KR20220039408A - Nlrp3 inflammasome inhibitor and uses thereof - Google Patents

Abstract

The present invention relates to an NLRP3 inflammasome inhibitor and uses thereof for the treatment of NLRP3 inflammasome-mediated inflammatory diseases. More particularly, the present invention provides: an NLRP3 inflammasome inhibitor comprising a compound having an excellent inflammasome inhibitory effect or a pharmaceutically or food acceptable salt thereof; and a pharmaceutical composition and a health functional food composition for preventing or treating NLRP3 inflammasome-mediated inflammatory diseases. The compound or the salt thereof significantly inhibits NLRP3 inflammasome activity, thereby being able to effectively prevent, alleviate or treat various diseases according to the activation of NLRP3 inflammasome.

Description

NLRP3 inflammasome inhibitors and uses thereof

The present invention relates to NLRP3 inflammasome inhibitors and their use for the treatment of NLRP3 inflammasome mediated inflammatory diseases.

Inflammasome is a large multiprotein complex that plays an important role in innate immunity involved in the production of proinflammatory cytokines interleukin-1β (IL-1β) and IL-18. The cytokines involved induce a variety of biological effects related to infection, inflammatory and autoimmune processes. They are inactive precursors, produced from pro-IL-1β and pro-IL-18, and share a common maturation mechanism that requires caspase-1. Caspase-1 itself is synthesized as a zymogen of pro-caspase-1, and produces two subunits that form active caspase-1 through an autocatalytic process. . The activity of caspase-1 occurs within the inflammasome after its assembly.

The most characteristic inflammasome is the NLRP3 (NOD-like receptor pyrin domain-containing protein 3, or NALP3) inflammasome, also called cryopyrin, in response to infection and cell damage, including IL-1β, IL- 18 and induces pyroptosis.

However, the function of the NLRP3 inflammasome can harm the human body, and abnormal or chronic activation of the NLRP3 inflammasome is associated with pathologies such as type 2 diabetes, gouty arthritis, cardiovascular disease, Alzheimer's disease, and cryopyrin-associated- periodic-syndrome), as it is associated with rare genetic disorders. Recently, related research has been rapidly progressing to understand the activation of NLRP3 inflammasome and to develop new therapeutic agents.

Republic of Korea Patent Publication No. 10-2016-0009667 (published on January 26, 2016)

It is an object of the present invention to provide an NLRP3 inflammasome inhibitor comprising a compound having an excellent inflammasome inhibitory effect.

Another object of the present invention is to provide a composition for preventing or treating NLRP3 inflammasome-mediated inflammatory disease comprising the compound.

Another object of the present invention is to provide a reagent composition for inhibiting NLRP3 inflammasome activity comprising the compound.

Another object of the present invention is to provide a method for inhibiting NLRP3 inflammasome activity comprising the step of treating the compound.

In order to achieve the above object, the present invention provides an NLRP3 inflammasome inhibitor comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating NLRP3 inflammasome-mediated inflammatory disease, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention provides a health functional food composition for preventing or improving NLRP3 inflammasome-mediated inflammatory disease, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention provides a reagent composition for inhibiting NLRP3 inflammasome activity containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a method for inhibiting NLRP3 inflammasome activity, comprising treating an animal other than a human with a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

<Formula 1>

The compound according to the present invention or a pharmaceutically or pharmaceutically acceptable salt thereof significantly inhibits the activity of NLRP3 inflammasome induced by urate (MSU) crystals, ATP, nigericin, etc., The compound or a salt thereof can be used as an NLRP3 inflammasome inhibitor, a composition for preventing, improving, or treating NLRP3 inflammasome-mediated inflammatory diseases, etc. Or it can be treated.

1 is an LC-MS analysis of the compound used in Experimental Example of the present invention.
2 and 3 confirm the inhibitory effect of NLRP3 inflammasome activity induced by urate (MSU) crystals according to Experimental Example 1 of the present invention.
4 to 6 confirm the inhibitory effect of NLRP3 inflammasome activity induced by ATP (adenosine triphosphate) or nigericin according to Experimental Example 2 of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides an NLRP3 inflammasome inhibitor comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

<Formula 1>

The compound or its salt may be separated or extracted by a method well known in the art, or may be prepared by chemical synthesis, and a commercially available one may be selected and used, but the method or material is not particularly limited.

Preferably, the compound is 1-(acetyloxy)-4-(1H-indol-3-yl)naphthalen-2-yl acetate [ 1-(acetyloxy)-4-(1H-indol-3-yl)naphthalen -2-yl acetate ].

The compound may be used in the form of a pharmaceutically or food-acceptable salt within the range having the same efficacy.

As used herein, "pharmaceutically or food acceptable" means a salt having a safety and efficacy profile suitable for administration to humans without toxicity to cells or humans exposed to the composition.

The salt may be used in the form of any one of pharmaceutically or food-acceptable basic salt or acid salt. The basic salt can be used in the form of any one of an organic basic salt and an inorganic basic salt, and includes a sodium salt, a potassium salt, a calcium salt, a lithium salt, a magnesium salt, a cesium salt, an aminium salt, an ammonium salt, a triethylamine salt, and pyrithyl. It may be selected from the group consisting of dinium salts.

As the acid salt, an acid addition salt formed by a free acid is useful. As the free acid, an inorganic acid and an organic acid can be used. As the inorganic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfurous acid, phosphoric acid, double phosphoric acid, nitric acid, etc. can be used. As the organic acid, citric acid, acetic acid, maleic acid, malic acid, and fumaric acid can be used. , glucoic acid, methanesulfonic acid, benzenesulfonic acid, camphorsulfonic acid, oxalic acid, malonic acid, glutaric acid, acetic acid, glycolic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, Glutamic acid, citric acid, aspartic acid, stearic acid, etc. may be used, but is not limited thereto, and salts formed using various inorganic and organic acids commonly used in the art may be included.

In addition, the compound may include all salts, hydrates, solvates, derivatives, etc. that can be prepared by a conventional method as well as the salt. The addition salt can be prepared by a conventional method, and is dissolved in a water-miscible organic solvent, for example, acetone, methanol, ethanol, or acetonitrile, and an excess of an organic base is added or an aqueous base of an inorganic base is added, followed by precipitation or crystallization. It can be manufactured by Alternatively, an addition salt may be obtained by evaporating the solvent or excess base from the mixture and drying, or it may be prepared by suction filtration of the precipitated salt.

In the NLRP3 inflammasome inhibitor according to the present invention, the compound or a salt thereof can inhibit the activity of the NLRP3 inflammasome.

Inhibition of the activity of the NLRP3 inflammasome means an in vivo modification that causes a decrease in inflammasome activation of a target cell, etc., and for the purpose of the present invention, inhibition of inflammasome production or cytokine caused by inflammasome It may be production inhibition, but is not limited thereto.

More specifically, the compound or a salt thereof is NLRP3 inflammasome induced by at least one selected from the group consisting of monosodium urate monohydrate (MSU) crystals, adenosine triphosphate (ATP) and nigericin. can inhibit the activity of

In addition to this, the inflammasome may be activated by pathogen components, exogenous factors such as UV irradiation, cholesterol crystals, amyloid β, saturated fatty acids, aluminum hydroxide, etc., but is not limited thereto. .

In addition, the compound or a salt thereof may inhibit the production of interleukin-1β (IL-1β), IL-18 or caspase-1.

More specifically, the compound or a salt thereof may inhibit the production of IL-1β, IL-18 or caspase-1 activated by at least one selected from the group consisting of MSU, ATP, and nigericin.

According to an experimental example of the present invention, it can be confirmed that the production of IL-1β or caspase-1 is significantly inhibited when the compound is treated in cells in which NLRP3 inflammasome is activated by MSU, ATP or nigericin. there was.

The present invention provides a pharmaceutical composition for preventing or treating NLRP3 inflammasome-mediated inflammatory disease, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

<Formula 1>

The compound or a salt thereof may inhibit NLRP3 inflammasome activity.

More specifically, the compound or a salt thereof may inhibit the activity of NLRP3 inflammasome induced by one or more selected from the group consisting of MSU, ATP, and nigericin.

The compound or a salt thereof may prevent or treat NLRP3 inflammasome-mediated inflammatory diseases by inhibiting NLRP3 inflammasome activity.

In the pharmaceutical composition according to the present invention, the NLRP3 inflammasome-mediated inflammatory disease is an abnormal disease that occurs when the inflammasome is abnormally and excessively activated, including gout, arthritis, gouty arthritis, atherosclerosis, Alzheimer's, It may be selected from the group consisting of atopic dermatitis, nonalcoholic fatty liver, type 2 diabetes, metabolic syndrome, metabolic disorders and NLRP3-mediated auto-inflammatory disease, and the NLRP3-mediated auto-inflammatory disease is cryopyrin-related periodic syndrome (Cryopyrin). -associated periodic syndromes (CAPS), familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), neonatal-onset multisystem inflammatory disease (NOMID) ), familial Mediterranean fever (FMF), Pyogenic arthritis with pyoderma gangrenosum and acne (PAPA) syndrome, and hyperimmunoglobulinemia with periodic fever D Syndrome (Hyperimmunoglobulinemia D with periodic fever syndrome, HIDS) may be selected from the group consisting of, but is not limited thereto, and may include all related diseases well known in the art.

In the pharmaceutical composition according to the present invention, the pharmaceutical composition may contain 0.01 to 90 parts by weight of the compound or a salt thereof based on 100 parts by weight of the total pharmaceutical composition, but is not limited thereto.

The pharmaceutical composition according to the present invention may be prepared according to a conventional method in the pharmaceutical field. The pharmaceutical composition may be combined with a suitable pharmaceutically acceptable carrier according to the formulation, and if necessary, excipients, diluents, dispersants, emulsifiers, buffers, stabilizers, binders, disintegrants, solvents, etc. may be prepared further comprising there is. The appropriate carrier and the like do not inhibit the activity and properties of the compound according to the present invention or a pharmaceutically acceptable salt thereof, and may be selected differently depending on the dosage form and formulation.

The pharmaceutical composition according to the present invention can be applied in any dosage form, and more specifically, it can be used by formulating oral dosage forms, external preparations, suppositories, and parenteral dosage forms of sterile injection solutions according to conventional methods.

The solid dosage form among the oral dosage forms is in the form of tablets, pills, powders, granules, capsules, etc., and at least one or more excipients, for example, starch, calcium carbonate, sucrose, lactose, sorbitol, mannitol, cellulose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may be included. In addition, the capsule formulation may further include a liquid carrier such as fatty oil in addition to the above-mentioned substances.

Among the oral dosage forms, liquid formulations include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients, for example, wetting agents, sweeteners, fragrances, preservatives, etc. may be included. there is.

The parenteral formulation may include a sterile aqueous solution, a non-aqueous solution, a suspension, an emulsion, a freeze-dried formulation, and a suppository. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, Tween 61, cacao butter, laurin fat, glycerogelatin, and the like can be used. It is not limited thereto, and any suitable agent known in the art may be used.

In addition, the pharmaceutical composition according to the present invention may further add calcium or vitamins to enhance therapeutic efficacy.

In the pharmaceutical composition according to the present invention, the pharmaceutical composition may be administered in a pharmaceutically effective amount.

As used herein, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not to cause side effects.

The effective dose level of the pharmaceutical composition may be determined according to the purpose of use, the age, sex, weight and health status of the patient, the type of disease, the severity, the activity of the drug, the sensitivity to the drug, the administration method, the administration time, the administration route and the excretion rate, the treatment The duration, formulation, or concomitant use may be determined differently depending on factors including drugs and other factors well known in the medical field. For example, although not constant, generally 0.001 to 100 mg/kg, preferably 0.01 to 10 mg/kg, may be administered once to several times a day. The above dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition according to the present invention may be administered to any animal capable of developing NLRP3 inflammasome-mediated inflammatory disease, and the animal may be, for example, not only humans and primates, but also cattle, pigs, horses, and dogs such as livestock, etc. may include

The pharmaceutical composition according to the present invention may be administered by an appropriate administration route according to the form of the formulation, and may be administered via various routes, either oral or parenteral, as long as it can reach the target tissue. The administration method is not particularly limited, and for example, oral, rectal or intravenous, muscle, skin application, respiratory inhalation, intrauterine dural or intracerebroventricular injection, etc. can be administered in a conventional manner. there is.

The pharmaceutical composition according to the present invention may be used alone for the prevention or treatment of NLRP3 inflammasome-mediated inflammatory disease, or may be used in combination with surgery or other drug treatment.

The present invention provides a health functional food composition for preventing or improving NLRP3 inflammasome-mediated inflammatory disease, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

<Formula 1>

The compound or a salt thereof may inhibit NLRP3 inflammasome activity.

More specifically, the compound or a salt thereof may inhibit the activity of NLRP3 inflammasome induced by one or more selected from the group consisting of MSU, ATP, and nigericin.

The compound or a salt thereof may prevent or ameliorate NLRP3 inflammasome-mediated inflammatory diseases by inhibiting NLRP3 inflammasome activity.

In the health functional food composition according to the present invention, the NLRP3 inflammasome-mediated inflammatory disease is an abnormal disease that occurs when the inflammasome is abnormally and excessively activated, including gout, arthritis, gouty arthritis, atherosclerosis, It may be selected from the group consisting of Alzheimer's disease, atopic dermatitis, nonalcoholic fatty liver, type 2 diabetes, metabolic syndrome, metabolic disorders, and NLRP3-mediated auto-inflammatory diseases, and the NLRP3-mediated auto-inflammatory disease is cryopyrin-related periodic syndrome. (Cryopyrin-associated periodic syndromes, CAPS), familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), neonatal-onset multisystem inflammatory disease , NOMID), familial Mediterranean fever (FMF), Pyogenic arthritis with pyoderma gangrenosum and acne (PAPA) syndrome, and hyperimmunoglobulin with periodic fever It may be selected from the group consisting of Hyperimmunoglobulinemia D with periodic fever syndrome (HIDS), but is not limited thereto, and may include all related diseases well known in the art.

In the health functional food composition according to the present invention, the health functional food composition may contain 0.01 to 90 parts by weight of the compound or its salt based on 100 parts by weight of the total health functional food composition, but is not limited thereto.

In the health functional food composition according to the present invention, the health functional food may be prepared as powder, granule, tablet, capsule, syrup or beverage for the purpose of preventing or improving NLRP3 inflammasome-mediated inflammatory disease. There is no limitation in the form that the health functional food can take, and it can be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods.

In the health functional food composition according to the present invention, the health functional food may include all foods in a conventional sense. For example, beverages and various drinks, fruits and their processed foods (canned fruit, jam, etc.), fish, meat and their processed foods (ham, bacon, etc.), breads and noodles, cookies and snacks, dairy products (butter, cheese, etc.) ), etc. are possible, and may include all functional foods in a conventional sense. It may also include food used as feed for animals.

The health functional food composition according to the present invention may be prepared by further including pharmaceutically acceptable food additives (food additives) and other suitable auxiliary ingredients commonly used in the art. Whether or not it is suitable as a food additive can be judged according to the standards and standards for the relevant item in accordance with the general rules and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety, unless otherwise specified. The items listed in the 'Food Additives Codex' include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high pigment, and guar gum; Mixed preparations, such as a sodium L-glutamate preparation, a noodle-added alkali agent, a preservative preparation, and a tar color preparation, etc. are mentioned.

The other auxiliary ingredients include, for example, flavoring agents, natural carbohydrates, sweeteners, vitamins, electrolytes, coloring agents, pectic acid, alginic acid, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents, etc. may further contain. In particular, as the natural carbohydrate, monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol can be used. , As the sweetener, natural sweeteners such as taumatine and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used.

The effective dose of the compound or its salt contained in the health functional food according to the present invention may be appropriately adjusted according to the purpose of use, such as prevention or improvement of NLRP3 inflamasome-mediated inflammatory disease.

The health functional food composition uses food as a raw material and has the advantage of not having side effects that may occur when taking general drugs for a long time, and has excellent portability, so it can be taken as an adjuvant for preventing or improving NLRP3 inflammasome-mediated inflammatory disease. there is.

In addition, the present invention provides a reagent composition for inhibiting NLRP3 inflammasome activity containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

<Formula 1>

In addition, the present invention provides a method for inhibiting NLRP3 inflammasome activity, comprising treating an animal other than a human with a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

<Formula 1>

Corresponding features may be substituted for the above-mentioned parts.

Hereinafter, examples will be described in detail to help the understanding of the present invention. However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Example 1> Compound analysis

1 is an LC-MS analysis of the compound used in Experimental Examples of the present invention.

The compound was obtained from the Korea Chemical Research Institute, Korea Compound Bank, and the molecular weight (MW=360.2) corresponding to the compound was confirmed in peck 8 of FIG. 1 (purity 92%).

<Experimental Example 1> NLRP3 induced by urate crystals Confirmation of inhibition of inflammasome activity

1. Experimental method

After separating the bone marrow from C57BL/6 mice according to a known method [ Journal of immunology (Baltimore, Md.:1950) 186, 499-507, 2011], bone marrow-derived primary macrophages (Mouse Bone-marrow-derived macrophage) ) was prepared. The macrophages were cultured in DMEM medium containing 10% (v/v) fetal bovine serum (invitrogen, Carlsbad, CA), penicillin at 10,000 units/ml, and streptomycin at 10,000 mg/ml (Invitrogen).

The cultured BMDM was dispensed on a 96-well plate at a density of 5×10 ⁴ cells/ml and 100 ng/ml of LPS [Purified LPS from Escherichia coli , List Biological Laboratory Inc. (Cambell, CA) purchased, used by dissolving in non-toxic water] was treated for 4 hours. In order to exclude the effect of LPS on the compound, the compound was added after washing the LPS with PBS. Cells were treated with the compounds 0.1 μM, 0.5 μM, and 1 μM for 1 hour, and stimulated with monosodium urate (MSU; Invivogen, 500 μg/ml) crystals, an inflammasome activator, in the medium. The supernatant of the cells was checked using an ELISA kit (R&D systems, Minneapolis, MN) to determine the concentration of IL-1β as an indicator of inflammasome activation. The concentration of the standard curve was 11.72-3000 pg/ml.

2. Experimental results

After inducing NLRP3 inflammasome by treating mouse bone-marrow-derived macrophage with urate (MSU) crystals, 0.1 μM, 0.5 μM, and 1 μM of the compound were treated each. As a result, as shown in FIG. 2 It was also shown that the production of IL-1β was inhibited in a concentration-dependent manner.

In addition, as a result of treatment with 1 μM, 5 μM, and 10 μM of the compound after inducing NLRP3 inflammasome by treatment with MSU, it was shown that the production of IL-1β was inhibited in a concentration-dependent manner as shown in FIG. 3 .

<Experimental Example 2> Confirmation of inhibition of NLRP3 inflammasome activity induced by ATP or nigericin

2-1. Experimental methods and results

THP-1 cells, which are human monocytic cells, contain 10% (v/v) fetal bovine serum (invitrogen, Carlsbad, CA), penicillin at 10,000 units/ml, and streptomycin at 10,000 mg/ml (Invitrogen). was cultured in DMEM medium containing Cultured THP-1 cells were seeded on a 96-well plate at a density of 2×10 ⁴ cells/ml, and 100 nM of PMA [purchased from (Sigma-Aldrich), used by dissolving in DMSO] was treated for 3 days. In order to exclude the effect of PMA on the compound, the compound was added after washing with PBS. Cells were treated with the above compounds 0.1 μM, 0.5 μM, and 1 μM for 1 hour, and the medium was stimulated with ATP (5 mM), an inflammasome activator. Alternatively, cells were treated with the compounds 0.01 μM, 0.1 μM, and 1 μM for 1 hour, and the medium was stimulated with nigericin (10 μM), an inflammasome activator. The supernatant of the cells was checked using an ELISA kit (R&D systems, Minneapolis, MN) to determine the concentration of IL-1β as an indicator of inflammasome activation. The concentration of the standard curve was 0.078-20000 pg/ml.

After inducing NLRP3 inflammasome by treating THP-1 cells with ATP, the compound was treated at 0.1 μM, 0.5 μM, and 1 μM, and as a result, the production of IL-1β was inhibited in a concentration-dependent manner as shown in FIG. 4 .

In addition, after inducing NLRP3 inflammasome by treating the THP-1 cells with nigericin, the compound was treated with 0.01 μM, 0.1 μM, and 1 μM each. As a result, as shown in FIG. 5, the production of IL-1β appeared to be significantly suppressed.

2-2. Experimental methods and results

After separating the bone marrow from C57BL/6 mice according to a known method [ Journal of immunology (Baltimore, Md.:1950) 186, 499-507, 2011], bone marrow-derived primary macrophages (Mouse Bone-marrow-derived macrophage) ) was prepared. The macrophages were cultured in DMEM medium containing 10% (v/v) fetal bovine serum (invitrogen, Carlsbad, CA), penicillin at 10,000 units/ml, and streptomycin at 10,000 mg/ml (Invitrogen).

The cultured BMDM was dispensed on a 6-well plate at a density of 2×10 ⁶ cells/ml and 100 ng/ml of LPS [Purified LPS from Escherichia coli , List Biological Laboratory Inc. (Cambell, CA) purchased, used by dissolving in non-toxic water] was treated for 4 hours. In order to exclude the effect of LPS on the compound, the compound was added after washing the LPS with PBS. Cells were treated with the compounds 0.1 μM, 0.5 μM, and 1 μM for 1 hour, and stimulated with ATP (5 mM), an inflammasome activator, in serum-free medium. The cells were cultured in RIPA buffer (50 mM Tris-HCl, pH7.4, 1% NP-40, 0.25% sodium deoxycholate, 150 mM NaCl, 1 mM EGTA, 1 mM PMSF, 1 mM Na ₃ VO ₄ , 10 μg/ml aprotinin and 10 The eluate eluted with lupeptin μg/ml) was subjected to immunoblot analysis.

At this time, as indicators of inflammasome activation, cleavage of pro-IL-1β to IL-1β and cleavage of pro-Caspase-1 into Caspase-1 were confirmed in the supernatant. At this time, the supernatant was precipitated by adding 1 volume of methanol and 0.25 volume of chloroform, followed by centrifugation at 20,000 g for 10 minutes. The upper layer was removed and the mixture obtained by adding 1 volume of methanol was centrifuged at 20,000 g for 10 minutes to obtain a protein precipitate, dried at room temperature, and then dried in Laemmli buffer (1M Tris-HCl, pH 6.8, 100% glycerol, 10% SDS). , 10% 2-mercaptoethanol and 1% bromophenol blue). These samples were transferred to SDS-PAGE and then electrophoresed on a polyvinylidene difluoride membrane. The foil was probed with a primary antibody and a related HRP-conjugated secondary antibody. Each band was visualized using the ECL system.

After inducing NLRP3 inflammasome by treating mouse bone marrow-derived macrophages with ATP, the compound was treated with 0.1 μM, 0.5 μM, and 1 μM, as shown in FIG. 6 , the production of IL-1β and Caspase-1 was significantly inhibited. appeared to be

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. Do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (10)

An NLRP3 inflammasome inhibitor comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
<Formula 1>

The method of claim 1,
The compound or a salt thereof is
NLRP3 influenza, characterized in that it inhibits the activity of NLRP3 inflammasome induced by at least one selected from the group consisting of urate (monosodium urate monohydrate; MSU) crystal, ATP (adenosine triphosphate) and nigericin llamasome inhibitors. The method of claim 1,
The compound or a salt thereof is
An NLRP3 inflammasome inhibitor, characterized in that it inhibits the production of interleukin-1β (IL-1β) or caspase-1. A pharmaceutical composition for preventing or treating NLRP3 inflammasome-mediated inflammatory disease comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
<Formula 1>

5. The method of claim 4,
The compound or a salt thereof is
A pharmaceutical composition for preventing or treating NLRP3 inflammasome-mediated inflammatory disease, characterized in that it inhibits NLRP3 inflammasome activity. 5. The method of claim 4,
The NLRP3 inflammasome-mediated inflammatory disease,
Cryopyrin-associated periodic syndromes (CAPS), familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), neonatal multiple inflammatory disease (Neonatal) -onset multisystem inflammatory disease (NOMID), familial Mediterranean fever (FMF), pyogenic arthritis with pyoderma gangrenosum and acne (PAPA) syndrome, and periodic fever NLRP3-mediated autoinflammatory disease selected from the group consisting of Hyperimmunoglobulinemia D with periodic fever syndrome (HIDS); Prevention of NLRP3 inflamasome-mediated inflammatory disease, characterized in that the disease is selected from the group consisting of gout, arthritis, gouty arthritis, atherosclerosis, Alzheimer's, atopic dermatitis, nonalcoholic fatty liver, type 2 diabetes, metabolic syndrome and metabolic disorders or a therapeutic pharmaceutical composition. 5. The method of claim 4,
The pharmaceutical composition is
Based on 100 parts by weight of the total pharmaceutical composition, a pharmaceutical composition for preventing or treating NLRP3 inflammasome-mediated inflammatory disease, characterized in that the compound or its salt is contained in an amount of 0.01 to 90 parts by weight. A health functional food composition for preventing or improving NLRP3 inflammasome-mediated inflammatory disease comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
<Formula 1>

A reagent composition for inhibiting NLRP3 inflammasome activity comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.
<Formula 1>

A method for inhibiting NLRP3 inflammasome activity, comprising treating an animal other than a human with a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
<Formula 1>

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