Benzothiophene deuterated compound and use thereof

Abstract

The present invention belongs to the field of chemical and pharmaceutical technologies. Disclosed in the present invention are a benzothiophene deuterated compound and the use thereof. The benzothiophene deuterated compound as represented by formula I provided by the present invention can be used as an NLRP3 inhibitor, has high activity and excellent pharmacokinetic properties, and provides a new path for treating NLRP3-related diseases.

Description

Benzothiophene deuterated compounds and their uses Technical Field

[0001] This invention belongs to the field of chemical medicine and relates to a class of benzothiophene deuterated compounds and their uses. Background Technology

[0002] Inflammasomes are protein complexes that recognize intracellular pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs). Inflammasome assembly triggers proteolysis, cleaving dormant procaspase-1 into active caspase-1, and converting the cytokine precursors pro-IL-1β and pro-IL-18 into mature, biologically active IL-1β and IL-18, respectively. This process regulates the expression of inflammation-related genes, leading to various biological effects. As receptors for the body's innate immunity, inflammasome activation can resist pathogen infection and stress damage; however, uncontrolled activation can amplify inflammatory effects and cause organ damage. Currently, research on inflammasomes containing pyrin domain-containing protein 3 (NLRP3), a member of the nucleotide-binding oligomerization domain (NOD)-like receptor family, is the most popular.

[0003] The NLRP3 inflammasome consists of a sensor (NLRP3), an adapter (ASC, also known as PYCARD), and an effector (caspase 1). Classical NLRP3 inflammasome activation is triggered by the co-stimulation of two signals. The first signal activates the TLR4 (Toll-like receptor 4) signaling pathway, promoting NF-κB nuclear translocation, inducing the production of precursors such as IL-1β and IL-18, and inducing post-translational modifications of NLRP3. The second signal promotes the formation of the NLRP3 / ASC / pro-caspase-1 complex. Upon activation, ASC polymerizes with apoptosis-associated specklike protein containing a cardinolytic activation and recruitment domain. ASC then interacts with cysteine ​​protease caspase-1 to form a complex called the inflammasome. The precursor form of pro-caspase-1 self-cleaves into its activated form. The activated caspase-1 cleaves the precursor forms of pro-inflammatory cytokines IL-1β and IL-18, converting them into their active forms and releasing them extracellularly. This recruits inflammatory cells, amplifying the inflammatory response. The ASC specklike protein can also recruit and activate caspase-8, cleaving the precursor forms of IL-1β and IL-18 into their mature forms and inducing pyroptosis. Non-classical NLRP3 inflammasome activation does not depend on TLR4 signaling pathway activation. Instead, it is initiated by caspase-11 directly recognizing intracellular LPS, which promotes the activation and release of Gasdermin D, thereby mediating cell death.

[0004] Abnormal activation of NLRP3 is associated with many diseases, including inflammasome-related diseases, immunological diseases, inflammatory diseases, neurological diseases, autoimmune diseases and / or autoinflammatory diseases, cancer, chronic metabolic diseases, and neurological diseases. Examples include cryptothermal protein-associated cycle syndrome (CAPS), Mukel-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), neonatal multisystem inflammatory disease (NOMID), familial Mediterranean fever (FMF), nonalcoholic steatohepatitis, alcoholic liver disease, graft-versus-host disease, multiple sclerosis (MS), rheumatoid arthritis, type I / II diabetes and related complications (e.g., nephropathy, retinopathy), psoriasis, Alzheimer's disease, atherosclerosis, gout, chronic kidney disease, sepsis, liver fibrosis, idiopathic pulmonary fibrosis, epilepsy, neuropathic pain, depression, Parkinson's disease, asthma, acute myocardial infarction, lupus erythematosus, rheumatoid arthritis, Crohn's disease, ulcerative colitis, inflammatory bowel disease, rheumatoid arthritis, ankylosing spondylitis, bronchial asthma, acute respiratory distress syndrome, chronic obstructive pulmonary disease, or ischemic stroke. NLRP3 is located upstream of cytokines and can block inflammation at its source, therefore developing new NLRP3 inflammasome inhibitors has high research value. Summary of the Invention

[0005] The purpose of this invention is to develop a class of benzothiophene deuterated compounds and their uses, or their stereoisomers, solvates, metabolites, deuterated products, prodrugs, pharmaceutically acceptable salts or cocrystals, comprising pharmaceutical compositions thereof, for the treatment of NLRP3-related diseases.

[0006] In a first aspect, the present invention provides a compound of Formula I or a pharmaceutically acceptable form thereof, said Formula I having the following structure:

[0007] in,

[0008] Ring A is selected from or (The left-hand bonds of these two structural units are connected to benzothiophene, and the right-hand bonds are connected to L);

[0009] Structural unit Selected from: or

[0010] R1 is selected from C 1～4 Alkyl, hydroxyl substituted C 1～4 Alkyl, C 1～4 Deuterated alkyl, C 1～4 Fluorinated alkyl groups, 3-6 membered cycloalkyl groups, or 3-6 membered fluorocycloalkyl groups;

[0011] R2 is selected from halogen or C. 1～4 alkyl;

[0012] R3 is selected from hydroxyl, amino, or C. 1～4 alkyl;

[0013] R4 is selected from C 1～4 Alkyl or C 1～4 Deuterated alkyl groups;

[0014] The pharmaceutically acceptable form is selected from pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled substances, metabolites, or prodrugs.

[0015] In some embodiments of the present invention, R1 is selected from methyl, hydroxymethyl, deuterated methyl, fluoromethyl, ethyl, hydroxyethyl, isopropyl, cyclopropyl, fluorocyclopropyl, cyclobutyl or fluorocyclobutyl; R2 is selected from fluorine or methyl; R3 is selected from hydroxy, amino or methyl; R4 is selected from methyl or deuterated methyl.

[0016] In some embodiments of the present invention, structural units Selected from: or

[0017] In some preferred embodiments of the present invention, structural units Selected from: or

[0018] This invention also provides specific compounds of Formula I, wherein the compounds are selected from:

[0019] or

[0020] In a second aspect, the present invention provides a pharmaceutical composition having the aforementioned compound of Formula I or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug as the active ingredient, supplemented by a pharmaceutically acceptable carrier.

[0021] A further object of the present invention is to provide a method for preparing the pharmaceutical composition of the present invention, the method comprising combining a compound of formula I or a pharmaceutically acceptable form thereof, or a mixture thereof, with one or more pharmaceutically acceptable carriers.

[0022] The pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention are pharmaceutically acceptable carriers, and examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (2005).

[0023] Pharmaceutical compositions can be administered in any form, as long as they achieve the purpose of preventing, alleviating, preventing, or curing symptoms in human or animal patients. For example, they can be formulated into various suitable dosage forms depending on the route of administration.

[0024] In other embodiments, the administration of the compounds or pharmaceutical compositions of the present invention may be combined with other treatment methods. These other treatment methods may be selected from, but are not limited to, radiotherapy, chemotherapy, immunotherapy, or combinations thereof.

[0025] The present invention also relates to a pharmaceutical formulation having a compound of Formula I above, or a pharmaceutically acceptable form thereof, or a mixture thereof, or a pharmaceutical composition of the present invention as the active ingredient. In some embodiments, the formulation is in the form of a solid dosage form, a semi-solid dosage form, a liquid dosage form, or a gaseous dosage form.

[0026] A further object of the present invention is to provide an article of manufacture, for example, in the form of a kit. The article of manufacture as used herein is intended to include, but is not limited to, medicine boxes and packaging. The article of manufacture of the present invention comprises: (a) a first container; (b) a pharmaceutical composition contained in the first container, wherein the composition comprises: a first therapeutic agent, the first therapeutic agent comprising: a compound of formula I or a pharmaceutically acceptable form thereof, or a mixture thereof; (c) optionally present packaging instructions stating that the pharmaceutical composition may be used to treat a cancerous condition (as defined below); and (d) a second container.

[0027] The first container is a container for containing a pharmaceutical composition. This container may be used for the preparation, storage, transportation, and / or individual / bulk sales. The first container is intended to encompass bottles, jars, vials, flasks, syringes, tubes (e.g. for cream products), or any other container for the preparation, containment, storage, or dispensing of pharmaceutical products.

[0028] The second container is a container for holding the first container and optional instruction manuals. Examples of the second container include, but are not limited to, boxes (e.g., cardboard or plastic boxes), cartons, cartons, bags (e.g., paper or plastic bags), sachets, and cloth bags. The instruction manuals may be physically attached to the outside of the first container by cable ties, glue, U-staples, or other adhesive methods, or they may be placed inside the second container without any physical means of attachment to the first container. Alternatively, the instruction manuals may be located outside the second container. When located outside the second container, it is preferable that the instruction manuals be physically attached by cable ties, glue, U-staples, or other adhesive methods. Alternatively, they may be adjacent to or in contact with the outside of the second container without physical attachment.

[0029] The packaging instructions, such as trademarks, labels, or markings, list information relating to the pharmaceutical composition contained within the first container. The listed information is typically determined by the regulatory authority governing the region where the product is to be sold (e.g., the U.S. Food and Drug Administration). Preferably, the packaging instructions specifically list the approved indications for which the pharmaceutical composition is used. The packaging instructions can be made of any material from which information contained therein or on the material can be read. Preferably, the packaging instructions are made of a printable material (e.g., paper, plastic, cardboard, foil, adhesive paper, or plastic) on which the desired information can be formed (e.g., printed or coated).

[0030] Thirdly, the present invention provides the use of the aforementioned Formula I compound, and related specific compounds or pharmaceutically acceptable forms thereof, or the use of the pharmaceutical compositions of the present invention in the preparation of NLRP3 inhibitors.

[0031] The present invention also provides the use of the aforementioned Formula I compound, and related specific compounds or pharmaceutically acceptable forms thereof, or the use of the pharmaceutical compositions of the present invention in the preparation of medicaments for the prevention or treatment of NLRP3-related diseases.

[0032] The present invention provides a method for preventing or treating NLRP3-related diseases, the method comprising administering to an individual in need a compound of formula I or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention.

[0033] The present invention provides a method for preventing or treating NLRP3-related diseases by combining the above-described Formula I compound or a pharmaceutically acceptable form thereof, or the pharmaceutical composition of the present invention, with other treatment methods, including but not limited to: radiotherapy, chemotherapy, immunotherapy, or combinations thereof.

[0034] In some implementations, the NLRP3-related diseases include: inflammatory diseases, autoimmune diseases, cardiovascular diseases, cancer, kidney diseases, gastrointestinal diseases, respiratory diseases, endocrine diseases, or central nervous system diseases.

[0035] In some implementations, the NLRP3-related diseases include: cryptothermal protein-associated cycle syndrome (CAPS), Mukel-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), neonatal multisystem inflammatory disease (NOMID), familial Mediterranean fever (FMF), nonalcoholic steatohepatitis, alcoholic liver disease, graft-versus-host disease, multiple sclerosis (MS), rheumatoid arthritis, type I / II diabetes and related complications (e.g., nephropathy, retinopathy), psoriasis, Alzheimer's disease, atherosclerosis, gout, chronic kidney disease, sepsis, liver fibrosis, idiopathic pulmonary fibrosis, epilepsy, neuropathic pain, depression, Parkinson's disease, asthma, acute myocardial infarction, lupus erythematosus, rheumatoid arthritis, Crohn's disease, ulcerative colitis, inflammatory bowel disease, rheumatoid arthritis, ankylosing spondylitis, bronchial asthma, acute respiratory distress syndrome, chronic obstructive pulmonary disease, or ischemic stroke.

[0036] In a further preferred embodiment, the compounds of the present invention can be used in combination with radiotherapy, chemotherapy, or immunotherapy to prevent or treat NLRP3-related diseases.

[0037] The beneficial effects of this invention are:

[0038] This invention provides a class of benzothiophene deuterated compounds and their uses. These compounds and compositions can be used to prepare NLRP3 inflammasome inhibitors, providing a new approach for the treatment of NLRP3-related diseases.

[0039] Terminology definition:

[0040] This invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of this invention, except that one or more atoms are replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this invention include (but are not limited to) isotopes of hydrogen (e.g., deuterium). 2 H), tritium ( 3 H); carbon isotopes (e.g., ... 13 C and 14 C); isotopes of chlorine (e.g.) 37 Cl); isotopes of iodine (e.g., Cl); 125 I); nitrogen isotopes (e.g.) 13 N and 15N); isotopes of oxygen (e.g., N); 17 O and 18 O); isotopes of phosphorus (e.g., O); phosphorus isotopes ... 32 P); and isotopes of sulfur (e.g. 34 S).

[0041] In this invention, "polymorph" refers to different solid crystalline phases resulting from the presence of two or more different molecular arrangements in the solid state of certain compounds of this invention. Some compounds of this invention may exist in more than one crystal form, and this invention aims to include various crystal forms and mixtures thereof. Typically, crystallization produces solvates of the compounds of this invention. The term "solvate" as used in this invention refers to an aggregate comprising one or more molecules of the compound of this invention and one or more solvent molecules. The solvent may be water, in which case the solvate is a hydrate. Alternatively, the solvent may be an organic solvent. Therefore, the compounds of this invention can exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvated forms. The compounds of this invention can form true solvates, but in some cases, they may also remain only as indeterminate water or a mixture of water and a portion of indeterminate solvent. The compounds of this invention can react in a solvent or precipitate or crystallize from a solvent. The solvates of the compounds of this invention are also included within the scope of this invention. This invention also covers all possible crystalline forms or polymorphs of the compounds of this invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion.

[0042] In this invention, "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of this invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, and imine-enamine tautomers. It is to be understood that the scope of this invention covers all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0043] In this invention, pharmaceutically acceptable salts include their acid addition salts and base addition salts. Suitable acid addition salts are formed by acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed by bases that form pharmaceutically acceptable salts. A review of suitable salts can be found, for example, in “Remington’s Pharmaceutical Sciences,” Mack Publishing Company, Easton, Pa., (2005); and “Handbook of Pharmaceutical Salts: Properties, Selection, and Use,” Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of this invention are known to those skilled in the art. “Pharmaceutically acceptable acid addition salt” refers to a salt formed with an inorganic or organic acid that retains the bioavailability of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetate, trifluoroacetate, propionate, hexanoate, octanoate, decanoate, undecanoate, glycolate, gluconate, lactate, sebate, adipate, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate, cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, alginate, ascorbate, salicylate, 4-aminosalicylic acid, and naphthalenedisulfonate. These salts can be prepared by methods known in this patent. "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, and aluminum salts. Preferred inorganic salts are ammonium salts, sodium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, and polyamine resins.Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in this patent.

[0044] In this invention, unless otherwise stated, "ester" refers to an ester derived from the compounds described herein, including physiologically hydrolyzable esters (compounds of this invention that can be hydrolyzed under physiological conditions to release free acids or alcohols). The compounds of this invention may themselves be esters.

[0045] The compounds of the present invention can exist as solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be stoichiometric or non-stoichiometric.

[0046] Those skilled in the art will understand that not all nitrogen-containing heterocycles can form nitrogen oxides because nitrogen requires available lone pairs of electrons to be oxidized. Those skilled in the art will identify nitrogen-containing heterocycles capable of forming nitrogen oxides. They will also recognize that tertiary amines can form nitrogen oxides. Synthetic methods for preparing nitrogen oxides of heterocycles and tertiary amines are well known to those skilled in the art, including the oxidation of heterocycles and tertiary amines with peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (mCPBA), hydrogen peroxide, alkyl peroxides such as tert-butyl peroxide, sodium perborate, and dioxiranes such as dimethyldioxirane. These methods for preparing nitrogen oxides have been extensively described and reviewed in the literature, see, for example: T.L. Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp. 748-750 (AR. Katritzky and A.J. Boulton, Eds., Academic Press); and G.W. H. Heeseman and E.S. G. Wierstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp. 390-392 (AR. Katritzky and A.J. Boulton, Eds., Academic Press).

[0047] In this invention, "metabolite" refers to a substance formed in the body upon administration of a compound of the present invention. Metabolites of the compound can be identified using techniques known in the art, and their activity can be characterized by experimental methods. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of the present invention, including compounds obtained by methods that expose the compounds of the present invention to mammals for a time sufficient to produce their metabolites.

[0048] In this invention, a "prodrug" refers to certain derivatives of the compounds of the invention that, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of the invention having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that readily convert in vivo into the desired therapeutically active compounds. Further information regarding the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems," Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella). The prodrugs of the invention can be prepared, for example, by replacing suitable functional groups present in the compounds of the invention with portions known to those skilled in the art as "pro-moiety" (e.g., as described in "Design of Prodrugs," H. Bundgaard (Elsevier, 1985)).

[0049] In this application, "pharmaceutical composition" refers to a formulation of the compounds of the present invention with a medium generally accepted in the art for delivering bioactive compounds to mammals (e.g., humans). This medium includes pharmaceutically acceptable carriers. The purpose of the pharmaceutical composition is to facilitate administration to the organism, enhance the absorption of the active ingredient, and thereby exert its bioactivity.

[0050] In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, flow aid, sweetener, diluent, preservative, dye / coloring agent, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that is permitted by the relevant government regulatory authority or is acceptable for human or livestock use.

[0051] As used herein, the terms “drug combination,” “drug co-administration,” “combination therapy,” “administration of other treatments,” and “administration of other therapeutic agents” refer to pharmaceutical treatments achieved by mixing or combining more than one active ingredient, including fixed and non-fixed combinations of active ingredients. The term “fixed combination” refers to the simultaneous administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity or dosage form. The term “non-fixed combination” refers to the simultaneous, combined, or sequential administration to a patient of at least one compound described herein and at least one synergistic agent in the form of a single entity at variable intervals. These also apply to cocktail therapies, such as the administration of three or more active ingredients.

[0052] In this invention, unless otherwise stated, "tumor" includes, but is not limited to, diseases such as leukemia, gastrointestinal stromal tumor, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, squamous cell carcinoma of the lung, adenocarcinoma of the lung, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, cervical cancer, ovarian cancer, intestinal cancer, nasal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, and oral cancer.

[0053] In this invention, unless otherwise stated, "treatment" means reversing, alleviating, or inhibiting the progression of a disease or condition or one or more symptoms of such a disease or condition, or preventing such a disease or condition or one or more symptoms of such a disease or condition.

[0054] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention. Detailed Implementation

[0055] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in the art or in accordance with the product manual.

[0056] The reagents and raw materials used in the embodiments of this invention are all commercially available.

[0057] Table 1. Abbreviations and their meanings in this invention.

[0058] The structure of the compound described in this invention was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). Chemical shifts were expressed as 10⁻⁶.-6 (ppm) is given as the unit.

[0059] MS measurements were performed using an Agilent SQD (ESI) mass spectrometer (manufacturer: Agilent, signal: 6110).

[0060] HPLC determinations were performed using an Agilent 1200DAD high-performance liquid chromatograph (Sunfirc C18, 150X 4.6mm, 5µm column) and a Waters 2695-2996 high-performance liquid chromatograph (Gimini C18, 150X 4.5mm, 5µm column).

[0061] The silica gel plates used for thin-layer chromatography are Qingdao Ocean GF254 silica gel plates. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15mm-0.2mm, while the silica gel plates used for thin-layer chromatography separation and purification of products have a diameter of 0.4mm-0.5mm.

[0062] Column chromatography typically uses Qingdao Marine 100-200 or 200-300 mesh silica gel as a carrier.

[0063] Unless otherwise specified in the following examples, all reactions are carried out under an argon or nitrogen atmosphere. An argon or nitrogen atmosphere refers to a reaction flask connected to an approximately 1L argon or nitrogen balloon. A hydrogen atmosphere refers to a reaction flask connected to an approximately 1L hydrogen balloon. Hydrogenation reactions are typically performed under vacuum, followed by hydrogen filling, and repeated three times.

[0064] Intermediate INT1: 2-(4-(methoxymethoxy)benzo[b]thiophene-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane

[0065] Step 1: CuBr2 (146.5 g, 656 mmol) was added to EtOAc (250 mL), and stirred at 80 °C for 10 minutes. Then, compound INT1a (25.0 g, 164 mmol) was dissolved in chloroform (250 mL) and added to the suspension. The mixture was refluxed at 80 °C overnight. After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was slurried with EtOAc (500 mL) for 0.5 h, filtered, and the filtrate was concentrated to dryness to obtain the target compound INT1b (46.0 g, 148 mmol, light brown solid, 90% yield), MS: [M+H]. + =309.0,311.0,313.0.

[0066] Step 2: Compound INT1b (45.0 g, 145 mmol) and Li₂CO₃ (26.8 g, 363 mmol) were added to DMF (450 mL) and stirred at 100 °C for 6 h. After the reaction was complete, the mixture was filtered, and the filtrate was treated with hydrochloric acid aqueous solution (900 mL, 0.5 N), extracted with EtOAc (400 mL × 2), washed with water (300 mL × 2), dried over anhydrous Na₂SO₄, filtered, and concentrated to obtain the target compound INT1c (31.0 g, 135 mmol, light brown solid, yield 93%). MS: [MH] - =227.0,229.0.

[0067] Step 3: Compound INT1c (220 g, 960 mmol) and diisopropylethylamine (186 g, 1440 mmol) were added to dichloromethane (1.1 L), and bromomethyl methyl ether (132 g, 1056 mmol) was slowly added dropwise at 0 °C. After the addition was complete, the mixture was stirred at room temperature for 1 h. After the reaction was complete, the mixture was washed with saturated ammonium chloride aqueous solution (300 mL × 3). The organic phase was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure. The solution was separated by column chromatography (PE:EtOAc = 30:1) to obtain the target compound INT1d (183 g, 670 mmol, pale yellow oil, yield 70%). 1H NMR (400 MHz, CDCl3) δ 7.55–7.45 (m, 3H), 7.43 (d, J = 5.5 Hz, 1H), 5.27 (s, 2H), 3.70 (s, 3H).

[0068] Step 4: Compound INT1d (20.0 g, 73.2 mmol), bis-pinacol boronic acid ester (22.3 g, 87.8 mmol), KOAc (14.3 g, 146 mmol), and Pd(PPh3)Cl2 (2.57 g, 3.66 mmol) were added to anhydrous dioxane (200 mL), and reacted at 100 °C for 12 h under nitrogen protection. After the reaction, the mixture was filtered, the filtrate was concentrated under reduced pressure and evaporated to dryness, and separated by column chromatography (PE:EtOAc = 30:1) to obtain the target compound INT1 (14.5 g, 45.3 mmol, colorless solid, yield 62%). 1 ¹H NMR (400MHz, CDCl₃) δ 7.71 (d, J = 8.1 Hz, 1H), 7.63 (d, J = 8.1 Hz, 1H), 7.55 (d, J = 5.5 Hz, 1H), 7.35 (d, J = 5.6 Hz, 1H), 5.25 (s, 2H), 3.62 (s, 3H), 1.37 (s, 12H). Intermediate INT₂: (3R, 5R)-5-fluoro-1-methylpiperidin-3-amine

[0069] Step 1: Compound INT2a (4.75 g, 21.8 mmol), paraformaldehyde (1.31 g, 43.6 mmol), and acetic acid (0.26 g, 4.36 mmol) were added to methanol (50 mL), followed by sodium cyanoborohydride (2.74 g, 43.6 mmol). The mixture was stirred at 50 °C for 1 h. After the reaction was complete, silica gel was added, the mixture was stirred, and the solution was evaporated to dryness. The solution was then purified by column chromatography (DCM:MeOH = 10:1, iodine indicator) to give compound INT2b (4.67 g, 20.1 mmol, yield 92%). 1 H NMR (400MHz, CDCl3-d) δ4.92-4.64(m,2H),3.99(s,1H),2.77-2.39(m,3H),2.30(s,3H),1.94-1.80(m,1H),1.45(s,9H); MS / ESI[M+H] + =233.0.

[0070] Step 2: Compound INT2b (4.67 g, 20.1 mmol) was dissolved in dioxane (25 mL), and then HCl / dioxane (25 mL, 101 mmol, 4.0 M) was added. The mixture was stirred at 25 °C for 1 h. After the reaction was completed, the product INT2 (4.60 g, 22.4 mmol, yield 111%) was obtained by evaporation under reduced pressure. 1 H NMR (400MHz, CD3OD-d4) δ5.37-5.24(m,1H),4.98-4.89(m,1H),3.92-3.77(m,3H),3.51( dd,J=39.2,13.9Hz,1H),3.06(s,3H),2.63-2.55(m,1H),2.15-1.98(m,1H); MS / ESI[M+H] + =133.0.

[0071] Example 1: 6-(((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)amino)-3-(4-hydroxybenzo[b]thiophene-5-yl)-4-(methyl-d3)-1,2,4-triazine-5(4H)-one

[0072] Step 1: Compound 1a (4 g, 20.84 mmol) was placed in a reaction flask, DMF (40 mL) was added, followed by DIEA (3.23 g, 25.00 mmol), and then deuterated iodomethane (3.02 g, 20.84 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC until complete. No purification was performed before proceeding to the next step. MS: [M+H] + :208.1.

[0073] Step 2: Potassium carbonate (2.88 g, 20.84 mmol) and PMBCl (3.92 g, 25.00 mmol) were added sequentially to the reaction solution from Step 1. After the addition was complete, the mixture was heated to 70°C and reacted for 2 hours. The reaction was monitored by TLC until complete. The reaction was quenched with water, extracted with EA, and the organic phase was concentrated. The residue was purified by column chromatography to give a pale yellow solid compound 1c (4 g). MS: [M+H] + :330.2.

[0074] Step 3: Compound 1c (4 g, 12.16 mmol), compound INT2 (1.93 g, 14.60 mmol), Cs2CO3 (11.89 g, 36.48 mmol), BINAP (758 mg, 1.22 mmol), and Pd(OA)2 (274 mg, 1.22 mmol) were added to dioxane (60 mL). The mixture was then purged three times with nitrogen and heated to 110 °C with stirring overnight. After the reaction was complete, the mixture was evaporated to dryness under reduced pressure using silica gel. The solution was then purified by column chromatography (DCM:MeOH = 20:1) to obtain the target compound 1d (3.30 g, pale yellow viscous substance). MS: [M+H] + =381.0.

[0075] Step 4: Compound 1d (3.30 g, 8.68 mmol) was dissolved in dichloromethane (30 mL), and TfOH (3.91 g, 26.05 mmol) was slowly added at room temperature, followed by stirring overnight. After the reaction was complete, the pH was adjusted to approximately 8 with ammonia, dried over anhydrous Na₂SO₄, filtered, and the filtrate was stirred into silica gel and evaporated under reduced pressure. The solution was then separated by column chromatography (DCM:MeOH = 5:1) to obtain the target compound 1e (1.61 g). MS: [M+H] + =261.1.

[0076] Step 5: Add 30 mL of POCl3 to compound 1e (1.61 g, 6.19 mmol), heat to 110 °C and stir overnight. After the reaction is complete, evaporate excess POCl3 to dryness to obtain crude target compound 1f (1.8 g, 2H3PO4 salt, light brown solid, 100% yield). MS: [M+H] + =279.0.

[0077] Step 6: Weigh compound 1f (1.8 g, 3.79 mmol), compound INT1 (1.33 g, 4.17 mmol), Cs₂CO₃ (4.08 g, 12.51 mmol), and Pd(dppf)Cl₂ (279 mg, 0.38 mmol) and add them to dioxane (20 mL) and water (4 mL). Then, purge three times with nitrogen and heat to 100 °C with stirring overnight. After the reaction is complete, evaporate to dryness under reduced pressure using silica gel. Purify by column chromatography (DCM:MeOH = 10:1) to obtain 1 g (814 mg, pale yellow viscous substance) of the target compound. MS: [M+H] + =437.0.

[0078] Step 7: 1 g (814 mg, 1.86 mmol) of compound was placed in a reaction flask, dissolved in methanol (10 mL), followed by 5 mL of hydrochloric acid-methanol solution (4 M). After the addition was complete, the reaction mixture was allowed to react at room temperature for 30 min. After the reaction was complete, the reaction solution was concentrated, and the residue was purified to give compound 1 (pale yellow solid, 443 mg, yield: 59.07%). LC-MS: ESI [M+H] + =393.0; 1 H NMR (400MHz, DMSO) δ13.13(s,1h),7.69(d,J=8.0Hz,1h),7.60(d,J=8.0Hz,1H),7.52(d,J=8.0Hz,1H),7.44(d,J=8.0Hz, 1H),7.30(s,1H),3.99(s,1H),2.35-2.32(m,1H),2.30(s,3H),2.26–2.16(m,1H),1.80-1.73(m,2H),1.55-1.45(m,2H).

[0079] Example 2: (R)-6-((1-ethylpiperidin-3-yl)amino)-3-(4-hydroxybenzo[b]thiophen-5-yl)-4-(methyl-d3)-1,2,4-triazine-5(4H)-one

[0080] Preparation of (R)-6-((1-ethylpiperidin-3-yl)amino)-3-(4-hydroxybenzo[b]thiophene-5-yl)-4-(methyl-d3)-1,2,4-triazine-5(4H)-one: Compound 2 (yellow solid, 37 mg, yield 14.3%) was obtained according to Example 1. LC-MS: ESI [M+H] + =389.0; 1H NMR (400MHz, DMSO) δ7.76(q,J=5.6Hz,2H),7.65(d,J=8.3Hz,1H),7.30(d,J=8.3Hz,1H),3.54(dd,J=25.8,10.7Hz ,2H),3.20(d,J=7.0Hz,2H),2.91–2.75(m,2H),1.99(d,J=10.6Hz,2H),1.84–1.67(m,2H),1.24(t,J=7.2Hz,3H).

[0081] Example 3: (R)-3-((1-ethylpiperidin-3-yl)amino)-6-(4-hydroxybenzo[b]thiophen-5-yl)-4-(methyl-d3)-1,2,4-triazine-5(4H)-one

[0082] Step 1: Compound 1c (4 g, 12.15 mmol), benzyl bromide (2.49 g, 14.58 mmol), palladium acetate (272 mg, 1.22 mmol), BINAP (1.51 g, 2.43 mmol), cesium carbonate (7.92 g, 24.3 mmol), and dioxane (60 mL) were added sequentially to a reaction flask. The mixture was heated to 100 °C and reacted for 8 h. After the reaction was completed, the reaction solution was concentrated, and the residue was purified by column chromatography to obtain compound 3a (yellow oil, 3.8 g, yield 87.96%). LC-MS: ESI [M+H] + =237.0.

[0083] Step 2: Compound 3a (3.8 g, 10.69 mmol) was placed in a reaction flask, followed by the addition of DCM (30 mL), and then trifluoromethanesulfonic acid (4.81 g, 32.08 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC until complete. Ammonia was added to adjust the pH to 8-9. The reaction solution was concentrated, and the residue was purified by column chromatography to obtain compound 3b (pale yellow oil, 1.8 g, yield 71.43%). LC-MS: ESI [M+H] + =236.0.

[0084] Step 3: Compound 3b (1.8 g, 7.66 mmol) was placed in a reaction flask, POCl3 (5 mL) was added, and the mixture was heated to 110 °C and reacted for 18 h. After the reaction was completed, the reaction solution was concentrated to obtain compound 3c (brown oily crude product, 214 mg); LC-MS: ESI [M+H] + =254.0.

[0085] Step 4: Compound 3c (1.9 g, 7.51 mmol) and compound 2a (1.15 g, 9.01 mmol) were placed sequentially in a reaction flask, DMF (10 mL) was added, followed by the slow dropwise addition of DIEA (2.92 g, 22.53 mmol). After the addition was complete, the mixture was heated to 100 °C and reacted for 12 h. After the reaction was completed, the reaction solution was concentrated, and the residue was purified by column chromatography to obtain compound 3d (white solid, 516 mg, yield 19.92%). LC-MS: ESI [M+H] + =346.0.

[0086] Step 5: Place compound 3d (516 mg, 1.49 mmol) in a reaction flask, add Pd / C (50 mg) and MeOH (10 mL), replace with hydrogen, and react at room temperature for 72 h. After the reaction is complete, filter the reaction solution and concentrate to obtain compound 3e (pale yellow oil, 300 mg).

[0087] Step 6: Compound 3e (300 mg, 1.17 mmol) and CuBr (336 mg, 2.35 mmol) were placed in a reaction flask, followed by ACN (10 mL), and then tert-butyl nitrite (242 mg, 2.35 mmol). The mixture was heated to 70°C and reacted for 2 hours. After the reaction was complete, the reaction solution was filtered, concentrated, and the residue was purified by column chromatography to obtain compound 3f (white solid, 140 mg). LC-MS: ESI [M+H] + =320.0.

[0088] Step 7: Compound 3f (140 mg, 0.44 mmol), compound INT1 (169 mg, 0.53 mmol), Pd(dppf)2Cl2 (38 mg, 0.05 mmol), and cesium carbonate (430 mg, 1.32 mmol) were added sequentially to the reaction flask. Then, Diox:H2O (10 mL) was added at a ratio of 4:1. Under nitrogen protection, the mixture was heated to 100 °C and reacted overnight. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by column chromatography and concentrated to give 3 g of compound (brown oil, 42 mg, yield 22.11%). LC-MS: ESI [M+H] + =433.0.

[0089] Step 8: Place 3 g (42 mg, 0.10 mmol) of the compound in a reaction flask, add methanol (10 mL), then slowly add hydrochloric acid-methanol solution (4 M, 6 mL). After the addition is complete, react at room temperature for 1 h. Concentrate the reaction solution, and lyophilize the residue to obtain target compound 3 (yellow solid, 7 mg, yield 19.44%). LC-MS: ESI [M+H] + =389.0; 1H NMR (400MHz, DMSO) δ7.67 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 4.0 Hz, 1H), 7.52 (d, J = 4.0 Hz, 1H), 7.45 (d, J = 8.0 Hz, 1H), 4.08(s,1H),3.10-2.90(m,4H),2.74-2.68(m,2H),1.85-1.80(m,2H),1.65-1.50(m,2H),1.13(t,J=8.0Hz,3H).

[0090] Example 4: 6-(((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)amino)-3-(4-hydroxybenzo[b]thiophen-5-yl)-4-methyl-1,2,4-triazine-5(4H)-one

[0091] Preparation of 6-(((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)amino)-3-(4-hydroxybenzo[b]thiophene-5-yl)-4-methyl-1,2,4-triazine-5(4H)-one was performed according to Example 1. MS / ESI [M+H] + =390.0; 1 H NMR(400MHz, CDCl3-d)δ7.60(d,J=5.4Hz,1H),7.43-7.31(m,2H),7.19(d,J=8.5Hz,1H),6.17(d,J=8.1Hz,1H),4.90-4.72(m,1H),4.45(s,1H) ,3.57(s,3H),2.91-2.78(m,1H),2.68-2.60(m,1H),2.56-2.48(m,1H) ,2.46-2.38(m,1H),2.37(s,3H),2.28-2.17(m,1H),1.91-1.81(m,1H).

[0092] Example 5: (R)-3-((1-ethylpiperidin-3-yl)amino)-6-(4-hydroxybenzo[b]thiophen-5-yl)-4-methyl-1,2,4-triazine-5(4H)-one

[0093] Step 1: Compound 5a (10 g, 57.15 mmol) and cuprous chloride (11 g, 111.11 mmol) were placed in a sealed tube, followed by the addition of tert-butyl nitrite (12 g, 116.50 mmol) and acetonitrile (35 mL). After the addition was complete, the tube was quickly sealed and the mixture was heated to 80 °C and reacted for 4 h. After the reaction was completed, the reaction solution was concentrated, and the residue was purified by column chromatography. The product was eluted with 10% EA:PE, concentrated, and yielded a pale yellow solid product 5b (5.31 g). LC-MS: ESI [M+H] + =193.0.

[0094] Step 2: Compound 5b (5.31 g, 25.24 mmol) was placed in a reaction flask, acetic acid (25 mL) was added, and the flask was placed in an ice bath. Hydrogen peroxide (5 mL) was slowly added dropwise. After the addition was complete, the mixture was slowly brought back to room temperature and reacted overnight. After the reaction was complete, sodium thiosulfate aqueous solution was added to quench the reaction. Extraction was then performed with EA and 5% TEA. The organic phase was concentrated, and the residue was purified by column chromatography to give a pale yellow solid product 5c (5.12 g). LC-MS: ESI [M+H] + =209.1.

[0095] Step 3: Compound 5c (3g, 14.26mmol) was placed in a three-necked flask under nitrogen protection. Anhydrous DMF (30mL) was added, and the reaction was carried out in an ice bath. NaH (1.15g, 28.79mmol) was slowly added. After the addition was complete, the mixture was allowed to return to room temperature for 1 hour. Then, iodomethane (2.02g, 14.26mmol) was slowly added dropwise, and the reaction was allowed to continue overnight at room temperature. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with EA, washed with saturated brine, dried, and concentrated. The residue was purified by column chromatography to give a white solid product 5d (0.8g). LC-MS: ESI [M+H] + =224.0. 1 H NMR (400MHz, DMSO-d6) δ3.45 (s, 3H).

[0096] Step 4: Compound 5d (77 mg, 0.34 mmol), compound INT1 (149 mg, 0.51 mmol), Pd(dppf)Cl2 (13 mg, 0.02 mmol), cesium carbonate (222 mg, 0.69 mmol), dioxane (12 mL), and water (2 mL) were placed in a reaction flask under nitrogen protection and stirred at 100 °C for 15 h. After the reaction was complete, the mixture was cooled to room temperature, and the residue was purified by silica gel column chromatography to obtain compound 5e (90 mg); LC-MS: ESI [M+H] + =338.0.

[0097] Step 5: Compound 5e (90 mg, 0.29 mmol), compound 2a (50 mg, 0.44 mmol), Pd(OAc)2 (6 mg, 0.03 mmol), BINAP (37 mg, 0.06 mmol), Cs2CO3 (190 mg, 0.59 mmol), and 1,4-dioxane (10 mL) were placed in a reaction flask under nitrogen protection and heated to 100 °C for 12 h. After the reaction was complete, the solvent was removed by concentration, and the residue was purified by silica gel column chromatography to obtain the target compound 5f (37 mg); LC-MS: ESI [M+H] + =386.0.

[0098] Step 6: Compound 5f (37 mg, 0.10 mmol) was placed in a reaction flask, DCM (10 mL) was added, and the temperature was lowered to -10 °C. Then, BBr3 (120 mg, 0.48 mmol) was slowly added dropwise. After the addition was complete, the reaction was maintained at -10 °C for 3 h. The reaction was monitored by LCMS until completion. The reaction was quenched with methanol, the reaction solution was concentrated to remove the solvent, and the residue was purified by column chromatography to obtain target compound 5 (22 mg, 31.1%). MS / ESI [M+H] + =386.1. 1 H NMR (400MHz, DMSO) δ8.27(s,1H),7.69(q,J=5.6Hz,2H),7.56(d,J=8.2Hz,1H),7.24(d,J=8.2Hz,1H),3.88-3.79(m,3H),3.04-3.02(m,1H), 2.79-2.76(m,1H),2.41(q,J=7.1Hz,2H),1.96-1.89(m,3H),1.77-1.6 9(m,1H),1.57-1.54(m,1H),1.40-1.37(m,1H),1.02(t,J=7.2Hz,3H).

[0099] Example 6: (R)-6-((1-ethylpiperidin-3-yl)amino)-3-(4-hydroxybenzo[b]thiophen-5-yl)-4-methyl-1,2,4-triazine-5(4H)-one

[0100] The preparation of (R)-6-((1-ethylpiperidin-3-yl)amino)-3-(4-hydroxybenzo[b]thiophen-5-yl)-4-methyl-1,2,4-triazine-5(4H)-one was carried out in accordance with Example 1, yielding compound 6 (70 mg, 80.6%). 1H NMR(400MHz,DMSO-d6)δ10.46(br s,1H),9.60(br s,1H),7.80-7.73(m,2H),7.65(d,J=8.0Hz,1H),7.30(d,J=8.0Hz,1H),4.37-4.27(m,1H),3.58-3.48(m,3H) ,3.29-3.18(m,5H),2.92-2.82(m,1H),2.05-1.96(m,2H),1.88-1.68(m,2H),1.28-1.24(m,3H); MS / ESI[M+H] + =386.0.

[0101] Bioactivity testing:

[0102] 1. Assay of NLRP3 inflammasome inhibitory activity in human monocytes

[0103] Reagents: THP-1 cells: Wuhan Pronosei Biotechnology Co., Ltd., PMA: Sigma-Aldrich, RPMI medium: Hyclone, LPS: Sigma-Aldrich, Opti-MEM medium: Gibco, Nigericin: Invivogen, Human IL-1β ELISA kit: 4A Biotech, Reference compound MCC950: MedChemExpress (MCE).

[0104] Experimental methods: THP-1 cells were cultured in RPMI medium containing PMA (10 μM) at a ratio of 2 x 10⁻⁶ cells / mL. 5 Cells were seeded at a density of 1 μg / mL in 48-well plates and incubated overnight at 37°C with 5% CO2. The next day, the medium was replaced with Opti-MEM medium containing 1 μg / mL LPS. After 3 hours, the drug was added and incubated for 40 minutes; then Nigericin (10 μM) was added and incubated for 40 minutes. The cell supernatant was collected for ELISA analysis. Ref-1 was synthesized according to the method described in Example 26 of patent WO2022238347.

[0105] The experimental results are shown in Table 2 below.

[0106] Table 2. Inhibitory activity of NLRP3 inflammasome

[0107] Conclusion: The compounds of the present invention have good inhibitory activity against NLRP3 inflammasome, and the preferred compounds have better inhibitory effects on NLRP3 inflammasome than reference compound Ref-1.

[0108] 2. Pharmacokinetic Evaluation of the Compound in Balb / c Mice

[0109] Experimental Purpose: To understand the pharmacokinetics of the compound.

[0110] Experimental Basis: Technical Guidelines for Non-clinical Pharmacokinetic Studies of Chemical Drugs, 2014.

[0111] Experimental Scheme: The pharmacokinetics of the compound was investigated by intravenous and intragastric administration to Balb / c mice.

[0112] Sample Preparation: Weigh the compound, dissolve it in DMSO, and then add sodium chloride injection solution to prepare the compound solution for administration.

[0113] Sample Collection: Six Balb / c mice (Chengdu Dashuo Experimental Animal Co., Ltd., License No.: SCXK (Chuan) 2020 - 030), male, 3 were given intravenous injection (IV) and 3 were given intragastric administration (PO). Approximately 0.05 mL of blood was collected at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 24 h, and 48 h after administration. The collected blood was centrifuged at 3500 rpm for 15 min, and the supernatant plasma was collected and stored at -40 °C for further analysis. The plasma drug concentration was quantitatively analyzed by LC-MS / MS, and pharmacokinetic parameters such as peak time (Cmax), area under the plasma concentration-time curve (AUC(0-t)), half-life (T 1 / 2 ), clearance rate (CL), volume of distribution at steady state (Vdss), bioavailability (F), etc. were calculated.

[0114] The results of pharmacokinetic evaluation are shown in Table 3 below:

[0115] Table 3 Pharmacokinetic Test Results of the Compound in Balb / c Mice

[0116] Conclusion: The compound of the present invention has good pharmacokinetic properties in Balb / c mice, including good oral bioavailability, exposure, half-life, and clearance rate, etc. The C max , AUC (0-t) , T 1 / 2 and other pharmacokinetic parameters of compound 3 are significantly better than those of compound 5.

[0117] 3. Evaluation of LPS-induced Mouse Model

[0118] Experimental method: 7-8 week old Balb / c mice were orally administered 15 mg / kg of the compound or a solvent control (sterile 0.9% NaCl solution), followed by intraperitoneal injection of 10 mg / kg LPS (Sigma, L2880) 1 hour later. The survival status of the mice was observed every 12 hours for 72 hours to obtain the 72-hour survival rate.

[0119] The experimental results are shown in Table 4:

[0120] Table 4 shows the survival rate of compounds in LPS-induced mice.

[0121] Conclusion: The compounds of the present invention can increase the survival rate of LPS-induced mice, especially compound 1 at the same dose, which has a better survival rate than compound 4 in LPS-induced mice.

[0122] 5. Evaluation of the distribution of the compound in cerebrospinal fluid and plasma in vivo

[0123] Experimental procedure: Weigh the compound, add a small amount of DMSO, then add sodium chloride solution for injection to prepare a solution of 5 mg / mL. -1 A solution of the compound, ready for administration. For male rats, administer at 5 mg / kg. -1 The drug was administered intravenously. Cerebrospinal fluid and whole blood were collected at 0.25 h and 2 h post-administration, respectively (n=1). Whole blood was centrifuged at 3500 rpm for 15 min, and the supernatant plasma was collected. 10 μL of plasma and 10 μL of cerebrospinal fluid were placed in centrifuge tubes, and 40 μL of a solution containing 20 ng / mL was added. -1 Acetonitrile precipitate of internal standard SAHA, vortex for 30 s, centrifuge at 13000 rpm for 15 min, and collect the supernatant for analysis in a sample vial. Standard curve range: 1–1000 ng / mL -1 .

[0124] Table 5 Results of cerebrospinal fluid and plasma distribution tests in rats after compound administration.

[0125] Conclusion: The compounds of the present invention have good brain penetration potential, especially the cerebrospinal fluid concentration of compound 1 is higher than that of compound 4, and the cerebrospinal fluid concentration of compound 2 is higher than that of compound 6.

Claims

1. A compound of formula I or a pharmaceutically acceptable form thereof, characterized in that: The structure of the formula I is as follows: wherein Ring A is selected from or Structural unit selected from the group consisting of: or R1is selected from C 1～4 alkyl, hydroxy-substituted C 1～4 alkyl, C 1～4 deuteroalkyl, C 1～4 fluoroalkyl, 3- to 6-membered cycloalkyl, or 3- to 6-membered fluorocycloalkyl; R2is selected from halogen or C 1～4 alkyl; R3is selected from hydroxy, amino or C 1～4 alkyl; R4is selected from C 1～4 alkyl or C 1～4 deuteroalkyl; the pharmaceutically acceptable form is selected from a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitroso, isotopically-labeled, metabolite, or prodrug.

2. The compound of claim 1, wherein: R1is selected from methyl, hydroxymethyl, deuterated methyl, fluorinated methyl, ethyl, hydroxyethyl, isopropyl, cyclopropyl, fluorinated cyclopropyl, cyclobutyl, or fluorinated cyclobutyl; R2is selected from fluorine or methyl; R3is selected from hydroxyl, amino, or methyl; R4is selected from methyl or deuterated methyl.

3. The compound of claim 1 or 2, wherein: Structural unit selected from the group consisting of: or Preferably, the structural unit selected from the group consisting of: or 4. The compound according to any one of claims 1 to 3, characterized in that: The compound is selected from: or 5. A pharmaceutical composition characterized by: which is a compound of any one of claims 1-4 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitroso, isotopically-labeled, metabolite, or prodrug thereof as an active ingredient, in combination with a pharmaceutically acceptable carrier.

6. Use of a compound of any one of claims 1-4 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitroso, isotopically-labeled, metabolite, or prodrug thereof and the pharmaceutical composition of claim 5 in the manufacture of an NLRP3 inhibitor.

7. Use of a compound of any one of claims 1-4 or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, nitroso, isotopically-labeled, metabolite, or prodrug thereof and the pharmaceutical composition of claim 5 in the manufacture of a medicament for preventing and / or treating an NLRP3-related disease.

8. Use according to claim 7, characterized in that: The NLRP3-related disease includes an inflammatory disease, an autoimmune disease, a cardiovascular system disease, a cancer, a renal system disease, a gastrointestinal disease, a respiratory system disease, an endocrine system disease, or a central nervous system disease.

9. Use according to claim 7, characterized in that: The NLRP3-related disease includes cryopyrin-associated periodic syndromes, Muckle-Wells syndrome, familial cold autoinflammatory syndrome, neonatal-onset multisystem inflammatory disease, familial Mediterranean fever, nonalcoholic steatohepatitis, alcoholic liver disease, graft-versus-host disease, multiple sclerosis, rheumatoid arthritis, type I / II diabetes and related complications, psoriasis, Alzheimer's disease, atherosclerosis, gout, chronic kidney disease, sepsis, liver fibrosis, idiopathic pulmonary fibrosis, epilepsy, neuropathic pain, depression, Parkinson's disease, asthma, acute myocardial infarction, lupus erythematosus, rheumatoid arthritis, Crohn's disease, ulcerative colitis, inflammatory bowel disease, rheumatoid arthritis, multiple sclerosis, bronchial asthma, acute respiratory distress syndrome, chronic obstructive pulmonary disease, or ischemic stroke.

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