substantially as shown in Figure 14.

[0041] The present application also provides a crystalline form of the compound of Formula A-II having an X-ray powder diffraction pattern, expressed in terms of 2 theta angles, using Cu-Ka radiation, having characteristic peaks at 8.2 ± 0.20°, 8.7 ± 0.20°, 13.7 ± 0.20°, 19.4 ± 0.20°, 21.3 ± 0.20°, and 23.1 ± 0.20°; further, can also have characteristic peaks at 5.5 ± 0.20°, 7.0 ± 0.20°, 9.4 ± 0.20°, 18.2 ± 0.20°, and / or 25.1 ± 0.20°; still further, can also have characteristic peaks at 10.8 ± 0.20°, 11.3 ± 0.20°, 14.1 ± 0.20°, 16.9 ± 0.20°, 18.9 ± 0.20°, 19.8 ± 0.20°, 20.6 ± 0.20°, 22.1 ± 0.20°, 22.4 ± 0.20°, 22.7 ± 0.20°, 25.7 ± 0.20°, 26.2 ± 0.20°, 27.8 ± 0.20°, 28.3 ± 0.20°, 30.0 ± 0.20°, 30.7 ± 0.20°, 31.6 ± 0.20°, 32.3 ± 0.20°, 32.5 ± 0.20°, and / or 32.7 ± 0.20°;

[0042] The compound of Formula A-II has the following structure:

[0043] n is selected from a number from 0.6 to 1.8, preferably a number from 0.9 to 1.3, and is illustratively selected from a number from 0.9 to 1.1 (e.g., 1).

[0044] According to an embodiment of the present application, the crystalline form of the compound of Formula A-II has an XRPD pattern substantially as shown in FIG. 19.

[0045] According to an embodiment of the present application, the crystalline form of the compound of Formula A-II is an anhydrate. According to an embodiment of the present application, the crystalline form of the compound of Formula A-II has a TGA pattern substantially as shown in FIG. 21.

[0046] According to an embodiment of the present application, the crystalline form of the compound of Formula A-II has an endothermic peak with an onset temperature from 175 to 185 °C and a peak temperature difference from the onset temperature from 1 to 8 °C; for example, an endothermic peak with an onset temperature of 181.42 °C and a peak temperature difference from the onset temperature of 3.89 °C (peak temperature of 185.31 °C).

[0047] According to an illustrative embodiment of the present application, the crystalline form of the compound of Formula A-II has a DSC pattern substantially as shown in FIG. 20.

[0048] The present application also provides a crystalline form of the compound of Formula A-III, which has characteristic peaks in the X-ray powder diffraction pattern, expressed in angles of 2-theta, using Cu-Kalpharadiation at 8.3±0.20°, 9.0±0.20°, 13.9±0.20°, 21.3±0.20°, 23.1±0.20° and 23.7±0.20°; further, it can also have characteristic peaks at 9.6±0.20°, 11.5±0.20°, 14.8±0.20°, 18.3±0.20° and / or 19.6±0.20°;

[0049] The compound of Formula A-III has the following structure:

[0050] n is selected from a number of 0.5 to 1.5, preferably a number of 0.9 to 1.3, and is exemplified by 1.

[0051] According to an embodiment of the present application, the crystalline form of the compound of Formula A-III has an XRPD pattern substantially as shown in Figure 28.

[0052] According to an embodiment of the present application, the crystalline form of the compound of Formula A-III is an anhydrate.

[0053] According to an embodiment of the present application, the crystalline form of the compound of Formula A-III has a weight loss of not more than 1 wt%, preferably not more than 0.8 wt%, and more preferably not more than 0.5 wt% at room temperature to 130°C.

[0054] According to an exemplary embodiment of the present application, the crystalline form of the compound of Formula A-III has a TGA pattern substantially as shown in Figure 30.

[0055] According to an embodiment of the present application, the crystalline form of the compound of Formula A-III has an endothermic peak with an onset temperature of 168 to 178°C and a difference between the peak temperature and the onset temperature of 1 to 6°C; for example, an endothermic peak with an onset temperature of 172.87°C and a difference between the peak temperature and the onset temperature of 3.27°C (peak temperature of 176.14°C).

[0056] According to an exemplary embodiment of the present application, the crystalline form of the compound of Formula A-III has a DSC pattern substantially as shown in Figure 29.

[0057] The present application also provides a crystal form of the compound of formula A-IV, which has characteristic peaks expressed in angles of 2θ using Cu-Kα radiation at 8.7±0.20°, 14.5±0.20°, 19.0±0.20°, 22.1±0.20°, 22.7±0.20° and 24.0±0.20°; further, it can also have characteristic peaks at 7.3±0.20°, 9.5±0.20°, 11.9±0.20°, 20.5±0.20°, 21.1±0.20°, 23.3±0.20° and / or 31.2±0.20°; still further, it can also have characteristic peaks at 10.0±0.20°, 26.2±0.20°, 29.3±0.20° and / or 29.7±0.20°;

[0058] The compound of formula A-IV has the following structure:

[0059] n is selected from a number of 0.5 to 1.5, preferably a number of 0.9 to 1.3, and exemplarily a number of 1.04 to 1.1.

[0060] According to an embodiment of the present application, the crystal form of the compound of formula A-IV has an XRPD pattern substantially as shown in Figure 31.

[0061] According to an embodiment of the present application, the crystal form of the compound of formula A-IV is an anhydrate.

[0062] According to an embodiment of the present application, the crystal form of the compound of formula A-IV has a weight loss of no more than 0.8wt%, preferably no more than 0.5wt%, and more preferably no more than 0.3wt% at room temperature to 130°C.

[0063] According to an exemplary embodiment of the present application, the crystal form of the compound of formula A-IV has a TGA pattern substantially as shown in Figure 33.

[0064] According to an embodiment of the present application, the crystal form of the compound of formula A-IV has an endothermic peak with an onset temperature of 162 to 172°C and a peak temperature difference from the onset temperature of 1 to 6°C; for example, an endothermic peak with an onset temperature of 168.00°C and a peak temperature difference from the onset temperature of 3.90°C (peak temperature of 171.90°C).

[0065] According to an exemplary embodiment of the present application, the crystal form of the compound of formula A-IV has a DSC pattern substantially as shown in Figure 32.

[0066] The present application also provides a pharmaceutical composition comprising one or two or more of the above-mentioned compound of formula A, the crystal form of the compound of formula A-I, the crystal form of the compound of formula A-II, the crystal form of the compound of formula A-III and the crystal form of the compound of formula A-IV.

[0067] According to embodiments of the present application, the pharmaceutical composition further comprises one, two or more pharmaceutically acceptable excipients.

[0068] According to embodiments of the present application, the pharmaceutical composition can further comprise an additional therapeutic agent for use in combination with the compound of Formula A.

[0069] According to embodiments of the present application, the additional therapeutic agent is selected from semaglutide.

[0070] The present application also provides the use of the compound of Formula A, the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, the crystalline form of the compound of Formula A-IV, or the pharmaceutical composition described above in the manufacture of a medicament for the treatment of a NLRP3-mediated condition and / or disease, such as in the manufacture of a NLRP3 inhibitor.

[0071] According to embodiments of the present application, the pharmaceutical composition comprises one or more of the compound of Formula A, the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, and the crystalline form of the compound of Formula A-IV, and semaglutide.

[0072] According to embodiments of the present application, the condition and / or disease is, for example, autoinflammatory fever syndromes such as cryopyrin-associated periodic syndromes (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), neuroinflammation-related disorders (e.g. multiple sclerosis, brain infections, acute injuries, neurodegenerative diseases, Alzheimer’s disease), atherosclerosis and cardiovascular risks (e.g. hypertension), obesity, hidradenitis suppurativa, wound healing and scarring, and cancer (e.g. colorectal cancer, lung cancer, myeloproliferative neoplasms, leukemia, myelofibrosis).

[0073] According to embodiments of the present application, the compound of Formula A, the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, and the crystalline form of the compound of Formula A-IV, or the pharmaceutical composition described above is used in the manufacture of a medicament for the treatment of obesity mediated by NLRP3.

[0074] The present application also provides a method for treating a NLRP3-mediated condition and / or disease, comprising administering to a patient a prophylactically or therapeutically effective amount of the compound of Formula A, the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, the crystalline form of the compound of Formula A-IV, or the pharmaceutical composition described above.

[0075] According to an embodiment of the present application, the pharmaceutical composition comprises one or two or more of the compound of Formula A, the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, and the crystalline form of the compound of Formula A-IV, and semaglutide.

[0076] According to an embodiment of the present application, the NLRP3-mediated condition and / or disease is selected from the group consisting of autoinflammatory fever syndromes such as cryopyrin-associated periodic syndromes (CAPS), sickle cell disease, systemic lupus erythematosus (SLE), chronic liver disease, nonalcoholic steatohepatitis (NASH), gout, gouty arthritis, pericarditis, type I and type II diabetes and related complications (e.g. nephropathy, retinopathy), neuroinflammation-related disorders (e.g. multiple sclerosis, brain infections, acute injury, neurodegenerative diseases, Alzheimer’s disease), atherosclerosis and cardiovascular risk (e.g. hypertension), obesity, hidradenitis suppurativa, wound healing and scarring, and cancer (e.g. colorectal cancer, lung cancer, myeloproliferative neoplasms, leukemia, myelofibrosis).

[0077] In some embodiments, the present application also provides a method for treating obesity, comprising administering to a patient a prophylactically or therapeutically effective amount of the compound of Formula A, the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, the crystalline form of the compound of Formula A-IV, or the pharmaceutical composition described above.

[0078] According to an embodiment of the present application, the method comprises administering to a patient a prophylactically or therapeutically effective amount of one or two or more of the compound of Formula A, the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, and the crystalline form of the compound of Formula A-IV, and semaglutide. Beneficial effects

[0079] The compound of Formula A has good biological activity and solubility, and has the potential to be developed into a drug.

[0080] The crystalline form of the compound of Formula A (e.g. the crystalline form of the compound of Formula A-I, the crystalline form of the compound of Formula A-II, the crystalline form of the compound of Formula A-III, the crystalline form of the compound of Formula A-IV) has low hygroscopicity, suitable solubility, and stability (including physical stability and chemical stability).

[0081] Definitions and explanations of terms

[0082] Unless otherwise stated, numerical ranges expressed in the specification and claims encompass every specific number within the range. For example, a range of 0.1 to 4 encompasses every number, e.g., 0.1, 0.2, 0.3, 0.4, 1, 2, 3, 4, etc. within the range.

[0083] The term "pharmaceutically acceptable excipient" means an excipient that is not biologically or otherwise undesirable, i.e., the excipient can be administered to an organism without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained.

[0084] The term "more than two" means two, three, four, five or more.

[0085] The term "patient" means any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, most preferably humans.

[0086] The term "therapeutically effective amount" means the amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought in a tissue, system, animal, individual or human by a researcher, veterinarian, medical doctor or other clinician, and includes one or more of the following: (1) preventing the disease: for example, preventing a disease, disorder or condition from occurring in an individual that is predisposed to the disease, disorder and / or condition but has not yet progressed to a pathologic state or symptoms of the disease; (2) inhibiting the disease: for example, arresting the development of a disease, disorder or condition (i.e., retarding the pathologic state and / or symptoms) in an individual that is experiencing or has experienced a pathologic state or symptoms of the disease, disorder or condition; (3) relieving the disease: for example, relieving a disease, disorder or condition in an individual that is experiencing or has experienced a pathologic state or symptoms of the disease, disorder or condition (i.e., reversing the pathologic state and / or symptoms).

[0087] The term "work-up" is an operation commonly used in the art to isolate a product, for example, including filtration, lyophilization, centrifugation and / or drying. BRIEF DESCRIPTION OF DRAWINGS

[0088] Figure 1 is a comparison of the results of circumference measurements of the rat ankle joint in a rat model of acute gouty arthritis induced by MSU.

[0089] Figure 2 is an XRPD pattern of the hydrochloride salt type A of the compound of formula (I).

[0090] Figure 3 is an XRPD pattern of the hydrochloride salt type B of the compound of formula (I).

[0091] Figure 4 is an XRPD pattern of the sulfate salt amorphous of the compound of formula (I).

[0092] Figure 5 is an XRPD pattern of the phosphate salt type A of the compound of formula (I).

[0093] Figure 6 is an XRPD pattern of the phosphate salt type B of the compound of formula (I).

[0094] Figure 7 is an XRPD pattern of the phosphate salt type C of the compound of formula (I).

[0095] Figure 8 is an XRPD pattern of the methanesulfonic acid salt type A of the compound of Formula (I).

[0096] Figure 9 is an XRPD pattern of the methanesulfonic acid salt type B of the compound of Formula (I).

[0097] Figure 10 is an XRPD pattern of the hydrobromide salt type A of the compound of Formula (I).

[0098] Figure 11 is an XRPD pattern of the hydrobromide salt type B of the compound of Formula (I).

[0099] Figure 12 is an XRPD pattern of the hydrobromide salt type C of the compound of Formula (I).

[0100] Figure 13 is an XRPD pattern of the maleic acid salt type A of the compound of Formula (I).

[0101] Figure 14 is a DSC pattern of the maleic acid salt type A of the compound of Formula (I).

[0102] Figure 15 is a TGA pattern of the maleic acid salt type A of the compound of Formula (I).

[0103] Figure 16 is an XRPD pattern of the maleic acid salt type B of the compound of Formula (I).

[0104] Figure 17 is an XRPD pattern of the maleic acid salt type C of the compound of Formula (I).

[0105] Figure 18 is an XRPD pattern of the maleic acid salt type D of the compound of Formula (I).

[0106] Figure 19 is an XRPD pattern of the maleic acid salt type E of the compound of Formula (I).

[0107] Figure 20 is a DSC pattern of the maleic acid salt type E of the compound of Formula (I).

[0108] Figure 21 is a TGA pattern of the maleic acid salt type E of the compound of Formula (I).

[0109] Figure 22 is an XRPD pattern of the fumaric acid salt type A of the compound of Formula (I).

[0110] Figure 23 is an XRPD pattern of the fumaric acid salt type B of the compound of Formula (I).

[0111] Figure 24 is an XRPD pattern of the L-malic acid salt type A of the compound of Formula (I).

[0112] Figure 25 is an XRPD pattern of L-malate type B of the compound of Formula (I).

[0113] Figure 26 is an XRPD pattern of L-tartrate type A of the compound of Formula (I).

[0114] Figure 27 is an XRPD pattern of L-tartrate type B of the compound of Formula (I).

[0115] Figure 28 is an XRPD pattern of malonate type A of the compound of Formula (I).

[0116] Figure 29 is a DSC pattern of malonate type A of the compound of Formula (I).

[0117] Figure 30 is a TGA pattern of malonate type A of the compound of Formula (I).

[0118] Figure 31 is an XRPD pattern of malonate type B of the compound of Formula (I).

[0119] Figure 32 is a DSC pattern of malonate type B of the compound of Formula (I).

[0120] Figure 33 is a TGA pattern of malonate type B of the compound of Formula (I).

[0121] Figure 34 is an XRPD pattern of p-toluenesulfonate type A of the compound of Formula (I).

[0122] Figure 35 is a DVS test pattern of maleate type A of the compound of Formula (I).

[0123] Figure 36 is a DVS test pattern of malonate type A of the compound of Formula (I).

[0124] Figure 37 is a DVS test pattern of malonate type B of the compound of Formula (I).

[0125] Figure 38 is the effect of the compound of Formula (I), semaglutide and their combination on the body weight of mice. DETAILED DESCRIPTION

[0126] The technical solutions of the present application will be further described in detail below in combination with specific examples. It should be understood that the following examples are only illustratively and explain the present application, and should not be interpreted as limiting the scope of protection of the present application. Any technology realized based on the above description of the present application is covered within the scope of the present application.

[0127] 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.

[0128] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0129] Nuclear magnetic resonance analysis (NMR) 1 H NMR)

[0130] 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).

[0131] Ion chromatography (IC)

[0132] The ion chromatograph was an ICS-6000 (Thermo, US), and its parameters are shown in Table A.

[0133] Table A IC Test Parameters

[0134] The maleate type A of the compound of formula (I) obtained in Example 27 and the maleate type E of the compound of formula (I) obtained in Example 31 were tested for NMR, XRPD, TGA and DSC using the following instruments and methods:

[0135] Hydrogen nuclear magnetic resonance (HNMR) 1 H-NMR)

[0136] Data was acquired using a Bruker 400MHz instrument. 1 H-NMR spectra. Unless otherwise specified, samples were prepared in CD3OD (deuterated methanol) solvent and tested according to the parameters in Table B-1. Data were analyzed using MestReNova software.

[0137] Table B-1 1 H-NMR analysis parameters

[0138] X-ray powder diffraction (XRPD)

[0139] 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 B-2.

[0140] Table B-2 Parameters for XRPD testing using Bruker D8 Advance

[0141] Thermogravimetric analysis (TGA)

[0142] Thermogravimetric analysis (TGA) was performed using a TA instrument. About 1-3 mg of sample was loaded into a pre-weighed aluminum pan and heated according to the parameters in Table B-3. Data were analyzed using TRIOS software.

[0143] Table B-3 Thermogravimetric analysis test parameters

[0144] Differential scanning calorimetry (DSC)

[0145] Differential scanning calorimetry (DSC) analysis was performed using a TA instrument. About 1-3 mg of sample was placed in an aluminum pan with a pinhole and heated according to the parameters in Table B-4. Data were analyzed using TRIOS software.

[0146] Table B-4 DSC test parameters

[0147] Except for the maleate salt type A and maleate salt type E of the compound of formula (I) obtained in Examples 27, 31, the following instruments and methods were used for XRPD, TGA, DSC tests of other examples:

[0148] X-ray powder diffraction (XRPD)

[0149] The solid samples obtained in the experiments were analyzed by X-ray powder diffractometer ARL Equinox 100. The 2 theta scanning angle was from 0° to 35°. The test method was Cu target K alpha ray, voltage 40 kV, current 0.9 mA, and the sample pan was a zero background sample pan.

[0150] Thermogravimetric analysis (TGA)

[0151] The thermogravimetric analyzer was a METTLER TOLEDO TGA2 (METTLER TOLEDO, US). 2-10 mg of sample was placed in an open aluminum sample pan that had been equilibrated and automatically weighed in the TGA furnace. The sample was heated at a rate of 10 °C / min to the final temperature, and the sample was purged with nitrogen at a rate of 50 mL / min and the balance was purged with nitrogen at a rate of 20 mL / min.

[0152] Differential scanning calorimetry analysis (DSC)

[0153] The model of the differential scanning calorimeter is METTLER TOLEDO DSC3 (METTLER TOLEDO, US). 2-10 mg of sample was accurately weighed and placed in a pierced DSC Tzero sample pan, heated to 80℃ or 100℃ at a rate of 2℃ / min, held for 5 min, then decreased to 25℃ at a rate of 10℃ / min, held for 10 min at 25℃, increased to the final temperature at a rate of 2℃ / min, and the nitrogen purge rate in the furnace was 50 mL / min.

[0154] Example 1

[0155] First step: Preparation of (S)-3-((7-bromo-4-chlorophthalazin-1-yl)amino)propane-1,2-diol (I-B)

[0156] Compound I-A (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 heated to 80℃ for 4 hours. After the reaction was completed, the reaction liquid was concentrated and the obtained residue was purified by silica gel column chromatography with a eluent of dichloromethane / methanol system to obtain the title compound I-B (1.2 g, yield: 25%).

[0157] MS m / z (ESI): 332.1 (M+1).

[0158] Second step: Preparation of (S)-3-((4-chloro-7-(prop-1-yn-1-yl)phthalazin-1-yl)amino)propane-1,2-diol (I-C)

[0159] Compound I-B (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 dichlorobis(triphenylphosphine)palladium (278 mg, 0.39 mmol) were dissolved in a mixed solvent of N,N-dimethylacetamide and water (22 mL, V / V=10:1), and heated to 100℃ for 4 hours. After the reaction was completed, the reaction liquid was added to a saturated ammonium chloride solution, extracted with ethyl acetate (10 mL x 3), filtered and concentrated to obtain a crude product. The obtained residue was purified by silica gel column chromatography with a eluent of dichloromethane / methanol to obtain the title compound I-C (70 mg, yield: 13.5%).

[0160] MS m / z (ESI): 292.1 (M+1).

[0161] 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)

[0162] Compound I-C (50 mg, 0.17 mmol) was dissolved in a mixed solvent of dioxane and water (2.2 mL, V / V = 10:1), (4-chloro-2-hydroxyphenyl)boronic acid (30 mg, 0.17 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (12 mg, 0.02 mmol) and potassium carbonate (47 mg, 0.34 mmol) were added, and the reaction solution 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 preparation (Waters MS triggered Prep-LC with Acquity QDA detector, column: Welch 10 μm C18 250 x 21.2 mm; mobile phase 1: water (containing 0.1% NH3); mobile phase 2: acetonitrile; 15 minute gradient, gradient ratio: acetonitrile phase 15%-100%, flow rate: 25 mL / min) to obtain compound I (also referred to as compound 062), i.e. a compound of formula (I) (4.4 mg, yield: 6.7%).

[0163] MS m / z (ESI): 384.1 (M+1) + .

[0164] 1 H NMR (400 MHz, CD3OD) δ 8.25 (s, 1H), 7.72 (dd, 1H), 7.55 (d, 1H), 7.29 (d, 1H), 7.01 (d, 1H), 6.99 (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).

[0165] Example 2

[0166] Compound 40 mg of formula (I) was added to 0.4 ml of ethanol, 125 μl of 1M hydrochloric acid ethanol solution was added, and after stirring at 50 °C for 3 hours, it was concentrated, then a certain amount of water (about 50V) was added to dissolve it, and freeze-drying was performed to obtain compound of formula A-1.

[0167] 1H NMR (400 MHz, DMSO-d6) δ 10.68 (s, 1H), 10.00 (s, 1H), 8.94 (s, 1H), 8.01 (dd, J = 8.5, 1.4 Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.37 (d, J = 8.1 Hz, 1H), 7.21 - 7.03 (m, 2H), 5.32 (t, J = 4.8 Hz, 1H), 3.92 (dt, J = 9.6, 4.9 Hz, 1H), 3.75 (dt, J = 13.9, 4.7 Hz, 1H), 3.67 (s, 1H), 3.48 (dq, J = 11.0, 5.9 Hz, 3H), 2.17 (s, 3H), 1.36 - 1.15 (m, 1H).

[0168] The signals at 6.5 ppm to 9.0 ppm and so on are shifted compared with the free state, indicating that the sample is successfully prepared.

[0169] The IC test result shows that the content of chloride ion is 8.2%, that is, n = 1.1 in the compound of formula A-1.

[0170] Example 3

[0171] The compound of formula (I) 40 mg was added to 0.4 ml of ethanol, 125 μl of 1M sulfuric acid ethanol solution was added, and after stirring at 50°C for 3 hours, it was cooled to room temperature, and stirred at room temperature for 1 day, centrifuged, and the solid was dried at 50°C under vacuum to obtain the compound of formula A-2.

[0172] 1 H NMR (400 MHz, DMSO-d6) δ 10.57 (s, 1H), 8.84 (s, 1H), 8.01 (dd, J = 8.5, 1.4 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.37 (d, J = 8.0 Hz, 1H), 7.15 - 7.04 (m, 2H), 3.90 (dd, J = 9.1, 5.0 Hz, 2H), 3.78 - 3.69 (m, 2H), 3.63 (dd, J = 14.2, 7.2 Hz, 3H), 3.55 - 3.40 (m, 6H), 2.17 (s, 3H), 2.00 (q, J = 6.9, 6.4 Hz, 1H), 1.24 (s, 3H), 1.05 (t, J = 7.0 Hz, 2H), 0.90 - 0.81 (m, 1H).

[0173] The signals at 6.5 ppm to 9.0 ppm and so on are shifted compared with the free state, indicating that the sample is successfully prepared.

[0174] IC test results showed the content of sulfate ions was 20.2%, i.e. n = 1.05 in the compound of formula A-2.

[0175] The product obtained was characterized by XRPD, which showed that it was amorphous (see Figure 4). TGA test results showed that the product lost about 2.5wt% at room temperature to 60°C, and lost about 9.0wt% at 60-140°C.

[0176] Example 4

[0177] The compound of formula (I) 40 mg was added to 0.2 ml of ethanol, 125 μl of 1 M ethanolic methanesulfonic acid solution was added, stirred at 50°C for 3 hours, then cooled to room temperature, stirred overnight, 1.2 ml of n-heptane was added and stirred for a certain period of time, then concentrated, and then a certain amount of water (about 50V) was added to dissolve, and then freeze-dried to obtain the compound of formula A-4.

[0178] 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (d, J = 1.5 Hz, 1H), 7.74 (dd, J = 8.4, 1.5 Hz, 1H), 7.67 (s, 1H), 7.40 (d, J = 8.5 Hz, 1H), 7.33 - 7.25 (m, 1H), 7.07 - 6.96 (m, 2H), 3.90 - 3.80 (m, 2H), 3.69 (d, J = 13.5 Hz, 2H), 3.53 (dd, J = 13.5, 6.8 Hz, 2H), 3.42 (s, 3H), 2.13 (s, 3H), 2.04 - 1.97 (m, 1H), 1.30 - 1.17 (m, 2H).

[0179] Compared with the free state, the signals at 6.5 ppm to 9.0 ppm and other signals were shifted, indicating that the sample was successfully prepared.

[0180] IC test results showed the content of phosphate ions was 25.7%, i.e. n = 1.45 in the compound of formula A-3.

[0181] Example 5

[0182] The compound of formula (I) 40 mg was added to 0.2 ml of ethanol, 125 μl of 1 M ethanolic methanesulfonic acid solution was added, stirred at 50°C for 3 hours, then cooled to room temperature, stirred overnight, 1.2 ml of n-heptane was added and stirred for a certain period of time, then concentrated, and then a certain amount of water (about 50V) was added to dissolve, and then freeze-dried to obtain the compound of formula A-4.

[0183] 1H NMR (400 MHz, DMSO-d6) δ 10.58 (s, 1H), 9.83 (s, 1H), 8.85 (d, J = 1.5 Hz, 1H), 8.02 (dd, J = 8.4, 1.5 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.38 (d, J = 8.0 Hz, 1H), 7.10 (d, J = 7.7 Hz, 2H), 4.02 - 3.87 (m, 1H), 3.73 (dt, J = 13.9, 4.7 Hz, 1H), 3.65 (d, J = 7.2 Hz, 1H), 3.49 (qd, J = 11.0, 5.5 Hz, 5H), 2.30 (s, 3H), 2.13 (s, 3H), 1.35 - 1.16 (m, 2H), 0.91 - 0.79 (m, 1H).

[0184] Compared with the free state, the signal peaks at 6.5 ppm to 9.0 ppm and other multiple signal peaks are shifted, and the signal peak of methyl sulfonic acid at 2.3 ppm can be seen, indicating that the sample is successfully prepared. According to the nuclear magnetic integration result, it is calculated that n = 1.05 in the compound of formula A-4.

[0185] Example 6

[0186] The compound of formula (I) 40 mg was added to 0.2 ml of acetone, 125 μl of 1M hydrobromic acid acetone solution was added, and after stirring at 50°C for 3 hours, it was cooled to room temperature, and stirred overnight, centrifuged, and the solid was dried at 50°C under vacuum to obtain the compound of formula A-5.

[0187] 1 H NMR (400 MHz, DMSO-d6) δ 10.58 (s, 1H), 9.83 (s, 1H), 8.85 (d, J = 1.5 Hz, 1H), 8.02 (dd, J = 8.4, 1.5 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.38 (d, J = 8.0 Hz, 1H), 7.10 (d, J = 7.7 Hz, 2H), 4.02 - 3.87 (m, 1H), 3.73 (dt, J = 13.9, 4.7 Hz, 1H), 3.65 (d, J = 7.2 Hz, 1H), 3.49 (qd, J = 11.0, 5.5 Hz, 5H), 2.30 (s, 3H), 2.13 (s, 3H), 1.35 - 1.16 (m, 2H), 0.91 - 0.79 (m, 1H).

[0188] Compared with the free state, the signal peaks at 6.5 ppm to 9.0 ppm and other multiple signal peaks are shifted, indicating that the compound is successfully prepared.

[0189] The IC result shows that the content of bromide ion is 22.1%, and it is calculated that n = 1.43 in the compound of formula A-5.

[0190] Example 7

[0191] Compound of formula (I) 40 mg was added to 0.2 ml 1,4-dioxane, 125 μl of 1M hydrobromic acid 1,4-dioxane solution was added, stirred at 50°C for 3 hours, then cooled to room temperature, stirred overnight, added 0.6 ml n-heptane, continued to stir at room temperature, centrifuged, the solid was dried at 50°C under vacuum to obtain compound of formula A-6.

[0192] 1 H NMR (400 MHz, DMSO-d6) δ 10.56 (s, 1H), 9.81 (s, 1H), 8.85 (d, J = 1.5 Hz, 1H), 8.02 (dd, J = 8.4, 1.5 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.38 (d, J = 8.0 Hz, 1H), 7.10 (d, J = 7.7 Hz, 2H), 3.99 - 3.86 (m, 1H), 3.73 (dt, J = 13.9, 4.8 Hz, 1H), 3.65 (d, J = 7.1 Hz, 1H), 3.56 (s, 2H), 3.49 (qd, J = 11.1, 5.6 Hz, 3H), 2.17 (s, 3H), 2.00 (q, J = 6.8, 6.4 Hz, 1H), 1.23 (s, 2H).

[0193] The signals of 6.5 ppm to 9.0 ppm and other signals were shifted compared with the free state, indicating that the compound was successfully prepared. The structure of compound of formula A-6 was the same as that of compound of formula A-5, and the IC result showed that the content of bromide ion was 19.3%, and n = 1.21 was calculated in the compound of formula A-6.

[0194] Example 8

[0195] Compound of formula (I) 40 mg was added to 0.8 ml ethanol, 14.7 mg maleic acid was added, stirred at 50°C for 3 hours, then concentrated, added a certain amount of water (about 50V) to dissolve, freeze-dried to obtain compound of formula A-7.

[0196] 1H NMR (400 MHz, DMSO-d6) δ 10.45 (s, 1H), 8.72 (s, 1H), 7.92 (d, J = 8.5 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.35 (d, J = 8.0 Hz, 1H), 7.07 (d, J = 7.5 Hz, 2H), 6.10 (s, 2H), 3.90 (q, J = 5.2 Hz, 1H), 3.71 (dt, J = 13.9, 4.8 Hz, 1H), 3.60 (dt, J = 14.0, 6.8 Hz, 1H), 3.47 (h, J = 5.2 Hz, 2H), 2.16 (s, 3H), 2.00 (q, J = 7.0, 6.6 Hz, 1H), 1.45 (s, 1H), 1.34 - 1.15 (m, 3H), 0.92 - 0.79 (m, 1H).

[0197] Compared with the free state, the signal peaks at 6.5 ppm to 9.0 ppm are shifted, and the signal peak of maleic acid at 6.1 ppm can be seen, indicating that the compound is successfully prepared. According to the integral result of nuclear magnetic resonance, it is calculated that n = 1.0 in the compound of formula A-7.

[0198] Example 9

[0199] The compound of formula (I) 40 mg was added to 0.8 ml of ethanol, 14.7 mg of fumaric acid was added, and after stirring at 50°C for 3 hours, it was cooled to room temperature, stirred overnight, centrifuged, and the solid was dried at 50°C under vacuum to obtain the compound of formula A-8.

[0200] 1 H NMR (400 MHz, Methanol-d4) δ 9.27 (d, J = 1.5 Hz, 1H), 8.54 (dd, J = 8.5, 1.5 Hz, 1H), 8.44 (s, 1H), 8.21 (d, J = 8.5 Hz, 1H), 8.15 - 8.07 (m, 1H), 7.82 (d, J = 6.9 Hz, 2H), 7.42 (s, 1H), 4.71 - 4.61 (m, 1H), 4.50 (d, J = 13.7 Hz, 1H), 4.39 - 4.29 (m, 1H), 4.23 (d, J = 5.4 Hz, 2H), 2.94 (s, 3H), 2.86 - 2.72 (m, 1H), 2.26 (s, 1H), 2.15 - 1.96 (m, 2H), 1.72 - 1.61 (m, 1H).

[0201] Compared with the free state, the signal peaks at 6.5 ppm to 9.0 ppm and other places are shifted, and the signal peak of fumaric acid at about 7.4 ppm can be seen, indicating that the compound is successfully prepared. According to the nuclear magnetic integration result, it is calculated that n = 0.5 in the compound of formula A-8.

[0202] Example 10

[0203] The compound of formula (I) 40 mg is added to 0.4 ml of acetone, 14.7 mg of fumaric acid is added, and after stirring at 50°C for 3 hours, it is cooled to room temperature, stirred overnight, centrifuged, and the solid is vacuum dried at 50°C to obtain the compound of formula A-9.

[0204] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 1.5 Hz, 1H), 7.73 (dd, J = 8.4, 1.5 Hz, 1H), 7.63 (s, 1H), 7.40 (d, J = 8.5 Hz, 1H), 7.33 - 7.26 (m, 1H), 7.01 (d, J = 7.1 Hz, 2H), 6.62 (s, 2H), 3.91 - 3.80 (m, 1H), 3.69 (d, J = 14.0 Hz, 1H), 3.59 - 3.48 (m, 2H), 3.42 (d, J = 5.4 Hz, 3H), 2.13 (s, 3H), 1.28 - 1.19 (m, 1H).

[0205] Compared with the free state, the signal peaks at 6.5 ppm to 8.5 ppm and other places are shifted, and the signal peak of fumaric acid at 6.62 ppm can be seen, indicating that the compound is successfully prepared. The structural formula of the compound of formula A-9 is the same as that of the compound of formula A-8. According to the nuclear magnetic integration result, it is calculated that n = 0.8 in the compound of formula A-9.

[0206] Example 11

[0207] The compound of formula (I) 40 mg is added to 0.4 ml of ethanol, 16.9 mg of L-malic acid is added, and after stirring at 50°C for 3 hours, it is cooled to room temperature, stirred overnight, centrifuged, and the solid is vacuum dried at 50°C to obtain the compound of formula A-10.

[0208] 1H NMR (400 MHz, DMSO-d6) δ 10.19 (s, 1H), 8.47 (d, J = 1.5 Hz, 1H), 7.84 - 7.62 (m, 2H), 7.40 (d, J = 8.5 Hz, 1H), 7.34 - 7.24 (m, 1H), 7.10 - 6.95 (m, 2H), 4.22 (dd, J = 7.5, 5.2 Hz, 1H), 3.91 - 3.81 (m, 1H), 3.69 (dt, J = 13.5, 4.9 Hz, 1H), 3.52 (dt, J = 12.9, 5.5 Hz, 1H), 3.42 (d, J = 5.5 Hz, 2H), 2.60 (dd, J = 15.7, 5.2 Hz, 1H), 2.42 (dd, J = 15.6, 7.5 Hz, 1H), 2.13 (s, 3H), 2.00 (q, J = 7.1 Hz, 1H), 1.23 (s, 2H).

[0209] Compared with the free state, the signal peaks at 6.5 ppm to 8.5 ppm and other places are shifted, and the signal peaks of L-malic acid can be seen near 4.22 ppm, 2.6 ppm, 2.4 ppm, indicating that the compound is successfully prepared. According to the integral result of nuclear magnetic resonance, it is calculated that n = 1 in the compound of formula A-10.

[0210] Example 12

[0211] The compound 40 mg shown in formula (I) is added to 0.4 ml of ethanol, 19.0 mg of L-tartaric acid is added, stirred at 50°C for 3 hours, then reduced to room temperature, stirred overnight, centrifuged, and the solid is dried at 50°C under vacuum to obtain the compound of formula A-11.

[0212] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 1.6 Hz, 1H), 7.78 - 7.60 (m, 2H), 7.40 (d, J = 8.5 Hz, 1H), 7.33 - 7.25 (m, 1H), 7.07 - 6.96 (m, 2H), 4.27 (s, 1H), 3.92 - 3.80 (m, 1H), 3.69 (dt, J = 13.4, 5.0 Hz, 1H), 3.52 (dt, J = 12.8, 5.2 Hz, 1H), 2.13 (s, 3H), 2.04 - 1.94 (m, 1H), 1.23 (s, 2H).

[0213] Compared with the free state, the signal peaks at 6.5 ppm to 8.5 ppm and other places are shifted, and the signal peaks of L-malic acid can be seen near 4.22 ppm, 2.6 ppm, 2.4 ppm, indicating that the compound is successfully prepared. According to the integral result of nuclear magnetic resonance, it is calculated that n = 1 in the compound of formula A-10.

[0214] Example 13

[0215] The compound of formula (I) 40 mg was added to 0.4 ml of acetone, 19.0 mg of L-tartaric acid was added, stirred at 50 °C for 3 hours, then cooled to room temperature, stirred overnight, centrifuged, and the solid was dried at 50 °C under vacuum to obtain the compound of formula A-12.

[0216] 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (d, J = 1.6 Hz, 1H), 7.74 (dd, J = 8.5, 1.5 Hz, 1H), 7.66 (s, 1H), 7.40 (d, J = 8.5 Hz, 1H), 7.33 - 7.26 (m, 1H), 7.04 - 6.97 (m, 2H), 4.27 (s, 2H), 3.90 - 3.79 (m, 1H), 3.69 (d, J = 14.1 Hz, 1H), 3.59 - 3.47 (m, 2H), 3.42 (d, J = 5.5 Hz, 3H), 2.13 (s, 3H), 2.08 (s, 2H), 1.23 (s, 1H).

[0217] The signal peaks at 6.5 ppm to 8.5 ppm and other signal peaks were shifted compared with the free state, and the signal peak of L-tartaric acid was observed near 4.27 ppm, indicating that the compound was successfully prepared. According to the integral result of nuclear magnetic resonance, it was calculated that n = 1.1 in the compound of formula A-12.

[0218] Example 14

[0219] The compound of formula (I) 40 mg was added to 0.8 ml of ethanol, 13.4 mg of malonic acid was added, stirred at 50 °C for 3 hours, then concentrated, and then a certain amount of water (about 50V) was added to dissolve, freeze-dried to obtain the compound of formula A-13.

[0220] 1H NMR (400 MHz, DMSO-d6) δ 8.51 (s, 1H), 7.77 (dd, J = 8.4, 1.5 Hz, 2H), 7.42 (d, J = 8.5 Hz, 1H), 7.30 (d, J = 8.6 Hz, 1H), 7.10 - 6.95 (m, 2H), 3.85 (dt, J = 10.2, 5.2 Hz, 1H), 3.68 (dd, J = 11.6, 6.9 Hz, 1H), 3.60 - 3.49 (m, 1H), 3.42 (d, J = 5.5 Hz, 2H), 3.17 (s, 2H), 2.13 (s, 3H), 2.00 (q, J = 7.0, 6.5 Hz, 1H), 1.45 (s, 1H), 1.33 - 1.17 (m, 3H), 0.90 - 0.81 (m, 1H).

[0221] Compared with the free state, the signal peaks at 6.5 ppm to 8.5 ppm and other places are shifted, and the signal peak of malonic acid can be seen near 3.17 ppm, indicating that the compound is successfully prepared. According to the integral result of nuclear magnetic resonance, it is calculated that n = 1 in the compound of formula A-13.

[0222] Example 15

[0223] The compound of formula (I) 40 mg was added to 0.2 ml of ethanol, 24.3 mg of p-toluenesulfonic acid monohydrate was added, and after stirring at 50°C for 3 hours, it was cooled to room temperature, and 0.6 ml of n-heptane was added. Continue to stir at room temperature, and after post-treatment, the compound of formula A-14 is obtained.

[0224] 1 H NMR (400 MHz, DMSO-d6) δ 10.58 (s, 1H), 9.83 (s, 1H), 8.85 (d, J = 1.5 Hz, 1H), 8.02 (dd, J = 8.4, 1.5 Hz, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.51 - 7.43 (m, 2H), 7.38 (d, J = 8.0 Hz, 1H), 7.11 (d, J = 2.0 Hz, 1H), 7.10 - 7.07 (m, 2H), 3.92 (t, J = 5.7 Hz, 1H), 3.72 (dt, J = 13.9, 4.9 Hz, 1H), 3.65 (s, 1H), 3.56 - 3.41 (m, 7H), 2.28 (s, 3H), 2.17 (s, 3H), 1.23 (s, 1H).

[0225] Compared with the free state, the signal peaks at 6.5 ppm to 9.0 ppm and other places are shifted, and the signal peaks of p-toluenesulfonic acid can be seen near 7.46 ppm, 7.10 ppm and 2.28 ppm, indicating that the compound is successfully prepared. According to the integral result of nuclear magnetic resonance, it is calculated that n = 1 in the compound of formula A-14.

[0226] Example 16

[0227] The compound of formula (I) 40 mg was added to 0.4 ml of ethanol, 125 μl of 1M hydrochloric acid ethanol solution, stirred at 50°C for 3 hours, then cooled to room temperature, stirred for 1 day, centrifuged, and the solid was dried in a vacuum oven at 50°C to obtain the product.

[0228] The XRPD test results (Figure 2) of the product showed that it was a crystal form, and the XRPD analysis results are shown in Table 1. This crystal form is recorded as the hydrochloride type A of the compound of formula (I).

[0229] Table 1

[0230] Example 17

[0231] The compound of formula (I) 40 mg was added to 0.4 ml of ethanol, 125 μl of 1M hydrochloric acid ethanol solution, stirred at 50°C for 3 hours, then cooled to room temperature, stirred for 1 day, centrifuged, and the solid was dried in a vacuum oven at 50°C to obtain the product.

[0232] The XRPD test results (Figure 2) of the product showed that it was a crystal form, and the XRPD analysis results are shown in Table 1. This crystal form is recorded as the hydrochloride type A of the compound of formula (I).

[0233] Table 2

[0234] Example 18

[0235] The product obtained in Example 3 was characterized by XRPD and showed to be amorphous (see Figure 4).

[0236] Example 19

[0237] The compound of formula (I) 40 mg was added to 0.4 ml of ethanol, 125 μl of 1M hydrochloric acid ethanol solution, stirred at 50°C for 3 hours, then cooled to room temperature, stirred for 1 day, centrifuged, and the solid was dried in a vacuum oven at 50°C to obtain the product.

[0238] The XRPD test results (Figure 2) of the product showed that it was a crystal form, and the XRPD analysis results are shown in Table 1. This crystal form is recorded as the hydrochloride type A of the compound of formula (I).

[0239] Table 3

[0240] Example 20

[0241] The product obtained in Example 4 was characterized by XRPD, which showed to be a crystalline form (see Figure 6) and the XRPD pattern is shown in Table 4. This crystalline form was designated as phosphate salt type B of the compound of formula (I).

[0242] Table 4

[0243] Example 21

[0244] The compound of formula (I) 40 mg was added to 0.5 ml of 1,4-dioxane, 125 μL of 1 M phosphoric acid in 1,4-dioxane was added, after stirring at 50 °C for 3 hours it was allowed to cool to room temperature, stirred overnight, centrifuged, the solid was dried in a vacuum oven at 50 °C to obtain the product.

[0245] The XRPD test results of the product (Figure 7) showed that it was a crystalline form and the XRPD pattern is shown in Table 5. This crystalline form was designated as phosphate salt type C of the compound of formula (I).

[0246] Table 5

[0247] Example 22

[0248] The compound of formula (I) 40 mg was added to 0.4 ml of acetone, 229 μL of 1 M methanesulfonic acid in acetone was added, after stirring at 50 °C for 3 hours it was allowed to cool to room temperature, stirred overnight, centrifuged, the solid was dried in a vacuum oven at 50 °C to obtain the product.

[0249] The XRPD test results of the product (Figure 8) showed that it was a crystalline form and the XRPD pattern is shown in Table 6. This crystalline form was designated as methanesulfonic acid salt type A of the compound of formula (I).

[0250] Table 6

[0251] Example 23

[0252] The compound of formula (I) 40 mg was added to 0.2 ml of ethanol, 125 μL of 1 M methanesulfonic acid in ethanol was added, after stirring at 50 °C for 3 hours it was allowed to cool to room temperature, after stirring overnight, 1.2 ml of n-heptane was added, after stirring at room temperature for a certain time, centrifuged, the solid was dried in a vacuum oven at 50 °C to obtain the product.

[0253] The XRPD test result of the product (Figure 9) showed that it was a crystalline form, and the XRPD analysis result is shown in Table 7. This crystalline form is designated as the methanesulfonic acid salt type B of the compound of Formula (I).

[0254] Table 7

[0255] Example 24

[0256] The product obtained from Example 6 was characterized by XRPD, which showed that it was a crystalline form (see Figure 10), and the XRPD analysis result is shown in Table 8. This crystalline form is designated as the hydrobromide salt type A of the compound of Formula (I).

[0257] Table 8

[0258] Example 25

[0259] The compound of Formula (I) 40 mg was added with 0.2 ml of ethanol, 125 μL of 1M hydrobromic acid ethanol solution was added, after stirring at 50°C for 3 hours, it was cooled to room temperature, stirred overnight, 1.2 ml of n-heptane was added, continued to stir at room temperature for a certain period of time, centrifuged, the solid was dried in a vacuum oven at 50°C, to obtain the product.

[0260] The XRPD test result of the product (Figure 11) showed that it was a crystalline form, and the XRPD analysis result is shown in Table 9. This crystalline form is designated as the hydrobromide salt type B of the compound of Formula (I).

[0261] Table 9

[0262] Example 26

[0263] The product obtained from Example 7 was characterized by XRPD, which showed that it was a crystalline form (see Figure 12), and the XRPD analysis result is shown in Table 10. This crystalline form is designated as the hydrobromide salt type C of the compound of Formula (I).

[0264] Table 10

[0265] Example 27

[0266] The compound of Formula (I) 40 mg was added with 0.8 ml of ethanol, 14.7 mg of maleic acid was added, after stirring at 50°C for 3 hours, it was cooled to room temperature, stirred overnight, centrifuged, the solid was dried in a vacuum oven at 50°C, to obtain the compound of Formula A-I.

[0267] NMR test results showed that compared with free state, the signal peaks at 6.5 ppm to 9.0 ppm and other places were shifted, indicating that the sample was salted; the NMR integral results were basically consistent with the raw materials, and the signal peak of maleic acid was visible at 6.1 ppm, according to the integral results, the salted ratio was 1:0.9, that is, n was 0.9, and no obvious solvent peak was observed.

[0268] The compound of formula A-I was analyzed by XRPD (see Figure 13) and was found to be a crystalline form, which was designated as the maleate salt type A. The XRPD analysis results are shown in Table 11. The TGA test results showed that the product had almost no weight loss at room temperature to 170°C (Figure 15). The DSC showed that the product had an endothermic peak with an onset temperature of 179.49°C and a peak temperature of 182.56°C (Figure 14).

[0269] Table 11

[0270] Example 28

[0271] The compound of formula (I) 2g, 0.64g maleic acid, 64ml EtOH (ethanol) / n-heptane (normal heptane) = 2v / 1v were added, stirred at 30°C for 1 day, then reduced to 5°C, stirred, centrifuged, and the solid was dried in a vacuum oven at 50°C to obtain the product.

[0272] The XRPD test results (Figure 16) of the product showed that it was a crystalline form, and the XRPD analysis results are shown in Table 12. This crystalline form was designated as the maleate salt type B of the compound of formula (I).

[0273] Table 12

[0274] Example 29

[0275] The compound of formula (I) 30mg, 10.9mg maleic acid, 0.6ml NMP (N-methyl pyrrolidone) / IPAC (isopropyl acetate) = 1v / 7v were added, stirred at 50°C-5°C at a temperature rising and falling rate of 0.1°C / min for 7 days, then centrifuged at 5°C to obtain the product.

[0276] The XRPD test results (Figure 17) of the product showed that it was a crystalline form, and the XRPD analysis results are shown in Table 13. This crystalline form was designated as the maleate salt type C of the compound of formula (I).

[0277] Table 13

[0278] Example 30

[0279] The compound of formula (I) 30 mg, maleic acid 10.9 mg, MeOH (methanol) 0.4 mL were dissolved at room temperature and then placed in an acetone atmosphere for vapor diffusion to obtain the product.

[0280] The XRPD test result of the product (Figure 18) showed that it was a crystal form, and the XRPD analysis result is shown in Table 14. This crystal form was recorded as the maleate salt type D of the compound of formula (I).

[0281] Table 14

[0282] Example 31

[0283] The compound of formula (I) 100 mg, maleic acid 31.7 mg, anisole (anisole) / EtOH (ethanol) = 1 v / 9 v 2 ml were stirred at 50°C for 3 days, centrifuged and dried in a vacuum oven at 50°C to obtain the compound of formula A-II (n = 1).

[0284] The NMR test result showed that the signal peaks at 6.5 ppm to 9.0 ppm were shifted compared with the free state, indicating that the sample was salted; the NMR integral result was basically consistent with the raw material, and the signal peak of maleic acid at 6.1 ppm was observed, and the salted ratio was calculated to be 1:1 according to the integral result, that is, n was 1, and no obvious solvent peak was observed.

[0285] The XRPD analysis (see Figure 19) showed that the compound of formula A-II was a crystal form, which was recorded as the maleate salt type E, and the specific XRPD analysis spectrum is shown in Table 15. The TGA test result showed that the product had a weight loss of about 0.1% at room temperature to 170°C (Figure 21). DSC showed that the product had an endothermic peak with an onset temperature of 181.42°C and a peak temperature of 185.31°C (Figure 20).

[0286] Table 15

[0287] Example 32

[0288] The product obtained in Example 9 was characterized by XRPD and showed a crystal form (see Figure 22), and the XRPD analysis spectrum is shown in Table 16. This crystal form was recorded as the fumarate salt Type A of the compound of formula (I).

[0289] Table 16

[0290] Example 33

[0291] The product obtained in Example 10 was characterized by XRPD, which showed a crystalline form (see Figure 23), and the XRPD pattern is shown in Table 17. This crystalline form was designated as the fumarate salt Type B of the compound of Formula (I).

[0292] Table 17

[0293] Example 34

[0294] The product obtained in Example 11 was characterized by XRPD, which showed a crystalline form (see Figure 24), and the XRPD pattern is shown in Table 18. This crystalline form was designated as the L-malate salt type A of the compound of Formula (I).

[0295] Table 18

[0296] Example 35

[0297] The compound of Formula (I) 40 mg was added to 0.2 ml 1,4-dioxane, and 16.9 mg L-malic acid. After stirring at 50 °C for 3 hours, it was allowed to cool to room temperature, stirred overnight, and then warmed to 50 °C for 3 days. It was centrifuged, and the solid was dried in a vacuum oven at 50 °C to give the product.

[0298] The product was tested by XRPD (Figure 25), which showed a crystalline form, and the XRPD pattern is shown in Table 19. This crystalline form was designated as the L-malate salt type B of the compound of Formula (I).

[0299] Table 19

[0300] Example 36

[0301] The product obtained in Example 12 was characterized by XRPD, which showed a crystalline form (see Figure 26), and the XRPD pattern is shown in Table 20. This crystalline form was designated as the L-tartrate salt type A of the compound of Formula (I).

[0302] Table 20

[0303] Example 37

[0304] The product obtained in Example 13 was characterized by XRPD, which showed a crystalline form (see Figure 27), and the XRPD pattern is shown in Table 21. This crystalline form was designated as the L-tartrate salt type B of the compound of Formula (I).

[0305] Table 21

[0306] Example 38

[0307] The compound of formula (I) 40 mg was added with 0.6 ml 1,4-dioxane, and 13.4 mg malonic acid. After stirring at 50 °C for 3 hours, the temperature was decreased to room temperature, and the stirring was continued overnight. The solid was centrifuged and dried in a vacuum oven at 50 °C to obtain the compound of formula A-IV.

[0308] The NMR results showed that, compared with the free state, multiple signal peaks from 4.5 ppm to 5.5 ppm were shifted, indicating that the sample was salted. The NMR integral results were basically consistent with the raw material. The signal peak of malonic acid was observed near 3.16 ppm. According to the integral calculation, the salted ratio was 1:1.1 (i.e. n was 1.1). The solvent peak of 1,4-dioxane was observed at 3.56 ppm. According to the integral results, the molar ratio of 1,4-dioxane was 0.06 eq.

[0309] The XRPD results are shown in Figure 28. The compound of formula A-IV was in a crystal form, which was recorded as malonic acid salt type A. The specific XRPD analysis spectrum is shown in Table 22. The TGA results showed that the sample had a weight loss of about 0.49 wt% during heating to 130 °C (Figure 30). The DSC results showed that the sample had an endothermic peak with an onset temperature of 172.87 °C and a peak temperature of 176.14 °C (Figure 29).

[0310] Table 22

[0311] Example 39

[0312] The compound of formula (I) 40 mg was added with 0.6 ml 1,4-dioxane, and 13.4 mg malonic acid. After stirring at 50 °C for 3 hours, the temperature was decreased to room temperature, and the stirring was continued overnight. The solid was centrifuged and dried in a vacuum oven at 50 °C to obtain the compound of formula A-IV.

[0313] The NMR results showed that, compared with the free state, multiple signal peaks from 4.5 ppm to 5.5 ppm were shifted, indicating that the sample was salted. The NMR integral results were basically consistent with the raw material. The signal peak of malonic acid was observed near 3.16 ppm. According to the integral calculation, the salted ratio was 1:1.1 (i.e. n was 1.1). The solvent peak of 1,4-dioxane was observed at 3.56 ppm. According to the integral results, the molar ratio of 1,4-dioxane was 0.06 eq.

[0314] XRPD results are shown in Figure 31, the compound of formula A-IV is in a crystalline form, which is designated as malonate type B, and the XRPD analysis is shown in Table 23. TGA results show that the sample has a weight loss of about 0.19 wt% during heating to 130°C (Figure 33). DSC results show that the sample has an endothermic peak with an onset temperature of 168.00°C and a peak temperature of 171.90°C (Figure 32).

[0315] Table 23

[0316] Example 40

[0317] The compound of formula (I) 40 mg was added to 0.2 ml of ethanol, and 24.3 mg of p-toluenesulfonic acid monohydrate was added. After stirring at 50°C for 3 hours, it was allowed to cool to room temperature and stirred overnight. Then 0.6 ml of n-heptane was added, and the stirring was continued at room temperature for a certain period of time. After centrifugation, the solid was dried in a vacuum oven at 50°C to obtain the product.

[0318] The XRPD test results of the product (Figure 34) showed that it was in a crystalline form, and the XRPD analysis is shown in Table 24. This crystalline form is designated as p-toluenesulfonic acid salt type A of the compound of formula (I).

[0319] Table 24

[0320] The properties of the salt forms obtained in the above examples are summarized in Table 25.

[0321] Table 25 Summary of salt form properties

[0322] Test Example

[0323] The control compound in the test example is a compound known in the prior art, and its structure is:

[0324] Test Example 1

[0325] IC of NLRP3 inhibitors in reducing the level of IL-1β secreted by THP-1 cells 50 Experiment

[0326] 1. Experimental principle: This experiment uses human monocyte cell line THP-1 to study the inhibitory activity of NLRP3 inhibitors on the secretion of IL-1β by cells. Tocopheryl-Palmitate-12-myristate-13-acetate (PMA) is used to treat human THP-1 cells to differentiate into mature macrophage model. Then, according to the two stages of inflammasome formation in cells (assembly and activation), the cells are sequentially stimulated with lipopolysaccharide LPS and nigericin, which ultimately promotes the release of IL-1β from THP-1 cells. The first stage of inflammasome formation is to stimulate the cells with LPS (lipopolysaccharide), an agonist of Toll-like receptor TLR4, to activate the protein expression of inflammasome-related proteins NLRP3, caspase 1 and IL-1β precursor pro-IL-1β. The second stage of inflammasome formation is to add potassium ion carrier nigericin to change the membrane potential in THP-1 cells, destroy the membrane structure of the cells, cause the cells to expel potassium ions, further stimulate NLRP3 monomers to oligomerize to form NLRP3 oligomers, and begin to recruit ASC and pro-caspase-1 to assemble NLRP3 inflammasome complex containing a large amount of pro-caspase-1. Activated NLRP3 inflammasome activates pro-caspase-1 by proteolysis to form a large amount of active caspase-1, which further proteolyzes to promote the conversion of pro-IL-1β to mature IL-1β that can be secreted. In this process, NLRP3 inhibitors can effectively inhibit the maturation and activation of NLRP3 induced by nigericin, as well as the activation of caspase-1 downstream, thereby inhibiting the maturation and secretion of IL-1β.

[0327] 2. Experimental reagents and instruments

[0328] Table 26

[0329] 3. Experimental steps

[0330] (1) Collect cell suspension. Centrifuge the cells at 1000 rpm for 4 minutes, discard the supernatant, resuspend the cells with 1 mL of test medium, then gently blow the cell medium up and down with a pipette and a gun head to break the cell mass. After cell counting, resuspend THP-1 cells with complete RPMI medium 1640 containing PMA and inoculate into 96-well plates, and incubate in a 37°C, 5% CO2 incubator for 72 hours.

[0331] (2) Use an automatic plate washing dispenser EL406 to remove the medium from each well, and rinse each well twice with PBS. Add 96 μL of RPMI 1640 medium containing LPS to each well. Incubate the cells in a 37°C, 5% CO2 incubator for 4 hours.

[0332] (3) Add 2 μL of test compound and 2 μL of nigericin sodium salt to each well, and the final concentration of the compound screening is 5000 nM, 1000 nM, 200 nM, 40 nM, 8 nM, 1.6 nM, 0.32 nM, 0 nM, respectively. The cells are cultured in a 37°C, 5% CO2 incubator for 1 hour.

[0333] (4) Take an appropriate amount of supernatant, dilute it 100 times, and use the Human IL-1beta Valukine ELISA kit to detect the content of IL-1β in the supernatant.

[0334] 4. Experimental results

[0335] The experimental results are shown in Table 27.

[0336] Table 27 IC of NLRP3 inhibitor detected by THP-1 cells 50 Test results of the experiment

[0337] Conclusion: The compound represented by formula (I) has significant inhibitory activity on the maturation and secretion of IL-1β of THP-1 cells.

[0338] Test Example 2: Evaluation experiment of mouse acute peritonitis model

[0339] Male C57BL / 6 mice are orally administered with a dose of 10 mg / kg of the test compound or the control solvent, and then 1 hour later, 1 μg of E. coli 055: B5 ultrapure lipopolysaccharide (LPS) suspension is injected intraperitoneally, followed by 2 hours of intraperitoneal injection of 30 mM disodium salt ATP suspension 0.5 mL (in PBS, the pH of the ATP solution is adjusted to 7.2 before injection). After 30 minutes, all animals will be euthanized by carbon dioxide, and blood and peritoneal lavage fluid will be collected. The peritoneal lavage fluid is placed at 4°C.

[0340] Preparation of peritoneal lavage fluid: The peritoneal cavity is lavaged with ice-cold PBS (3 mL) containing 25 U / mL heparin and 10% heat-inactivated FBS. Before use, add one tablet of protease inhibitor cocktail to every 50 mL of buffer. The collected lavage fluid will be centrifuged (centrifugation conditions: 1000 g, 4°C, 10 minutes) to remove cells and cell debris, and the clarified sample will be stored at -80°C for cytokine IL-1β measurement.

[0341] The change of IL-1β cytokine level in the peritoneal lavage fluid and serum was determined using mouse IL-1β / IL-1F2 DuoSet ELISA kit (R&D Systems), and the inhibition rate of the compound on IL-1β was calculated by comparison with the Vehical group. The experimental results are shown in Table 28.

[0342] Table 28

[0343] Conclusion: The compound represented by formula (I) showed inhibitory effect on IL-1β in the animal in vivo model evaluation experiment.

[0344] Test Example 3 Evaluation test of MSU-induced gouty arthritis model in rats

[0345] In this test, male SD rats were used, and acute gouty arthritis model was established by intra-articular injection of 64 mg / mL sodium urate (MSU) crystal suspension into the right ankle joint of rats. The animals were randomly divided into normal control group, model control group, and different dose groups of test compound, 7 rats per group. The compound groups were orally administered 1 hour before and after modeling, and the normal control group and model control group were given the same amount of solvent. The foot circumference and foot diameter were measured before modeling and 24 h after modeling.

[0346] The experimental results are shown in Figure 1, and the compound represented by formula (I) showed dose-dependent improvement in the degree of ankle joint swelling in the MSU-induced acute gouty arthritis model in rats, and the effect was significantly better than that of the control compound.

[0347] Test Example 4 Solubility test

[0348] The following compounds were weighed and added to an appropriate amount of water, stirred at 37°C for 2 h, sampled, and the sampled solution was filtered with a 0.22 μm water filter membrane. For some samples with higher concentration, diluent was used for appropriate dilution. The signal peak area of the solution was measured by HPLC, and finally the concentration of the compound in the solution was calculated according to the peak area, HPLC standard curve of the raw material, and dilution factor. The results are shown in Table 29.

[0349] Table 29 Solubility results

[0350] In this test example, the HPLC test conditions are shown in Table 30:

[0351] Table 30

[0352] Test Example 5 Hygroscopicity evaluation

[0353] The hygroscopicity of the maleate salt type A, malonate salt type A, and malonate salt type B of the compound of formula (I) was evaluated by the following method, and the results are shown in Figures 35-37 (Table 31).

[0354] Dynamic water sorption-desorption analysis was determined by DVS Intrinsic (SMS, UK). The test was performed in gradient mode with a humidity change of 0%-95%-0%, and the humidity change was 10% in the range of 0% to 90% for each gradient. The end of the gradient was determined by dm / dt, and the end of the gradient was determined when dm / dt was less than 0.002% and maintained for 10 minutes.

[0355] Table 31

[0356] Test Example 6 Dynamic solubility test in biological media and water

[0357] The preparation process of the biological medium is shown in Table 32. The samples of different crystal forms were added to the biological medium and water and stirred at 37°C for 24h. Samples were taken at 0.5h, 2h and 24h, respectively. The sampled solution was filtered with a 0.22μm water filter membrane, and some samples with high concentration were appropriately diluted with diluent. The signal peak area of the solution was measured by HPLC, and finally the concentration of the compound in the solution was calculated according to the peak area, the HPLC standard curve of the raw material and the dilution factor. In addition, the 24h supernatant was tested for pH value. The test results are shown in Table 33.

[0358] Table 32 Preparation process of biological medium

[0359] Table 33 Solubility results in biological medium and water

[0360] Test Example 7 Stability study

[0361] The crystal form samples were placed in high temperature (60°C), high humidity (25°C / 92.5% RH), and light (25°C / 4500Lux) conditions for stability study. Samples were taken at 7th day and 14th day for XRPD characterization and HPLC test, and the results are shown in Table 34.

[0362] Table 34

[0363] Test Example 8 Pharmacodynamic test in DIO obese mouse model induced by high-fat diet

[0364] 1. Purpose of the experiment

[0365] The pharmacodynamic effect of the test compound on the DIO mouse model induced by high-fat diet was tested.

[0366] 2. Test materials

[0367] 2.1 Reagents

[0368] Hematoxylin staining solution: Zhongshan Golden Bridge Biotechnology Co., Ltd., Catalog No.: ZLI-9610.

[0369] Water-soluble eosin staining solution: Zhongshan Golden Bridge Biotechnology Co., Ltd., Catalog No.: ZLI-9613.

[0370] Sirius red staining solution: Beijing Solerbio Science and Technology Co., Ltd., Catalog No.: G1427.

[0371] 2.2 Equipment

[0372] Electronic balance: Sartorius, Model: Quintix 124-1CN

[0373] Electronic scale: Shanghai Yueping Scientific Instrument Co., Ltd., Model: YP10001

[0374] Ultrasonic cleaner: Kunshan Ultrasonic Instrument Co., Ltd., Model: KQ3200E

[0375] Shaker: Itslin, Model: 01-00X-1012

[0376] High-speed homogenizer: IKA, Model: T10 basic

[0377] Anesthesia machine: MIP, Model: 50318

[0378] Refrigerated centrifuge: Thermo Fisher, Catalog No.: Sorvall Legend Micro 17r

[0379] Multifunctional enzyme marker: TECAN, Catalog No.: SPARK 10M

[0380] Microscope: OLYMPUS, Catalog No.: DP720

[0381] Tissue dehydrator: Leica, Catalog No.: TP1020

[0382] Paraffin embedding machine: Leica, Catalog No.: EG1150H

[0383] Freeze table: Leica, Catalog No.: EG1150C

[0384] Microtome: Leica, Catalog No.: RM2235

[0385] 3. Experimental methods

[0386] 3.1 Experimental grouping

[0387] Table 35. Animal grouping and dosing schedule

[0388] 3.2 Model establishment and drug treatment

[0389] After the animals arrived and adapted to the environment for 1 week, the animals were randomly divided into 2 groups (7+28), 7 mice were normally fed, and 28 mice were fed with high-fat feed (HFD). After the animals were raised for 22-23 weeks, they were fasted for 6 hours, and the blood glucose level was detected by blood glucose meter. All animals were subjected to orbital blood collection, and serum was collected by centrifugation to detect serum TC and TG levels. The animals were randomly divided into 4 groups according to body weight, serum TC and TG levels, and continued to be fed with high-fat feed (HFD), and another group of 7 mice were normally fed. According to Table 4, the drug was administered for 30 days.

[0390] 3.3 Reagent preparation

[0391] 1) Semaglutide stock solution (0.5 mg / ml): Semaglutide 1.0 mg was weighed, 2 ml PBS was added, and it was slowly blown and beaten to dissolve to prepare a stock solution with a concentration of 0.5 mg / ml.

[0392] ■Semaglutide low dose (0.0125 mg / ml): 25 uL of stock solution was pipetted, 975 uL of PBS was added, and a liquid with a concentration of 0.0125 mg / ml was prepared for immediate use.

[0393] 2) 0.5% carboxymethylcellulose sodium (CMC-Na) solution preparation (for dissolution of test sample GS8-164): 0.5 g of CMC-Na powder was slowly added to 100 mL of ultrapure water, and was magnetically stirred and dissolved with heating, and was mixed evenly. After filtration with a 0.22 mm filter membrane, it was stored at 2-8°C for later use.

[0394] ■Compound of formula (I) (2 mg / mL, content: 96.7%): 84.0 mg of compound of formula (I) was weighed, 40.614 mL of 0.5% CMC-Na was added, a magnetic stirrer was added to the magnetic stirrer for sufficient stirring, and was vortexed and ultrasonicated for 20 minutes to make the drug preparation into a suspension state. It was prepared once every two days, and was stored at 2-8°C for later use.

[0395] 3.4 Detection index

[0396] Continuous administration for 1 month, body weight was measured every 3 days during the administration period, and the experimental results are shown in Figure 38.

[0397] The compound of formula (I) at a dose of 20mpk reduced weight by 5% in 30 days compared to the model group. The weight loss effect (33%) of the compound of formula (I) (20mpk) combined with Semaglutide (0.025mpk) was better than that of Semaglutide (0.025mpk) alone (23%).

[0398] Conclusion: The compound of formula (I) showed good weight loss effect in the DIO obesity model, and after being combined with Semaglutide, the weight loss effect was better than that of Semaglutide alone, showing excellent potential for combination therapy.

[0399] The above describes the embodiments of the present application. However, the present application is not limited to the above-described embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A compound having the structure shown in Formula A: in, X represents acid, and n is a number from 0.1 to 4.

2. The compound according to claim 1, characterized in that, The acid is an inorganic acid or an organic acid. For example, the inorganic acid is selected from one or more of hydrochloric acid, sulfuric acid, phosphoric acid, and hydrobromic acid, and the organic acid is selected from one or more of methanesulfonic acid, lactic acid, maleic acid, malic acid (e.g., L-malic acid), tartaric acid (e.g., L-tartaric acid), p-toluenesulfonic acid, fumaric acid, ethanesulfonic acid, malonic acid, and oxalic acid.

3. The compound according to claim 1 or 2, characterized in that, The n is a number between 0.4 and 3.

4. The compound according to claim 1, characterized in that, When X represents hydrochloric acid, n is a number between 0.7 and 1.3; When X represents sulfuric acid, n is a number between 0.7 and 1.3; When X represents phosphoric acid, n is a number ranging from 0.5 to 2.5; When X represents methanesulfonic acid, n is a number between 0.7 and 1.3; When X represents hydrobromic acid, n is a number ranging from 0.5 to 2.5; When X represents maleic acid, n is a number between 0.9 and 1.3; When X represents fumaric acid, n is a number ranging from 0.4 to 1.3; When X represents L-malic acid, n is a number ranging from 0.5 to 1.3; When X represents L-tartaric acid, n is a number ranging from 0.5 to 1.3; When X represents malonic acid, n is a number ranging from 0.5 to 1.3; When X represents p-toluenesulfonic acid, n is a number between 0.5 and 1.3; For example, the compound is a compound of formula A1, a compound of formula A-II, a compound of formula A-III, or a compound of formula A-IV; The AI ​​compound has the following structure: n is selected from a number between 0.6 and 1.8, preferably a number between 0.9 and 1.3, and more preferably a number between 0.9 and 1; The compound of formula A-II has the following structure: n is selected from a number between 0.6 and 1.8, preferably a number between 0.9 and 1.3, and more preferably a number between 0.9 and 1.1; The compound of formula A-III has the following structure: n is selected from a number between 0.5 and 1.5, preferably a number between 0.9 and 1.3; The compound of formula A-IV has the following structure: n is selected from a number between 0.5 and 1.5, preferably a number between 0.9 and 1.3, and more preferably a number between 1.04 and 1.

1.

5. A method for preparing the compound according to any one of claims 1-4, characterized in that, include: The compound shown in formula (I) is reacted with an appropriate amount of acid X to obtain the compound shown in formula A; 6. The preparation method according to claim 5, characterized in that, The reaction is carried out in an organic solvent, for example, the organic solvent is selected from one or more of ethanol, acetone, 1,4-dioxane, and n-heptane; And / or, the temperature of the reaction is 40–60°C.

7. The crystal form of the compound of formula AI, characterized in that, Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 8.1±0.20°, 8.4±0.20°, 13.4±0.20°, 21.0±0.20°, and 22.7±0.20°; further, characteristic peaks are also observed at 5.4±0.20°, 6.9±0.20°, 9.1±0.20°, 13.9±0.20°, 18.0±0.20°, 18.9±0.20°, and / or 24.6±0.20°; even further, characteristic peaks are observed at 11.0±0.20°, 11.2±0.20°, Characteristic peaks are present at 14.5±0.20°, 16.6±0.20°, 17.4±0.20°, 18.4±0.20°, 20.0±0.20°, 21.9±0.20°, 23.2±0.20°, 23.5±0.20°, 25.5±0.20°, 25.9±0.20°, 26.2±0.20°, 26.6±0.20°, 27.0±0.20°, 27.7±0.20°, 28.0±0.20°, 29.4±0.20°, 30.1±0.20° and / or 30.5±0.20°. The AI ​​compound has the following structure: n is a number selected from 0.6 to 1.8, preferably a number from 0.9 to 1.3, and for example a number selected from 0.9 to 1; Preferably, the crystal form of the AI ​​compound has an XRPD pattern as shown in Figure 13; Preferably, the crystal form of the AI ​​compound is anhydrous.

8. The crystal form of the compound of formula A-II, characterized in that, Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 8.2±0.20°, 8.7±0.20°, 13.7±0.20°, 19.4±0.20°, 21.3±0.20°, and 23.1±0.20°; further, characteristic peaks are also observed at 5.5±0.20°, 7.0±0.20°, 9.4±0.20°, 18.2±0.20°, and / or 25.1±0.20°; even further, characteristic peaks are observed at 10.8±0.20°, 11.3±0.20°, and 14.1±0.20°. Characteristic peaks are present at 0.20°, 16.9±0.20°, 18.9±0.20°, 19.8±0.20°, 20.6±0.20°, 22.1±0.20°, 22.4±0.20°, 22.7±0.20°, 25.7±0.20°, 26.2±0.20°, 27.8±0.20°, 28.3±0.20°, 30.0±0.20°, 30.7±0.20°, 31.6±0.20°, 32.3±0.20°, 32.5±0.20° and / or 32.7±0.20°. The compound of formula A-II has the following structure: n is a number selected from 0.6 to 1.8, preferably a number from 0.9 to 1.3, and for example a number selected from 0.9 to 1.1 (e.g., 1); Preferably, the crystal form of the compound of formula A-II has an XRPD pattern as shown in Figure 19; Preferably, the compound of formula A-II is anhydrous.

9. The crystal form of the compound of formula A-III, characterized in that, Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 8.3±0.20°, 9.0±0.20°, 13.9±0.20°, 21.3±0.20°, 23.1±0.20°, and 23.7±0.20°; further, characteristic peaks are also observed at 9.6±0.20°, 11.5±0.20°, 14.8±0.20°, 18.3±0.20°, and / or 19.6±0.20°. The compound of formula A-III has the following structure: n is selected from a number between 0.5 and 1.5, preferably a number between 0.9 and 1.3, and is 1 for example; Preferably, the crystal form of the compound of formula A-III has an XRPD pattern as shown in Figure 28; Preferably, the crystal form of the compound of formula A-III is anhydrous.

10. The crystal form of the compound of formula A-IV, characterized in that, Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 8.7±0.20°, 14.5±0.20°, 19.0±0.20°, 22.1±0.20°, 22.7±0.20°, and 24.0±0.20°; further, characteristic peaks are also observed at 7.3±0.20°, 9.5±0.20°, 11.9±0.20°, 20.5±0.20°, 21.1±0.20°, 23.3±0.20°, and / or 31.2±0.20°; even further, characteristic peaks are also observed at 10.0±0.20°, 26.2±0.20°, 29.3±0.20°, and / or 29.7±0.20°. The compound of formula A-IV has the following structure: n is selected from a number between 0.5 and 1.5, preferably a number between 0.9 and 1.3, and for example a number between 1.04 and 1.1; Preferably, the crystal form of the compound of formula A-IV has an XRPD pattern as shown in Figure 31; Preferably, the crystal form of the compound of formula A-IV is anhydrous.

11. A pharmaceutical composition, characterized in that, It comprises one or more of the following: the compound of any one of claims 1-4, the crystal form of the compound of formula AI of claim 7, the crystal form of the compound of formula A-II of claim 8, the crystal form of the compound of formula A-III of claim 9, and the crystal form of the compound of formula A-IV of claim 10.

12. The pharmaceutical composition according to claim 11, characterized in that, The pharmaceutical composition further comprises one, two or more pharmaceutically acceptable excipients; and / or, the pharmaceutical composition further comprises additional therapeutic agents for use in combination with the compound shown in Formula A; Preferably, the additional therapeutic agent is selected from smegglutinin.

13. Use of the compound of any one of claims 1-4, the crystal form of the compound of formula AI of claim 7, the crystal form of the compound of formula A-II of claim 8, the crystal form of the compound of formula A-III of claim 9, the crystal form of the compound of formula A-IV of claim 10, or the pharmaceutical composition of any one of claims 11-12 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 one or more of the following: a compound of formula A, a crystal form of a compound of formula A1, a crystal form of a compound of formula A-II, a crystal form of a compound of formula A-III, and a crystal form of a compound of formula A-IV, as well as smegglutinin.

14. The use according to claim 13, 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).

15. A method for treating obesity, characterized in that, The method includes administering to a patient a preventive or therapeutically effective amount of the compound of any one of claims 1-4, the crystal form of the compound of formula AI of claim 7, the crystal form of the compound of formula A-II of claim 8, the crystal form of the compound of formula A-III of claim 9, the crystal form of the compound of formula A-IV of claim 10, or the pharmaceutical composition of any one of claims 11-12. Preferably, the pharmaceutical composition comprises one or more of the following: a compound of formula A, a crystal form of a compound of formula A1, a crystal form of a compound of formula A-II, a crystal form of a compound of formula A-III, and a crystal form of a compound of formula A-IV, as well as smegglutinin.

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