Sulfonylurea derivative having NLRP3 inhibitory activity and use thereof

Abstract

Disclosed in the present invention are a compound as shown in formula 1, or a tautomer, mesomer, racemate, enantiomer and diastereomer thereof, a mixture form thereof, or a pharmaceutically acceptable salt thereof. The compound has excellent NLRP3 inhibitory activity, and the compound of the present invention lays a new material basis for the development of therapeutic drugs for NLRP3-related diseases.

Description

Sulfonylurea derivatives with NLRP3 inhibitory activity and applications thereof Technical Field

[0001] The present invention relates to the field of medicinal chemistry. Specifically, the present invention relates to sulfonylurea derivatives having NLRP3 inhibitory activity, a drug combination comprising the sulfonylurea derivatives, and their use in gout medicine. Background Art

[0002] The NLRP3 inflammasome is a complex with a molecular weight of approximately 700,000 composed of the nucleotide-binding oligomerization domain (NOD)-like receptor family, pyrin domain-containing protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and caspase-1. The NLRP3 inflammasome is a multimeric protein complex primarily expressed in monocytes / macrophages, neutrophils, and dendritic cells. Structurally, the NLRP3 sensor protein comprises three domains: a leucine-rich repeat receptor (LRR) domain, a nucleotide-binding and oligomerization (NACHT) domain, and a pyrin domain (PYD). The LRR at the carboxyl terminus is responsible for detecting and monitoring microbial ligands and endogenous signaling molecules. The NACHT domain is crucial for oligomerization of the NLRP3 inflammasome and functions as an ATPase. The Walker A motif contains an ATP-binding site, and the Walker B motif is essential for ATPase activity. Small molecules with Michael receptor moieties typically form covalent bonds with cysteine ​​residues of the ATPase, resulting in irreversible inhibition. Therefore, NLRP3 inhibitors primarily target the ATPase. The amino-terminal PYD domain can homotypically interact with ASC and participate in inflammasome assembly.

[0003] Activation of the NLRP3 inflammasome involves two steps: priming and activation. During the priming phase, bacterial endotoxins such as lipopolysaccharide activate the TLR4 receptor, which, through the nuclear factor-κB pathway, upregulates the gene expression of NLRP3 and inflammatory cytokine precursors such as pro-IL-1β and pro-IL-18. During the activation phase, when the LRR domain recognizes the "danger" signal, the amino-terminal PYD interacts with the PYD domain of ASC. This interaction then recruits pro-caspase-1, cleaving pro-caspase-1 into active caspase-1, thereby converting pro-IL-1β and pro-IL-18 into their biologically active forms, triggering inflammation. Caspase-1 also cleaves gasdermin D (GSDMD) into N-terminal gasdermin-D, creating membrane pores and inducing pyroptosis. NIMA-related kinase NEK7 acts downstream of P2X7 and potassium efflux, binding to the LRR domain of NLRP3 through its catalytic domain to form the NLRP3-NEK7 complex, regulating the assembly and activation of the NLRP3 inflammasome.

[0004] The specific mechanisms of NLRP3 inflammasome activation remain unclear, but several proposed models include potassium ion efflux, chloride ion efflux, calcium ion signaling, lysosomal rupture, ROS production, and mitochondrial dysfunction. Abnormal NLRP3 inflammasome activation has been linked to the development of numerous diseases, such as various metabolic syndromes and metabolic disorders, including gout, non-alcoholic steatohepatitis, atherosclerosis, and type 2 diabetes. It is also implicated in the development and progression of neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, and multiple sclerosis. Furthermore, it plays a role in the formation of immune-inflammatory disorders such as inflammatory bowel disease and the genetic disease cryoproliferative syndrome, caused by NLRP3 gene mutations.

[0005] Although several inhibitors targeting NLRP3 inflammasome activation are currently available, they are often due to unclear targets, significant side effects, and poor drugability. For example, the NLRP3 inhibitor MCC950 was withdrawn from clinical trials due to hepatotoxicity. Therefore, the development of new NLRP3 inflammasome inhibitors is urgently needed to provide better options for treating diseases involving the NLRP3 inflammasome. Currently, there are no drugs marketed that inhibit the NLRP3 pathway to treat gout and other diseases related to this pathway, such as non-alcoholic steatohepatitis, type 2 diabetes, and inflammatory bowel disease. Therefore, the development of new NLRP3 inhibitor compounds is of great significance for the treatment of these diseases, especially gout. Summary of the Invention

[0006] The object of the present invention is to provide a compound having NLRP3 inhibitory activity.

[0007] Another object of the present invention is to provide a pharmaceutical composition comprising the compound.

[0008] Another object of the present invention is to provide a use of the compound in the preparation of a drug for treating NLRP3-related diseases and a method for treating NLRP3-related diseases using the compound or pharmaceutical composition.

[0009] In a first aspect, the present invention provides a compound of Formula 1, or a tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt thereof,

[0010] Where,

[0011] R1 is selected from: H, halogen, cyano, hydroxy, substituted or unsubstituted C 1-3 Amide, substituted or unsubstituted sulfonamide, substituted or unsubstituted C 1-3 Acyl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted C 5-10 aryl;

[0012] R2 is selected from substituted or unsubstituted C 5-16 Aryl or heteroaryl or cycloalkyl or heterocycloalkyl.

[0013] In a preferred embodiment, the heteroaryl or heterocycloalkyl group has 1, 2 or 3 heteroatoms independently selected from O, N or S in the ring.

[0014] In a specific embodiment, R2 is selected from the group consisting of substituted or unsubstituted pyrazolyl, substituted or unsubstituted furanyl, substituted or unsubstituted tetrahydrofuranyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted pyranyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted triazolyl, substituted or unsubstituted tetrazolyl, substituted or unsubstituted imidazolyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted morpholinyl, substituted or unsubstituted piperazinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted phenyl, substituted or unsubstituted phenylheteroaryl, substituted or unsubstituted phenylheterocyclyl, substituted or unsubstituted biphenyl, substituted or unsubstituted quinolyl, substituted or unsubstituted isoquinolyl, substituted or unsubstituted naphthyl, substituted or unsubstituted pyrazinyl, and substituted or unsubstituted pyrimidinyl.

[0015] In a specific embodiment, R2 is selected from: substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted quinolyl, substituted or unsubstituted quinoxalinyl, substituted or unsubstituted phenylheteroaryl, substituted or unsubstituted phenylheterocyclyl, substituted or unsubstituted biphenyl, substituted or unsubstituted thienyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted pyrrolidinyl.

[0016] In a specific embodiment,

[0017] R2 is

[0018] R3 and R 31 independently selected from: H, hydroxy, halogen, cyano, nitro, substituted or unsubstituted 5-7 membered heterocyclic ring containing 1-2 heteroatoms independently selected from N, O or S, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 5-10 Aryl or heteroaryl containing 1-2 heteroatoms independently selected from N, O or S, a sulfonamide group containing 1-2 heteroatoms independently selected from N, O or S;

[0019] Alternatively, two adjacent R3 together with the carbon atoms to which they are attached form a 3-10 membered saturated ring, a substituted or unsubstituted C 5-10 Aromatic ring (e.g. benzene ring) or 3-10 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S or C 5-10 aromatic heterocycles;

[0020] x = 1, 2, 3, 4, or 5;

[0021] y = 1, 2, 3, or 4;

[0022] R4 is selected from the group consisting of: H, OH, substituted or unsubstituted amino, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted C 5-10 Aryl, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

[0023] R5 is selected from: H, substituted or unsubstituted C 1-3 Acyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 5-10 Aryl, substituted or unsubstituted C 3-10Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

[0024] R6 and R7 are independently selected from: H, substituted or unsubstituted C 1-3 Acyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 5-10 Aryl, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

[0025] n and m are independently selected from 0, 1, 2, 3, 4, 5 or 6.

[0026] In a specific embodiment,

[0027] R1 is selected from: H, halogen, cyano;

[0028] R2 is

[0029] R3 is selected from: halogen, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-4 Alkoxy, substituted or unsubstituted C 5-10 Aryl (e.g. phenyl), -substituted or unsubstituted C 1-4 Alkyl-OH, -N(R 61 )-substituted or unsubstituted C 1-6 Alkyl-NR 62 R 72 , -O-substituted or unsubstituted C 1-6 Alkyl-NR 62 R 72 ,

[0030] Alternatively, two adjacent R3 together with the carbon atom to which they are attached form a substituted or unsubstituted C 5-10 an aromatic ring (e.g., a benzene ring) or a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

[0031] x = 1, 2, or 3;

[0032] R 61 Selected from: H, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-3 acyl group;

[0033] R 62 and R 72 Independently selected from: H, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C1-3 Acyl, or R 62 and R 72 Together with the nitrogen atom to which they are attached, they form a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S.

[0034] In a specific embodiment,

[0035] R1 is selected from: H;

[0036] R2 is

[0037] R3 is selected from: halogen (such as Br), nitro, substituted or unsubstituted C 1-6 Alkyl, -N(R 61 )-substituted or unsubstituted C 1-6 Alkyl-NR 62 R 72 ,

[0038] Alternatively, two adjacent R3 together with the carbon atom to which they are attached form a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O or S;

[0039] x = 1, 2, or 3;

[0040] R 61 Selected from: H, substituted or unsubstituted C 1-4 alkyl;

[0041] R 62 and R 72 Independently selected from substituted or unsubstituted C 1-6 Alkyl; or, R 62 and R 72 Together with the nitrogen atom to which they are attached, they form a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S.

[0042] In a specific embodiment, the compound, or a tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0043] In a specific embodiment, the compound is selected from the group consisting of:

[0044] Preferably, the compound is selected from the group consisting of:

[0045] In a second aspect, the present invention provides a pharmaceutical composition, characterized in that the pharmaceutical composition comprises the compound described in the first aspect or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, and optionally a pharmaceutically acceptable carrier.

[0046] In a third aspect, the present invention provides use of the compound according to the first aspect or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt in the preparation of an NLRP3 inhibitor.

[0047] In a specific embodiment, the NLRP3 inhibitor is a drug for treating and / or preventing NLRP3-related diseases.

[0048] In a specific embodiment, the NLRP3-related diseases include but are not limited to gout, neurodegenerative diseases, non-alcoholic steatohepatitis, type II diabetes, inflammatory bowel disease, hepatitis, and arthritis.

[0049] In a preferred embodiment, the neurodegenerative disease includes but is not limited to Alzheimer's disease and Parkinson's disease.

[0050] In a fourth aspect, the present invention provides a method for treating an NLRP3-related disease, comprising the step of administering a therapeutically effective amount of the compound of the first aspect or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, or the pharmaceutical composition of the second aspect to a subject in need thereof.

[0051] In a preferred embodiment, the NLRP3-related diseases include but are not limited to gout, neurodegenerative diseases, non-alcoholic steatohepatitis, type II diabetes, inflammatory bowel disease, hepatitis, and arthritis.

[0052] In a preferred embodiment, the neurodegenerative disease includes but is not limited to Alzheimer's disease and Parkinson's disease.

[0053] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] [Corrected 14.05.2025 according to Rule 91] Figures 1A-1C Compound N21 Na + Effects on relevant validation indicators of the MSU-induced mouse foot gout model;

[0055] Figure 2 Compound N21 Na + and MCC950 on the liver and kidney functions of mice. DETAILED DESCRIPTION

[0056] After extensive and in-depth research, the inventors unexpectedly discovered a series of compounds with NLRP3 inhibitory activity, which can be used to prepare NLRP3-related disease therapeutic drugs, thereby treating NLRP3-related diseases. The compounds of the present invention have excellent NLRP3 inhibitory activity, and the inhibitory activity of some compounds is significantly better than that of the positive control, and the IC of NLRP3 inhibition is 50 The value can even reach nm level. On this basis, the present invention was completed.

[0057] Definition of terms

[0058] The terms used herein with respect to the groups, substituents or structures of the compounds have the same meanings as understood by those skilled in the art. For the sake of clarity, the terms used in this specification are defined as follows.

[0059] As used herein, “a” or “an” or “a type” means at least one / kind / type or one / kind / type or more than one / kind / type.

[0060] In this article, the form "C 1-n " means that the group has 1-n carbon atoms, for example, "C 1-10 " means that the group has 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms; similarly, "C6-C10" means that the group has 6, 7, 8, 9 or 10 carbon atoms. At the same time, the description of the range of carbon atoms herein also includes the sub-ranges therein. For example, when 1-10 carbon atoms are mentioned herein, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, and 1-3 carbon atoms are also included.

[0061] The term "alkyl" as used herein has the same meaning as commonly understood by those of ordinary skill in the art, and refers to various saturated or unsaturated straight-chain, side-chain, or cyclic hydrocarbon groups. For example, the alkyl group described herein refers to a lower alkyl group having 1 to 10 carbon atoms; preferably a lower alkyl group having 1 to 8 carbon atoms; more preferably a lower alkyl group having 1 to 6 carbon atoms. In specific embodiments, the alkyl group described herein includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, and the like. Similarly, the terms "alkenyl" or "alkynyl" as used herein refer to various unsaturated straight-chain, side-chain, or cyclic hydrocarbon groups containing a carbon-carbon double bond or a carbon-carbon triple bond.

[0062] As used herein, the terms "aryl" and "aromatic ring" have the same meaning as commonly understood by those skilled in the art, and refer to a cyclic conjugated aromatic system. For example, the term "C6-C10 aryl" refers to an aromatic ring group having 6 to 10 carbon atoms, such as phenyl and naphthyl, that does not contain heteroatoms in the ring. The term "heteroaryl" as used herein refers to a cyclic conjugated aromatic system that contains one or more heteroatoms, such as N, O, or S, in the ring; for example, pyridyl and pyrazinyl.

[0063] The term "halogen" as used herein has the meaning commonly understood by those skilled in the art. In specific embodiments, halogen refers to fluorine, chlorine, bromine or iodine.

[0064] As used herein, the term "substituted" refers to the replacement of one or more hydrogen atoms on a specific group with a specific substituent. The specific substituent may be the substituent described above or the specific substituents described in the Examples. Therefore, in the present invention, the substituents in Formula 1 may each independently be the corresponding groups described in the specific compounds described in the Examples; that is, the present invention includes combinations of the substituents in Formula 1, as well as combinations of some of the substituents shown in Formula 1 with other specific substituents described in the Examples.

[0065] Unless otherwise specified, a substituted group may have a specific substituent at any substitutable position on the group, and the substituents may be the same or different at each position. A cyclic substituent, such as a heterocyclyl, may be attached to another ring, such as a cycloalkyl, to form a spirobicyclic ring system, e.g., where the two rings share a common carbon atom.

[0066] In particular, the various substituents defined above also include groups formed by further substitutions thereof, wherein these new substituents may also contain other groups. For example, hydrogen atoms on alkyl and aryl groups replaced by amino, halogen or other groups become groups within the above definitions.

[0067] In a specific embodiment, the "substituted or unsubstituted" or "optionally substituted" refers to being optionally substituted by one or more substituents selected from the following: cyano, halogen, hydroxyl, optionally substituted amino, nitro, carboxyl, ester, sulfonyl, oxo, deuterated, optionally substituted C 1-3 Alkyl, optionally substituted C 1-3 Alkoxy, optionally substituted C 1-3 Acyloxy, optionally substituted C 5-10 aryl, optionally substituted 3-7 membered cycloalkyl or heterocyclic group, optionally substituted sulfonylamino, optionally substituted acyl.

[0068] Compounds of the present invention

[0069] To overcome the deficiencies in the prior art, the present invention provides sulfonylurea derivatives, drug combinations thereof, and applications thereof that can be used for NLRP3-related diseases.

[0070] To this end, the present invention provides a sulfonylurea derivative, the structural formula of the sulfonylurea derivative is shown in Formula 1:

[0071] The substituents in the formula are each as described above.

[0072] Based on the compounds of the present invention, the present invention also provides a pharmaceutical composition comprising the above-mentioned compound or its tautomers, mesomers, racemates, enantiomers, diastereomers, mixtures thereof or pharmaceutically acceptable salts thereof, and optionally a pharmaceutically acceptable carrier.

[0073] Based on the teachings of the present invention, those skilled in the art will understand that the compounds of the present invention can be used to prepare NLRP3 inhibitors. In a specific embodiment, the NLRP3 inhibitors of the present invention are drugs for treating and / or preventing NLRP3-related diseases.

[0074] Specific NLRP3-associated diseases are known to those skilled in the art, including but not limited to gout, neurodegenerative diseases, nonalcoholic steatohepatitis, type II diabetes, inflammatory bowel disease, hepatitis, and arthritis. In preferred embodiments, the neurodegenerative diseases include but are not limited to Alzheimer's disease and Parkinson's disease. Of particular interest to the present invention are gout and other diseases.

[0075] Pharmaceutical compositions and treatment methods of the present invention

[0076] The compounds of the present invention possess excellent NLRP3 inhibitory activity and can be used as NLRP3 inhibitors, and further as drugs for treating the aforementioned NLRP3-related diseases. In view of this, in addition to the above-mentioned compounds, the present invention also provides a pharmaceutical composition comprising the compound of Formula I, or its tautomers, mesomers, racemates, enantiomers, diastereomers, mixtures, or pharmaceutically acceptable salts, and optionally a pharmaceutically acceptable excipient.

[0077] In a specific embodiment, the pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" refers to:

[0078] The amount of the compound is sufficient to significantly improve the condition without causing serious side effects.

[0079] "Pharmaceutically acceptable excipients or carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0080] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0081] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0082] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0083] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0084] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and flavoring agents.

[0085] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0086] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0087] Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0088] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds. When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage at the time of administration is a pharmaceutically effective dosage. The compounds and pharmaceutical compositions of the present invention can be administered by oral, nasal, skin, lung or gastrointestinal tract routes. Oral administration is most preferred, either in a single dose or in divided doses. Regardless of the method of administration, the optimal dose for an individual should be determined based on the specific treatment. Generally, the dose is started with a small dose and gradually increased until the most suitable dose is found. Of course, the specific dose should also take into account factors such as the route of administration and the patient's health status, which are all within the skill of a skilled physician.

[0089] Based on the teachings of the present invention, those skilled in the art can utilize the compounds or pharmaceutical compositions of the present invention to treat NLRP3-related diseases. In specific embodiments, the method comprises administering a therapeutically effective amount of a compound of Formula I, or a tautomer, mesomer, racemate, enantiomer, diastereomer, mixture, or pharmaceutically acceptable salt thereof, to a patient in need thereof. NLRP3-related diseases include, but are not limited to, gout, neurodegenerative diseases, nonalcoholic steatohepatitis, type II diabetes, inflammatory bowel disease, hepatitis, and arthritis; in particular, gout.

[0090] Advantages of the present invention:

[0091] 1. The compounds of the present invention have excellent NLRP3 inhibitory activity;

[0092] 2. The compounds of the present invention lay a new material foundation for the development of therapeutic drugs for NLRP3-related diseases.

[0093] 3. The compounds of the present invention can significantly alleviate gout symptoms and have good safety.

[0094] The technical solutions of the present invention are further described below with reference to specific examples. However, the following examples do not constitute a limitation of the present invention. All various application methods adopted in accordance with the principles and technical means of the present invention are within the scope of the present invention. Experimental methods in the following examples, where specific conditions are not specified, are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0095] All raw materials of the present invention are not particularly limited in their sources and can be purchased from the market or prepared according to conventional methods well known to those skilled in the art.

[0096] Example 1: Synthesis of Butylamine Sulfonylurea Compounds N01-N04

[0097] Phenbutylamine (1 eq) was dissolved in dichloromethane and stirred at 0°C for 10 min. Chlorosulfonic acid (4 eq) was added dropwise. The reaction was warmed to room temperature and monitored by TLC. After completion, the reaction was poured into ice-cold brine to quench. The mixture was then extracted three times with dichloromethane (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the sulfonyl chloride intermediate. The sulfonyl chloride compound (1 eq) was then dissolved in 1,4-dioxane. Ammonia in dioxane (5 eq) was added with stirring at room temperature. The reaction was allowed to react at room temperature for 2 h. After completion of the reaction by TLC, the reaction solution was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the sulfonamide intermediate.

[0098] Compound 1,2,3,5,6,7-hexahydro-s-indacene-4-amine (1 eq) was dissolved in an appropriate amount of THF, and TEA (1.1 eq) was added under stirring. Finally, triphosgene (0.3 eq) was added. The mixture was reacted in an oil bath at 75°C for 2 h. After completion of the reaction by TLC, the mixture was directly concentrated under reduced pressure and washed with n-pentane. The filtrate was collected and concentrated under reduced pressure to obtain intermediate 1.

[0099] The sulfonamide intermediate (1 eq) was then dissolved in THF, and NaH (1.2 eq) was added at 0°C. After 10 min, the isocyanate intermediate (1.2 eq) was added and reacted at room temperature for 2 h. After TLC detection, the reaction was completed and extracted with ethyl acetate three times (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the final product.

[0100] Example 2: Synthesis of Ethylamine Sulfonylurea Compound N05

[0101] 2-(Naphthyl-1-yl)ethylamine (1 eq) was dissolved in dichloromethane and stirred at 0°C for 10 min. Chlorosulfonic acid (4 eq) was added dropwise. The reaction was warmed to room temperature and monitored by TLC. After completion, the reaction was poured into ice-cold brine to quench. The mixture was then extracted three times with dichloromethane (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the sulfonyl chloride intermediate. The sulfonyl chloride compound (1 eq) was then dissolved in 1,4-dioxane. Ammonia in dioxane (5 eq) was added with stirring at room temperature. The reaction was allowed to react at room temperature for 2 h. After completion of the reaction by TLC, the reaction solution was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the sulfonamide intermediate. The sulfonamide intermediate (1 eq) was then dissolved in THF, and NaH (1.2 eq) was added at 0°C. After 10 min, the prepared isocyanate intermediate (1.2 eq) was added. The preparation method was the same as the synthesis operation of intermediate 1 above. The reaction was carried out at room temperature for 2 h. After TLC detection, the reaction was extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then purified by column chromatography to obtain the final product.

[0102] Example 3. Preparation of cyclic sulfonylurea compounds N06-N75, S1-S6

[0103] A sulfonyl chloride compound (1 eq) was dissolved in 1,4-dioxane. A solution of ammonia in dioxane (5 eq) was added with stirring at room temperature. The mixture was allowed to react at room temperature for 2 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the sulfonamide intermediate. The sulfonamide intermediate (1 eq) was then dissolved in THF. NaH (1.2 eq) was added at 0°C. After 10 min, intermediate 1 was added. The mixture was allowed to react at room temperature for 2 h. After completion of the reaction, the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the final product. Alternatively, if a solid precipitated after the reaction, the resulting solid was filtered and slurried / recrystallized with an appropriate solvent to obtain the final product.

[0104] Example 4: Preparation of sulfonylurea compound N76

[0105] Take 5-bromo-2,3-dihydrobenzofuran (1eq), NiCl2·(H2O) 1.7 (0.15eq, obtained by heating and dehydrating NiCl2·(H2O)6), 1,3-dicyclohexyl imidazole tetrafluoroborate (0.15eq), tert-butyl magnesium chloride (2eq, 1.7M in THF) was slowly added dropwise with stirring at -10°C, and the reaction was carried out at low temperature for 3h. After the reaction was completed by TLC detection, saturated ammonium chloride was quenched, and the mixture was extracted with ethyl acetate three times (10mL*3). The organic phases were combined, washed with saturated brine (10mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 5-tert-butyl-2,3-dihydrobenzofuran.

[0106] 5-tert-Butyl-2,3-dihydrobenzofuran (1 eq) was dissolved in dichloromethane and stirred at 0°C for 10 min. Chlorosulfonic acid (3 eq, 1.5 M in DCM) was added dropwise and the mixture was warmed to room temperature and monitored by TLC. After completion of the reaction, the mixture was poured into ice-cold brine to quench the reaction. The mixture was then extracted three times with dichloromethane (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the sulfonyl chloride intermediate. The sulfonyl chloride compound (1 eq) was then dissolved in acetone and ammonia (5 eq) was added with stirring at room temperature. The mixture was reacted at room temperature for 2 h. After completion of the reaction by TLC, the reaction solution was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the sulfonamide intermediate.

[0107] The target sulfonamide fragment (1 eq) was dissolved in THF, and NaH (1.2 eq) was added at 0°C. Intermediate 1 (1.2 eq) was then added with stirring. The mixture was reacted at room temperature for 2 h. After completion of the reaction by TLC, the mixture was extracted with ethyl acetate three times (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the final product.

[0108] Example 5: Preparation of sulfonylurea compound N77

[0109] 5-Amino-2,3-dihydrobenzofuran (1 eq) was dissolved in dichloromethane and stirred at 0°C for 10 min. Chlorosulfonic acid (3 eq, 1.5 M in DCM) was added dropwise. The reaction was warmed to room temperature and monitored by TLC. After completion, the reaction was poured into ice-cold brine to quench. The mixture was then extracted three times with dichloromethane (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the sulfonyl chloride intermediate. The sulfonyl chloride compound (1 eq) was then dissolved in acetone, and ammonia (5 eq) was added with stirring at room temperature. The reaction was allowed to react at room temperature for 2 h. After completion of the reaction by TLC, the reaction solution was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the sulfonamide intermediate.

[0110] Then, hydrogen peroxide (14 eq) was added dropwise to a mixture of the above-mentioned sulfonamide raw material (1 eq) and diphenyl diselenide (0.2 eq). The mixture was reacted at room temperature for 2 h. After TLC detection, the reaction was quenched with water and extracted with ethyl acetate three times (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the oxidized sulfonamide intermediate.

[0111] The target sulfonamide fragment (1 eq) was dissolved in THF, and NaH (1.2 eq) was added at 0°C. After stirring for 10 min, intermediate 1 (1.2 eq) was added and reacted at room temperature for 2 h. After TLC detection, the reaction was completed and extracted with ethyl acetate three times (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the final product.

[0112] Example 6: Preparation of nitrosulfonylurea compounds N78-N87

[0113] A sulfonyl chloride compound (1 eq) was dissolved in 1,4-dioxane. A solution of ammonia in dioxane (5 eq) was added with stirring at room temperature. The mixture was allowed to react at room temperature for 2 h. After TLC, the reaction mixture was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the sulfonamide intermediate. The sulfonamide intermediate (1 eq) was then dissolved in EtOH, 3-chloropropylamine hydrochloride (1.1 eq) and DIPEA (2.2 eq) were added, and the mixture was refluxed at 90°C for 1 h. After TLC, the reaction mixture was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the mesochloropropylamine intermediate.

[0114] The chloropropylamine intermediate compound (1 eq) was dissolved in DMF, potassium carbonate (3 eq) and KI (0.01 eq) were added, and the mixture was stirred at 100°C for activation. Secondary amine (2 eq) was then slowly added dropwise and stirred for 2 h. After the reaction was completed by TLC, the mixture was washed with a large amount of water (50 mL*3) and extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the target sulfonamide fragment.

[0115] The target sulfonamide fragment (1 eq) was dissolved in THF, and NaH (1.2 eq) was added at 0°C. After stirring for 10 min, intermediate 1 (1.2 eq) was added and allowed to react at room temperature for 2 h. After TLC, the reaction was completed and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the final product. Alternatively, if a solid precipitated after the reaction, the resulting solid was filtered and slurried / recrystallized with an appropriate solvent to obtain the final product.

[0116] Example 7: Preparation of sulfonylurea compounds S08-S16

[0117] A sulfonyl chloride compound (1 eq) was dissolved in 1,4-dioxane, and a solution of ammonia in dioxane (5 eq) was added under stirring at room temperature. The mixture was reacted at room temperature for 2 h. After the reaction was completed by TLC, the reaction solution was concentrated under reduced pressure and extracted three times with ethyl acetate (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain the sulfonamide intermediate.

[0118] The sulfonamide intermediate compound (1 eq) was dissolved in DMF, potassium carbonate (3 eq) and secondary amine (2 eq) were added, and the mixture was stirred at 90°C for 4 h. After the reaction was completed by TLC, the mixture was washed with a large amount of water (50 mL*3) and extracted with ethyl acetate three times (10 mL*3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the target sulfonamide fragment.

[0119] The target sulfonamide fragment (1 eq) was dissolved in THF, and NaH (1.2 eq) was added at 0°C. After stirring for 10 min, intermediate 1 (1.2 eq) was added and allowed to react at room temperature for 2 h. After completion of the reaction, the mixture was extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain the final product. If a solid precipitated after completion of the reaction, the resulting solid was filtered and slurried / recrystallized with an appropriate solvent to obtain the final product.

[0120] Example 8: Determination of biological activity

[0121] 1. Experimental reagents, materials and main instruments

[0122] 1. Reagents and Materials: Dimethyl sulfoxide (Aladdin, 67-68-5), RPMI 1640 (VM-2101BM, Vistech), Dulbecco's Modified Eagle's Medium (Vistech, VM-1101BM), Opti-MEM medium (Gibco, 31985062), fetal bovine serum (FBS, Vistech, SE100-011), penicillin-streptomycin solution (PS, Gibco, 15140-122), macrophage colony-stimulating factor (M-CSFB, Bio-techne, 416-ML), lipopolysaccharide (LPS, MedChemExpress, HY-D1056), nigericin (Nigericin, MedChemExpress, HY-127019), Mouse IL-1 beta / IL-1F2 Quantikine HS ELISA Kit (R&D Systems, MHSLB00), Mouse IL-1 beta / IL-1F2 DuoSet ELISA (R&D Systems, DY401-05), ethanol (Shanghai Trial, 10009159), RIPA lysis buffer (Beyotime, P0013B), chloroform (Shanghai Trial, 10006818), methanol (Shanghai Trial, 10014128), NLRP3 antibody (Cell Signaling Technologies, 15101), Caspase-1 antibody (Cell Signaling Technologies, 24232), IL-1β antibody (Cell Signaling Technologies, 12426), GAPDH antibody (Cell Signaling Technologies, 2118), HRP-Goat Anti-rabbit IgG (H+L) (Bioker, BK-R050), and ECL exposure solution (NCM Biotech, P2300).

[0123] 2. Experimental instruments: microplate reader (Tecan Group Ltd., Swiss), water bath (Shanghai Boxun Medical Biological Instrument Co., Ltd., DK-8D), high-speed centrifuge (Eppendorf AG, Centrifuge 5420), protein electrophoresis instrument (Bio-Rad Laboratories Co., Ltd., Tetra Blotting Module), exposure instrument (Ebiotrade, e-Blot).

[0124] 3. Experimental consumables: dissecting scissors, cell scraper, 12-well cell culture plate (Corning, 3737), 24-well cell culture plate (Corning, 3738), disposable bacterial culture dish (Bbi-life sciences, F611004).

[0125] 4. Experimental Animals: Male ICR mice (6-12 weeks) were purchased from the Laboratory Animal Center of Hangzhou Normal University and maintained in a constant temperature and humidity environment. All animal husbandry and experimental use were in accordance with the "Guide for the Care and Use of Laboratory Animals."

[0126] 2. Experimental methods:

[0127] 1. Isolation and Differentiation of BMDM Cells

[0128] 1) Removing Mouse Leg Bones: Mice were sacrificed by dislocation. The legs and back were disinfected with ample 75% ethanol. The hind limbs were then dissected along the greater trochanter at the base of the thigh using scissors. The muscle tissue was removed and the bones were placed in cold PBS. (This step was performed in a cleanroom; subsequent steps were transferred to the cell room.)

[0129] 2) BMDM Cell Isolation and Induction: Rinse the leg bones with PBS to remove excess muscle, rinsing approximately 2-3 times. Use scissors to cut the femur and tibia at both ends. Use a 1 mL syringe to draw up cold induction medium and aspirate the bone marrow from the femur and tibia. Repeat this aspiration three times until no distinct red color is visible within the leg bones. Use a pipette to repeatedly pipette the medium containing the bone marrow cells to disperse cell clumps. Transfer the medium to a 15 mL centrifuge tube and centrifuge at 1500 rpm for 5 minutes. Discard the supernatant and resuspend in approximately 1-2 mL of red blood cell lysis buffer. Use a pipette to disperse the cells into single cells. After standing at room temperature for 5 minutes, add 10 mL of culture medium to terminate the reaction. Filter the medium through a 0.45 μm cell filter and centrifuge at 1500 rpm for 5 minutes. Discard the supernatant, resuspend the cells, and count the cells. Resuspend the cells in 1640 medium (10% FBS, 2% PS) supplemented with 20 ng / mL M-CSF and transfer the cell suspension to a non-treated disposable bacterial culture dish for culture.

[0130] 2. Activation of the canonical NLRP3 inflammasome

[0131] Use 4-6 days of well-differentiated BMDM cells, discard the culture supernatant, and add an appropriate amount of PBS to wash the cells. Add 10 mL of DMEM medium and scrape the cells with a cell scraper. Resuspend the cells and transfer them to a centrifuge tube. Centrifuge at 1500 rpm for 10 minutes. Remove the supernatant, resuspend the BMDM cells in DMEM medium, and divide them into 12-well plates, so that the number of cells per well is approximately 5 × 10 5 / mL. The next day, 500 μL of Opti-MEM medium (containing 500 ng / mL LPS) was added to each well of BMDM cells for pretreatment for 4-5 hours. The compound was weighed using an analytical balance and placed in a sterile centrifuge tube. Dimethyl sulfoxide (DMSO) was then added to dissolve the compound to the desired concentration. Drug treatment was added for 30 minutes, followed by stimulation with 10 μM Nigericin for 60 minutes. The cell supernatant (SN) and cell lysate (Input) were collected. Western Blot analysis and ELISA analysis were performed based on the obtained cell supernatant (SN) and cell lysate (Input).

[0132] 3. Protein Extraction and Sample Preparation

[0133] 1) Protein extraction from cell culture supernatant: Place the sample in a centrifuge and centrifuge at 13,000 rpm for 5 minutes to remove dead cells and impurities. Transfer the supernatant to a new EP tube, add 500 μL of methanol and 125 μL of chloroform, vortex evenly, and centrifuge at 13,000 rpm for 5 minutes. Remove the upper liquid layer, which now represents the protein solution. Add 500 μL of methanol, vortex evenly, and centrifuge at 13,000 rpm for 5 minutes. The protein precipitates at the bottom of the EP tube. Remove the supernatant and let it stand for 10 minutes to evaporate the methanol. Add 60 μL of 1× SDS loading buffer and fully dissolve the protein. Denature at 100°C for 10 minutes.

[0134] 2) Extraction of cell proteins: After removing the supernatant, add 120 μL of cell lysis buffer to the cells. After fully lysing the cells, add 5× SDS loading buffer and denature at 100°C for 10 min.

[0135] 4. Western Blot Detection

[0136] Western blotting was performed on a 4-12% protein gel. Electrophoresis was performed at a voltage of 70-80V for 20-30 minutes, and the separation gel was run at 130-160V for 60-90 minutes. Activate the PVDF membrane in methanol for 5-10 minutes. Pre-soak the filter paper and sponge in transfer buffer. After electrophoresis, soak the protein gel in transfer buffer for 5-10 minutes. Transfer the membrane using a sandwich structure of sponge-filter paper-gel-membrane-filter paper-sponge at 250mA for 90 minutes. Block with 5% skim milk for 1 hour. Incubate with the primary antibody diluted in 5% BSA, depending on the antibody dilution, overnight at 4°C. The next day, wash three times with 1× TBST, 5 minutes apart. Incubate with the secondary antibody diluted in 1× TBST, 1 hour apart, and wash three times with 1× TBST, 5 minutes apart. After washing, expose the membrane with ECL ultrasensitive exposure solution.

[0137] The inhibitory effect of the compound on the NLRP3 inflammasome pathway was evaluated by measuring the amount of IL-1β secreted by BMDM stimulated by LPS+Nigericin after compound treatment. The grayscale value of the IL-1β band in the culture supernatant and the internal reference protein GAPDH in the cell lysate were calculated by ImageJ and their ratio was calculated. The grayscale value of the control group (DMSO treatment group) was used for normalization, and the IC was calculated using GraphPad Prism. 50 .

[0138] In this example, MCC-950, a compound with better activity in the prior art, was used as a reference to test the inhibitory activity of the new compound on the NLRP3 inflammasome.

[0139] The above experimental results are detailed in the table below.

[0140] 3. Experimental Results

[0141] The biological activities of the compounds of the present invention were determined by the above experiments. All compounds had different degrees of inhibitory effects on NLRP3. The results are shown in the table below. Among them, the IC values ​​of the compounds designated as "A" for NLRP3 inhibitory activity were 50 The value is IC 50 <0.1 μM; compounds designated as "B" provided IC 50 Value is 0.1μM≤IC 50 <1.0 μM; compounds designated as "C" for activity provided IC 50 Value is 1.0μM≤IC 50 <10μM.

[0142] Among them, the compound designated as "A" for NLRP3 inhibitory activity has a specific IC 50 See the table below for values.

[0143] Example 9 Effects of the sulfonylurea compounds of the present invention on gout mice

[0144] 1. Experimental Animals, Reagents, and Main Instruments

[0145] 1. Experimental Animals: Male ICR mice (5-8 weeks, ~25 g / mouse) were purchased from the Laboratory Animal Center of Hangzhou Normal University and housed in a constant temperature and humidity environment. All animal husbandry and experimental use were in accordance with the "Guide for the Care and Use of Laboratory Animals."

[0146] 2. Experimental reagents: uric acid (purchased from Sigma-Aldrich), compound solvent (5% DMSO + 45% PEG400 + 20% DMA + 20% glycerol + 10% ethanol).

[0147] 3. Experimental instruments: Von Frey needle prick pain test kit (Von Frey needle prick pain test kit, Aesthesio; animal test cage; provided by the Experimental Animal Center of Hangzhou Normal University); digital display vernier caliper.

[0148] 2. Experimental methods:

[0149] 1. Preparation of Sodium Urate Crystals: Dissolve 1.68 g of uric acid in 500 mL of deionized water, heat to 70°C, and adjust the pH to 7.2-7.4 with 0.1 M NaOH. Recover MSU crystals by centrifugation, wash with ethanol, and dry at 55°C for 2 h. MSU crystals are sterilized and then resuspended in PBS to 25 mg / mL before each experiment.

[0150] 2. Mouse Gout Model: Mice were acclimated for one week and divided into 10 groups of 6-7 mice each. They were orally administered with the drug (3 mg / kg). 0.5 h after administration, MSU (50 μL, 25 mg / mL) was injected into the right footpad to induce gout, and PBS was injected into the left footpad. 8 h later, the drug was administered orally again. 0.5 h after administration, MSU and PBS were injected into the corresponding footpad again to induce gout. Before modeling, the initial mechanical pain sensation (Von Fery) values ​​of the left and right hind footpads of the mice were measured, and the initial footpad thickness was measured. 24 h after the first administration, the mechanical pain sensation (Von Fery) values ​​of the left and right hind footpads of the mice were measured, the footpad thickness was measured, and footpad tissue was extracted.

[0151] 3. Experimental Results

[0152] Compound N21 Na + (Original number M48Na + ) on relevant validation parameters of the MSU-induced gout model in mice are shown in Figure 1: (A) Foot swelling: higher values ​​indicate greater footpad swelling; (B) 50% mechanical withdrawal threshold: higher values ​​indicate less pain; (C) IL-1β level: higher values ​​indicate a greater inflammatory response. Number of animals per group: n = 6-7. Statistical analysis: *p < 0.05, **p < 0.01, ****p < 0.0001.

[0153] The experimental results showed that when the drug concentration was 3 mg / kg, compound N21 Na + Compared with the first-line drugs colchicine and indomethacin, compound N21 Na has similar efficacy.+ The efficacy of the compound N21 Na is better than that of the first-line drug indomethacin and similar to that of the first-line drug colchicine. Compared with the control group (solvent group), there is a statistically significant difference. + It can significantly alleviate the occurrence of MSU-induced gout in the feet of mice.

[0154] Example 10 Safety Evaluation of Sulfonylurea Compounds of the Present Invention

[0155] 1. Experimental Animals, Reagents, and Main Instruments

[0156] 1. Experimental Animals: Male and female ICR mice (3-5 weeks old) were purchased from the Laboratory Animal Center of Hangzhou Normal University and housed in a constant temperature and humidity environment. All animal husbandry and experimental use were in accordance with the "Guide for the Care and Use of Laboratory Animals."

[0157] 2. Experimental reagents: Compound N21 Na + (Original number M48 Na + ) / MCC950 (100 or 200 mg / kg), solvent (5% DMSO, 20% DMA, 45% PEG 400, 20% glycerol, 10% ethanol).

[0158] 2. Experimental Methods

[0159] 1. Mouse treatment: Male and female ICR mice (3-5 weeks old) were gavaged with N21 Na daily. + MCC950 (100 or 200 mg / kg) or solvent (5% DMSO, 20% DMA, 45% PEG 400, 20% glycerol, 10% ethanol) was administered for 14 days. There were 4 mice in each group.

[0160] 2. Body weight monitoring: Measure the body weight of mice before each administration.

[0161] 3. Serum analysis: Serum was collected on the 14th day of the experiment and sent to the Animal Center of Hangzhou Normal University for analysis of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP) and creatinine (CR).

[0162] 4. Organ processing: Remove the heart, liver, spleen, lungs, and kidneys, and record the organ morphology and weight.

[0163] 5. Histological examination: Pathological analysis was performed by Bios Biotechnology.

[0164] 3. Experimental Results

[0165] The experimental results show that N21 Na + (Original number M48 Na +) showed no obvious toxic reactions in mouse body weight, serum indicators and organ morphology, and HE staining showed no significant damage to the liver and kidneys, indicating good safety.

[0166] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound of formula 1, or a tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt thereof, Where, R1 is selected from: H, halogen, cyano, hydroxy, substituted or unsubstituted C 1-3 Amide, substituted or unsubstituted sulfonamide, substituted or unsubstituted C 1-3 Acyl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted C 5-10 aryl; R2 is selected from substituted or unsubstituted C 5-16 Aryl or heteroaryl or cycloalkyl or heterocycloalkyl.

2. The compound according to claim 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, wherein: R2 is selected from the group consisting of substituted or unsubstituted pyrazolyl, substituted or unsubstituted furanyl, substituted or unsubstituted tetrahydrofuranyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted pyranyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted triazolyl, substituted or unsubstituted tetrazolyl, substituted or unsubstituted imidazolyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted morpholinyl, substituted or unsubstituted piperazinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted phenyl, substituted or unsubstituted phenylheteroaryl, substituted or unsubstituted phenylheterocyclyl, substituted or unsubstituted biphenyl, substituted or unsubstituted quinolyl, substituted or unsubstituted isoquinolyl, substituted or unsubstituted naphthyl, substituted or unsubstituted pyrazinyl and substituted or unsubstituted pyrimidinyl.

3. The compound according to claim 2, or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, characterized in that: R2 is selected from the group consisting of: substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted quinolyl, substituted or unsubstituted quinoxalinyl, substituted or unsubstituted phenylheteroaryl, substituted or unsubstituted phenylheterocyclyl, substituted or unsubstituted biphenyl, substituted or unsubstituted thienyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted pyrrolidinyl.

4. The compound according to claim 3, or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, characterized in that: R2 is R3 and R 31 independently selected from: H, hydroxy, halogen, cyano, nitro, substituted or unsubstituted 5-7 membered heterocyclic ring containing 1-2 heteroatoms independently selected from N, O or S, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 5- 10 Aryl or heteroaryl containing 1-2 heteroatoms independently selected from N, O or S, a sulfonamide group containing 1-2 heteroatoms independently selected from N, O or S; Alternatively, two adjacent R3 together with the carbon atoms to which they are attached form a 3-10 membered saturated ring, a substituted or unsubstituted C 5-10 Aromatic ring (e.g. benzene ring) or 3-10 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S or C 5-10 aromatic heterocycles; x = 1, 2, 3, 4, or 5; y = 1, 2, 3, or 4; R4 is selected from the group consisting of: H, OH, substituted or unsubstituted amino, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted C 5-10 Aryl, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S; R5 is selected from: H, substituted or unsubstituted C 1-3 Acyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 5-10 Aryl, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S; R6 and R7 are independently selected from: H, substituted or unsubstituted C 1-3 Acyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 5-10 Aryl, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted 3-10 membered heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S; n and m are independently selected from 0, 1, 2, 3, 4, 5 or 6.

5. The compound according to claim 4, or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, characterized in that: R1 is selected from: H, halogen, cyano; R2 is R3 is selected from: halogen, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-4 Alkoxy, substituted or unsubstituted C 5-10 Aryl (e.g. phenyl), -substituted or unsubstituted C 1-4 Alkyl-OH, -N(R 61 )-substituted or unsubstituted C 1-6 Alkyl-NR 62 R 72 , -O-substituted or unsubstituted C 1-6 Alkyl-NR 62 R 72 , Alternatively, two adjacent R3 together with the carbon atom to which they are attached form a substituted or unsubstituted C 5-10 an aromatic ring (e.g., a benzene ring) or a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S; x = 1, 2, or 3; R 61 Selected from: H, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-3 acyl group; R 62 and R 72 Independently selected from: H, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-3 Acyl, or R 62 and R 72 Together with the nitrogen atom to which they are attached, they form a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S.

6. The compound according to claim 4, or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, wherein: R1 is selected from: H; R2 is R3 is selected from: halogen (such as Br), nitro, substituted or unsubstituted C 1-6 Alkyl, -N(R 61 )-substituted or unsubstituted C 1-6 Alkyl-NR 62 R 72 , Alternatively, two adjacent R3 together with the carbon atom to which they are attached form a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from N, O or S; x = 1, 2, or 3; R 61 Selected from: H, substituted or unsubstituted C 1-4 alkyl; R 62 and R 72 Independently selected from substituted or unsubstituted C 1-6 Alkyl; or, R 62 and R 72 Together with the nitrogen atom to which they are attached, they form a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1, 2 or 3 heteroatoms independently selected from N, O or S.

7. A compound selected from the group consisting of: a tautomer, a mesomer, a racemate, an enantiomer, a diastereomer, a mixture, or a pharmaceutically acceptable salt thereof:

8. The compound according to claim 7, or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, wherein: The compound is selected from the group consisting of: Preferably, the compound is selected from the group consisting of:

9. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the compound according to any one of claims 1 to 8 or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt, and optionally a pharmaceutically acceptable carrier.

10. Use of the compound according to any one of claims 1 to 8 or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture or pharmaceutically acceptable salt in the preparation of an NLRP3 inhibitor.

11. The use according to claim 10, characterized in that The NLRP3 inhibitor is a drug for treating and / or preventing NLRP3-related diseases.

12. The use according to claim 11, characterized in that The NLRP3-related diseases include but are not limited to gout, neurodegenerative diseases, non-alcoholic steatohepatitis, type II diabetes, inflammatory bowel disease, hepatitis, and arthritis.

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