Composition for alleviating or treating atopic dermatitis comprising sesamine

Abstract

The present invention relates to a composition for preventing, alleviating or treating atopic dermatitis, comprising sesamine. According to the present invention, when applied to T cells stimulated by an atopic dermatitis-inducing factor, sesamine induces apoptosis thereof, reduces the expression of IL-2 mRNA and CD69, suppresses the proliferation of the T cells, physically binds to and inhibits MCL-1, reduces heterodimerization between MCL-1 and Bak. When administered to an animal model in which actual atopic dermatitis was induced, sesamine exhibits an excellent therapeutic effect by increasing a T cell-mediated immune response, and thus can be advantageously used for the prevention and treatment of atopic dermatitis.

Description

Composition for improving or treating atopic dermatitis containing sesamin

[0001] The present invention relates to a composition for improving or treating atopic dermatitis, comprising sesamin.

[0002] Atopic dermatitis (AD) is a skin disorder characterized by intense itching of unknown etiology, along with dryness, erythema, and inflammation. Atopic dermatitis is caused by a multitude of factors, including immunological abnormalities such as T lymphocyte activation, cytokine dysfunction, decreased cell-mediated immunity, and increased IgE levels, as well as physiological factors and biochemical defects in the skin. The pathogenesis of acute AD has been reported to be related to a Th2 inflammatory response mediated by dermal infiltration of CD4+ T cells and eosinophils, and increased secretion of immunoglobulin E (IgE) and Th2 cytokines. Atopic dermatitis is accompanied by symptoms such as severe pruritus and dry skin, while atopy is characterized by high blood IgE levels and increased eosinophils.

[0003] Atopic dermatitis (AD) is a chronic, relapsing, non-infectious dermatitis resulting from an inflammatory response, leading to various skin disorders. Atopic dermatitis is caused by skin hypersensitivity and inflammation, leading to epithelial dysfunction and immunoallergic responses that impact the skin's unique immune microenvironment. Immune system dysfunction, particularly in two major CD4+ T cell subsets, is a pathologically significant feature of atopic dermatitis. Atopic dermatitis, driven by this systemic dysfunction, is determined by a Th2-dominated immune response, induced by immune cell influx into the skin, serum IgE, and peripheral eosinophilia. Furthermore, atopic dermatitis inflammation induces an increase in Th2 cells in acute skin lesions and the overexpression of Th2-type cytokines, such as IL-2, IL-5, and IL-13. However, in chronic atopic dermatitis, Th1-type cytokines such as IFN-γ are expressed, and an influx of immune dendritic epithelial cells, macrophages, and eosinophils that produce IL-12 occurs.

[0004] Although extensive research has been conducted on the causes and treatments of atopic dermatitis, the complexity of the disease itself and conflicting research data have prevented a clear theory of its exact pathogenesis and effective treatment. Therefore, treatments currently available for atopic dermatitis include steroids, topical anti-inflammatory agents such as tacrolimus and pimecrolimus, antihistamines, and immunosuppressants such as cyclosporine. Additionally, hypoallergenic moisturizers and UVA photochemotherapy are being clinically tested as adjunctive therapies. However, these treatments and therapies primarily aim to manage symptoms rather than provide a fundamental cure, failing to fully address the needs of atopic dermatitis patients.

[0005] Furthermore, until recently, topical steroid preparations were the main treatment for atopic dermatitis, but they have various side effects such as Cushing's syndrome with long-term use, and systemic weakness, fever, muscle pain, joint pain, and loss of appetite due to acute adrenal insufficiency when long-term administration is suddenly discontinued. In addition, infections can be worsened due to immune system disorders, and in particular, when applied externally to the skin for a long period of time, not only can the side effect of weakening skin resistance occur, but when use is discontinued, a serious symptom called steroid contact dermatitis can occur. Therefore, there is a problem that a novel treatment for atopic dermatitis that shows excellent therapeutic effects without these side effects is needed.

[0006] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating atopic dermatitis.

[0007] In addition, another object of the present invention is to provide an over-the-counter drug for preventing or improving atopic dermatitis.

[0008] In addition, another object of the present invention is to provide a cosmetic composition for preventing or improving atopic dermatitis.

[0009] In addition, another object of the present invention is to provide a food composition for preventing or improving atopic dermatitis.

[0010] In addition, another object of the present invention is to provide a health functional food composition for preventing or improving atopic dermatitis.

[0011] In addition, another object of the present invention is to provide a method for preventing or treating atopic dermatitis.

[0012] In order to achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0013] In addition, in order to achieve the other purpose mentioned above, the present invention provides an over-the-counter drug for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0014] In addition, in order to achieve the above-mentioned other object, the present invention provides a cosmetic composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0015] In addition, in order to achieve the above-mentioned other object, the present invention provides a food composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0016] In addition, in order to achieve the above-mentioned other object, the present invention provides a health functional food composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0017] In addition, in order to achieve the above-mentioned another object, the present invention provides a method for preventing or treating atopic dermatitis, comprising a step of administering sesamin or a pharmaceutically acceptable salt thereof to a subject.

[0018] According to the present invention, when sesamin was treated to T cells stimulated with an atopic dermatitis-inducing factor, it induced cell death thereof, reduced IL-2 mRNA and CD69 expression, inhibited the proliferation of the T cells, physically bound to the MCL-1 active site and inhibited it, reduced heterodimerization between MCL-1 and Bak, and when treated to an animal model in which actual atopic dermatitis was induced, it increased the T cell-mediated immune response, showing an excellent therapeutic effect, and thus can be usefully utilized for the prevention and treatment of atopic dermatitis.

[0019] Figure 1 shows the results of confirming the IL-2 production, CD69 expression, and proliferation inhibition effects of sesamin in activated T cells.

[0020] Figure 2 shows the results confirming that IL-2 production is suppressed when activated T cells are pretreated with sesamin.

[0021] Figure 3 shows the results of confirming the physical interaction of sesamin with MCL-1, a predicted target molecule, in activated T cells.

[0022] Figure 4 shows the results of molecular docking to confirm the physical interaction of sesamin with MCL-1, a predicted target molecule, in activated T cells.

[0023] Figure 5 shows the prediction results and molecular docking analysis results of the top three candidate proteins predicted to bind to sesamin.

[0024] Figure 6 shows the results confirming that when activated T cells were treated with sesamin, MCL-1 activity was suppressed, thereby regulating T cell activation.

[0025] Figure 7 shows the results confirming that inhibition of MCL-1 activity by sesamin blocks heterodimerization of MCL-1 and Bak in activated T cells.

[0026] Figure 8 shows the results confirming that sesamin selectively induces the cell death pathway only in activated T cells.

[0027] Figure 9 shows the results confirming that sesamin enhances the apoptotic population only in activated T cells.

[0028] Figure 10 shows the results confirming that sesamin promotes the cleavage of caspase-3 and caspase-8 in activated T cells.

[0029] Figure 11 shows the results confirming that symptoms improved when sesamin was orally administered to an animal model induced with atopic dermatitis.

[0030] Figure 12 shows the results confirming that pathological manifestations were improved when sesamin was orally administered to an animal model induced with atopic dermatitis.

[0031] Figure 13 shows the results confirming that when sesamin was orally administered to an animal model that induced atopic dermatitis, the expression of genes related to atopic dermatitis was reduced in the ear tissue of the animal model.

[0032] Figure 14 shows the results confirming that oral administration of sesamin to an animal model induced with atopic dermatitis did not affect the mRNA expression of IL-6 and TSLP.

[0033] Figure 15 shows the results confirming that when sesamin was orally administered to an animal model induced with atopic dermatitis, the systemic immune response was improved.

[0034] Figure 16 shows the results of confirming that when sesamin is orally administered to an animal model that has induced atopic dermatitis, it promotes the cell death mechanism by regulating MCL-1 activity.

[0035] Hereinafter, the present invention will be described in more detail.

[0036] The present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0037] In the present invention, the sesamin is characterized in that it is a compound represented by the following chemical formula 1.

[0038]

[0039]

[0040] Sesamin, a fat-soluble lignan found in sesame seeds, is particularly abundant in white sesame seeds and is known to have excellent antioxidant properties. Sesamin is a regulator of cholesterol and tocopherol metabolism and is known to facilitate the biosynthesis of gamma-linolenic acid. Sesamin has been reported to promote alcohol detoxification in the liver, inhibit lipid peroxide production, and have anti-carcinogenic properties.

[0041] In the present invention, the term "pharmaceutically acceptable salt" means a salt that can be used pharmaceutically among salts that are substances in which cations and anions are bonded by electrostatic attraction, and can typically be a metal salt, a salt with an organic base, a salt with an inorganic acid, a salt with an organic acid, a salt with a basic or acidic amino acid, etc. For example, the metal salt can be an alkali metal salt (sodium salt, potassium salt, etc.), an alkaline earth metal salt (calcium salt, magnesium salt, barium salt, etc.), an aluminum salt, etc.; the salt with an organic base can be a salt with triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N-dibenzylethylenediamine, etc.; the salt with an inorganic acid can be a salt with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc. Salts with organic acids may include salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.; salts with basic amino acids may include salts with arginine, lysine, ornithine, etc.; salts with acidic amino acids may include salts with aspartic acid, glutamic acid, etc., but are not limited thereto.

[0042] According to an embodiment of the present invention, when sesamin was administered to T cells stimulated with an atopic dermatitis-inducing factor, it was confirmed that at least one selected from the group consisting of T cell activation, function, and proliferation was inhibited. Furthermore, sesamin promoted apoptosis of activated T cells.

[0043] In particular, it was confirmed that sesamin physically binds to the MCL-1 active site of activated T cells, inhibiting MCL-1 and thereby inhibiting the formation of heterodimers between MCL-1 and Bak. Furthermore, when treated in an animal model induced with actual atopic dermatitis, it showed excellent therapeutic effects by increasing T cell-mediated immune responses, and thus can be usefully utilized for the prevention and treatment of atopic dermatitis.

[0044] In addition, according to another embodiment of the present invention, when sesamin and UMI-77, a Bcl-2 inhibitor, were co-treated, a better MCL-1 activity inhibition and T cell activation control effect was observed than when sesamin was treated alone.

[0045] Therefore, in the present invention, the pharmaceutical composition may additionally include a Bcl-2 inhibitor.

[0046] The above Bcl-2 inhibitor may be at least one selected from the group consisting of UMI-77, obatoclax, navitoclax, venetoclax, sabutoclax, A-1155463, A-1331852, ABT-737, TW-37, A-1210477, AT101, HA14-1, BAM7, S44563 and pharmaceutically acceptable salts thereof, but is not limited thereto. Preferably, it may be UMI-77, but is not limited thereto.

[0047] The term "prevention" of the present invention means any act of suppressing or delaying the onset of atopic dermatitis by administering a composition according to the present invention.

[0048] The term "treatment" in the present invention means any act of improving or beneficially changing the symptoms of the disease by administering the pharmaceutical composition.

[0049] The "pharmaceutical composition" of the present invention is intended for use in the prevention and / or treatment of atopic dermatitis. For prophylactic use, the pharmaceutical composition of the present invention is administered to a subject suspected of having or at risk of developing a disease, disorder, or condition described herein. That is, it can be administered to a subject at risk of developing atopic dermatitis and related diseases thereof. For therapeutic use, the pharmaceutical composition of the present invention is administered to a subject, such as a patient already suffering from a disease described herein, in an amount sufficient to treat or at least partially arrest the symptoms of a disease, disorder, or condition described herein. An amount effective for such use will depend on the severity and course of the disease, disorder, or condition, previous treatment, the subject's health status and responsiveness to the drug, and the judgment of a physician or veterinarian.

[0050] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment. The effective dosage level can be determined based on factors including the subject's type and severity, age, sex, drug activity, drug sensitivity, administration time, administration route and excretion rate, treatment period, concurrently used drugs, and other factors well known in the medical field. For example, the sesamin can be administered at a dosage of 0.01 to 5,000 mg / kg per day, specifically 10 to 1,000 mg / kg, and the administration can be administered once a day or in several divided doses. The pharmaceutical composition can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. It can be administered singly or in multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that achieves the maximum effect with the minimum amount without causing side effects, and this can be easily determined by those skilled in the art.

[0051] In addition, the pharmaceutical composition may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the intended method, and the dosage may vary depending on the patient's condition and weight, the extent of the disease, the drug form, the route of administration, and the time of administration, but may be appropriately selected by those skilled in the art. In the embodiments of the present invention, oral administration was used, but is not limited thereto.

[0052] The term "administration" in the present invention refers to the act of introducing a composition containing sesamin into a subject using an appropriate method. The term "subject" in the present invention refers to any animal, including rats, mice, and livestock, including humans, that has developed or may develop atopic dermatitis. A specific example may be a mammal, including humans.

[0053] The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, excipient or diluent commonly used in the manufacture of pharmaceutical compositions, and the carrier may comprise a non-naturally occurring carrier. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0054] In addition, the pharmaceutical composition may have any one dosage form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizers, and suppositories, and may be various oral or parenteral dosage forms. When formulated, it may be prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

[0055] Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives.

[0056] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0057] In addition, the present invention provides an over-the-counter drug composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0058] In the present invention, the above "quasi-drug" means a fiber, rubber product or similar product used for the purpose of treating, alleviating, managing or preventing a disease of humans or animals; a product that has a weak effect on the human body or does not directly affect the human body, is not an apparatus or machine and similar product; and a product corresponding to one of the preparations used for sterilization, insecticide and similar purposes for preventing infection. It means a product used for the purpose of diagnosing, treating, alleviating, managing or preventing a disease of humans or animals, excluding products that are not apparatuses, machines or devices; and products used for the purpose of exerting a pharmacological effect on the structure and function of humans or animals, excluding products that are not apparatuses, machines or devices; and also includes external skin preparations and personal hygiene products.

[0059] When the sesamin of the present invention is included in an over-the-counter drug for the purpose of preventing or improving atopic dermatitis, the sesamin may be used as is or in combination with other over-the-counter drug ingredients, and may be used appropriately according to conventional methods. The amount of active ingredients mixed may be appropriately determined depending on the intended use.

[0060] The over-the-counter drug of the present invention is not particularly limited thereto, but may be manufactured and used in the form of, for example, a cream, lotion, aerosol, shampoo, gel, or pack.

[0061] In the case of creams, ointments, shampoos, gels or packs, bases such as white petrolatum, yellow petrolatum, lanolin, bleached beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oils, gelling hydrocarbons, polyethylene glycol, liquid paraffin, squalane, etc.; solvents and solubilizing agents such as oleic acid, isopropyl myristate, glycerin triisooctanoate, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, vegetable oils, etc.; antioxidants such as tocopherol derivatives, L-ascorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, etc.; preservatives such as parahydroxybenzoate esters, etc.; moisturizers such as glycerin, propylene glycol, sodium hyaluronate, etc. Surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, hydroxypropylcellulose, and hydroxypropylmethylcellulose.

[0062] In the case of aerosols, bases such as white petrolatum, yellow petrolatum, lanolin, bleached beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oils, gelling hydrocarbons, polyethylene glycol, liquid paraffin, and squalane used in the preparation of ointments, creams, gels, suspensions, emulsions, solutions, and lotions; solvents and solubilizing agents such as oleic acid, isopropyl myristate, diisopropyl adipate, isopropyl sebacate, glycerin triisooctanoate, crotamiton, diethyl sebacate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, and vegetable oils; antioxidants such as tocopherol derivatives, L-ascorbic acid, dibutylhydroxytoluene, and butylhydroxyanisole; preservatives such as parahydroxybenzoic acid esters; Humectants such as glycerin, propylene glycol, and sodium hyaluronate; surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, hydroxypropylcellulose, and hydroxypropylmethylcellulose; In addition, various stabilizers, buffers, coagulants, suspending agents, emulsifiers, fragrances, preservatives, solubilizers, and other suitable additives can be blended. In addition, stabilizers, preservatives, absorption promoters, pH adjusters, and other suitable additives can be blended as needed.

[0063] For the pharmaceutical product of the present invention, a more specific description is the same as the description for the above pharmaceutical composition.

[0064] In addition, the present invention provides a cosmetic composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0065] The components included in the cosmetic composition of the present invention may include, in addition to sesamin as an active ingredient, components commonly used in cosmetic compositions, and may include, for example, conventional auxiliary agents and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0066] The cosmetic composition of the present invention can be manufactured in any formulation commonly manufactured in the art, and examples thereof include emulsion, cream, toner, pack, foundation, lotion, cosmetic solution, hair cosmetic, etc.

[0067] Specifically, the cosmetic composition of the present invention includes formulations of skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser.

[0068] When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component.

[0069] When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0070] In the case where the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

[0071] When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth may be used as carrier components.

[0072] When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

[0073] In addition to the above-mentioned effective ingredient, the cosmetic composition of the present invention may contain one or more ingredients that help improve skin condition, including ingredients that exhibit the same or similar atopic dermatitis treatment activity. The above ingredients are hyaluronic acid, butylene glycol, glycerin, amino acid, trehalose, kojic acid and its derivatives, arbutin, ascorbic acid and its derivatives, hydroquinone and its derivatives, resorcinol, 2,7-dinitroindazole, adenosine, retinol, retinyl palmitate, polyethoxylated retinamide, yeast, dipeptide, palmitoyl oligopeptide & palmitoyl tripeptide-7, acetyl These include, but are not limited to, acetyl hexapeptide, epidermal growth factor (EGF), or plant extracts such as citrus, rice, licorice, shea butter, aloe vera, coconut, olive, and avocado.

[0074] Additionally, the cosmetic composition of the present invention may be provided in a pack. The pack is preferably prepared in any one formulation selected from among a mask pack, a sleeping pack, a cleansing pack, a wash-off pack, and a peel-off pack, but is not limited thereto.

[0075] In addition, the present invention provides a food composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0076] In addition, the present invention provides a health functional food composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

[0077] The food composition according to the present invention includes all forms, such as functional foods, nutritional supplements, health foods, and food additives. The food composition of the above type may be formulated into any form selected from the group consisting of powders, tablets, capsules, pills, and liquids using conventional methods known in the art, but is not limited thereto. The food composition may be manufactured in various forms using methods known in the art.

[0078] For example, as a health food, the sesamin of the present invention or a pharmaceutically acceptable salt thereof may be granulated, encapsulated, or powdered and consumed, or may be prepared in the form of tea, juice, or drinks for consumption. Furthermore, the sesamin of the present invention or a pharmaceutically acceptable salt thereof may be prepared in the form of a composition by mixing it with a known substance or active ingredient known to have a preventive, improving, or therapeutic effect on atopic dermatitis.

[0079] In addition, functional foods can be produced by adding the sesamin of the present invention or a pharmaceutically acceptable salt thereof to beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meats and their processed foods (e.g., ham, sausage, corned beef, etc.), breads and noodles (e.g., udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, taffy, dairy products (e.g., butter, cheese, etc.), edible vegetable oils, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.).

[0080] The preferred content of the sesamin of the present invention or a pharmaceutically acceptable salt thereof in the food composition of the present invention is not limited thereto, but may be, for example, 0.01 to 80 wt% of the final manufactured food, and preferably 0.01 to 50 wt% of the final manufactured food. If the content is less than 0.00001 wt%, the effect is minimal, and if it exceeds 50 wt%, the increase in effect relative to the amount used is minimal, making it uneconomical.

[0081] In addition, in order to use the sesamin of the present invention or a pharmaceutically acceptable salt thereof in the form of a food additive, it can be manufactured and used in the form of tablets, capsules, powder, granules, liquid, pills, etc.

[0082] When the composition of the present invention is manufactured into a beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, just like conventional beverages. The natural carbohydrates mentioned above may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g, per 100 ml of the composition of the present invention.

[0083] In addition to the above, the composition of the present invention may include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may include fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in combination. The proportion of these additives is not particularly critical, but is typically selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

[0084] In the present invention, “health supplement” or “health functional food” means a food manufactured and processed using raw materials or ingredients with functionality useful to the human body according to the Health Functional Food Act, and “functionality” means that it is consumed for the purpose of obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological action.

[0085] The present invention also provides a method for preventing or treating atopic dermatitis, comprising the step of administering sesamin or a pharmaceutically acceptable salt thereof to a subject.

[0086] In addition, the above-mentioned entity may be additionally administered with a Bcl-2 inhibitor, but is not limited thereto.

[0087] In the present invention, the Bcl-2 inhibitor may be at least one selected from the group consisting of UMI-77, obatoclax, navitoclax, venetoclax, sabutoclax, A-1155463, A-1331852, ABT-737, TW-37, A-1210477, AT101, HA14-1, BAM7, S44563, and pharmaceutically acceptable salts thereof, but is not limited thereto. Preferably, it may be UMI-77, but is not limited thereto.

[0088] The above-mentioned subject is preferably a mammal, including a human, and includes all patients who require treatment for atopic dermatitis, including patients who are being treated, patients who have received treatment, and patients who need to receive treatment, and may also include patients who have undergone surgical operation for the treatment of atopic dermatitis.

[0089]

[0090] Throughout this specification, '%' used to indicate the concentration of a particular substance is (w / w)% for solid / solid, (w / v)% for solid / liquid, and (v / v)% for liquid / liquid, unless otherwise stated.

[0091] The terminology used in this specification is intended to appropriately express preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the customs of the field to which the present invention pertains. Therefore, the definitions of these terms should be determined based on the contents of this specification as a whole. Throughout this specification, when a part is said to "include" a certain component, unless specifically stated otherwise, this does not mean that other components are excluded, but rather that other components may be included.

[0092] Unless otherwise defined, all technical terms used in this invention have the same meaning as commonly understood by those skilled in the art. While preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention. The contents of all publications cited as references herein are incorporated herein by reference.

[0093] Hereinafter, the present invention will be described in detail with reference to the attached drawings, using exemplary embodiments. However, the following exemplary embodiments are provided as illustrative examples of the present invention. If a detailed description of a technology or configuration well known to those skilled in the art is judged to unnecessarily obscure the gist of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the following claims and equivalents interpreted therefrom.

[0094]

[0095] Experimental Example 1. Analysis Method

[0096] 1-1. Cell culture

[0097] Jurkat T cells (KCLB number: 40152, clone E6-1) were purchased from the Korea Cell Line Bank (KCLB, SEOUL, KOREA). Mouse CD4+ T cells were isolated from the lymph nodes and spleens of 8-week-old female BALB / c mice (RRID:IMSR_ORNL:IE-BALB / c) using a MojoSort™ magnetic cell separation system (Biolegend, San Diego, CA, USA). Jurkat T cells were cultured in RPMI medium (Welgene, Gyeongsan, Korea) containing 10% fetal bovine serum (FBS; Welgene, Gyeongsan, Korea), 1X penicillin-streptomycin (P-streptomycin; Welgene Gyeongsan, Korea), and 2 mM L-glutamine (L-glutamine; Welgene Gyeongsan, Korea). Mouse CD4+ cells were cultured in a medium supplemented with 1X sodium pyruvate and 50 μM 2-mercaptoethanol. The cells were cultured at 37°C in a humidified incubator containing 5% CO2 and 95% air.

[0098]

[0099] 1-2. Reagents and antibodies

[0100] Sesamin was purchased from ChemFacs (Wuhan, Chian). Human IL-2 Duoset ELISA kit was purchased from R&D Systems (Minneapolis, MN, USA). Mouse IgE OptEIA ELISA set was purchased from BD biosciences (San Diego, CA, USA). Anti-CD69 antibodies conjugated with FITC and AnnexinV / 7AAD apoptosis assay kit were purchased from BioLegend. Quanti-Max for WST assay kit and ECL reagent for Western blotting detection were purchased from Biomax (Guri, Korea). Dynabeads Protein co-IP (G co-immunoprecipitation) assay kit was purchased from CellTraceTM CFSE cell proliferation kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA). To induce atopic dermatitis, house dust mites (HDM) were purchased from Greer (Lenoir, NC, USA). 2,4-dinitrochlorobenzene (DNCB), Umi-77, phorbol 12-myristate 13-acetate (PMA), and A23187 were purchased from Sigma Chemical Co. (St. Louis, MO, USA). CNBr-activated Sepharose 4B was purchased from GE Healthcare (Chicago, IL, USA). LaboPass Q Master SYBR green was purchased from Cosmo Genetech (Seoul, Korea), and Prime Script RT Master was purchased from Takara (Shiga, Japan).Anti-human CD3 antibodies (Cat# BE0001-2, RRID: AB_1107632), human CD28 antibodies (Cat# BE0248, RRID: AB_2687729), anti-mouse CD3 antibodies (Cat# BE0001-1, RRID: AB_ 1107634), and mouse CD28 antibodies (Cat# BE0015-5, RRID: AB_ 1107628) for T cell stimulation were purchased from BioXcell (West Lebanon, NH, USA). Anti-bcl-2 (Anti-bcl-2 (Cat# sc-7382, RRID: AB_626736)) antibody was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The antibodies were: Anti-caspase3 (Cat# 9662, RRID: AB_331439), Anti-caspase8 (Cat# 9746, RRID: AB_2275120), Anti-β-actin (Cat# 9746, RRID: AB_330288), Anti-MCL-1 (Cat#94296, RRID: AB_2722740), Anti-phosphorylated MCL-1 at Ser64 (Cat#13297, RRID: AB_2798173), Anti-Bak (Cat#12105, RRID: AB_2716685), Anti-Bax (anti-Bax (Cat#2772, RRID: AB_329921)) was purchased from cell signaling Technology (Danvers, MA, USA).

[0101]

[0102] 1-3. Mouse

[0103] Eight-week-old female BALB / c mice were purchased from Samtako and housed under specific pathogen-free (SPF) conditions. All animal care and experimental procedures were approved by the Institutional Animal Care and Use Committee of Daegu Catholic University College of Medicine (Approval Number: DCIAFCR-230823-20-Y).

[0104]

[0105] 1-4. T cell stimulation

[0106] T cells pre-incubated with sesamin (0–40 μM concentration) at 37°C for 1 h were stimulated by treatment with fixed anti-CD3 (10 μg / ml) and soluble anti-CD28 (2 μg / ml) antibodies.

[0107]

[0108] 1-5. ELISA (enzyme-linked immunosorbent assay) analysis

[0109] After stimulating T cells with anti-CD3 / CD28 antibodies for 24 hours under each condition, supernatants were collected for ELISA analysis. IL-2 produced in the supernatants was measured using the DuoSet ELISA kit. To measure IgE (immunoglobulin E) levels, blood was collected from sacrificed mice and centrifuged at 7,000 rpm for 5 minutes to obtain serum. Before performing ELISA, the obtained serum was diluted 1:100 according to the instructions of the OptEIA ELISA kit (BD biosciences, San Diego, CA, USA).

[0110]

[0111] 1-6. Measurement of CD69 expression using flow cytometry

[0112] After stimulating Jurkat T cells with anti-CD3 / CD28 antibodies for 16 hours under each condition, cells were collected and stained with FITC-conjugated anti-CD69 antibody for 30 minutes at 4°C. Fluorescence was measured using a flow cytometer (Beckman Coulter) to detect CD69 expression on the cell surface. Mean fluorescent intensity (MFI) and histogram data were generated using BD flow cytometry software.

[0113]

[0114] 1-7. CFSE proliferation assay

[0115] T cell proliferation activity was measured using a CFSE proliferation kit. Mouse CD4+ T cells pre-stained with 0.5 μM CFSE were pretreated with 40 μM sesamin for 1 hour and then stimulated with anti-CD3 / CD28 antibodies for 72 hours. CFSE fluorescence was confirmed by flow cytometry, and MFI and population were acquired using BD flow cytometry software.

[0116]

[0117] 1-8. Apoptosis analysis using AnnexinV and 7AAD kits

[0118] Apoptosis of Jurkat T cells was detected by a double staining method using AnnexinV conjugated with FITC and 7-AAD. Specifically, cultured Jurkat T cells were resuspended in 100 μl of 1X binding buffer (10 mM HEPES, 150 mM NaCl, 5 mM KCl, 5 mM MgCl2, 1.8 mM CaCl2) containing AnnexinV (20 μg / ml) and 7-AAD (1 μg / ml) and incubated at room temperature for 15 min. Fluorescence was detected by flow cytometry (Beckman Coulter), and contour plot data were generated using BD flow cytometry software. The percentages of AnnexinV+7AAD− (early apoptotic population) and AnnexinV+7AAD+ (late apoptotic population) were analyzed from the contour plot data.

[0119]

[0120] 1-9. Co-immunoprecipitation analysis and Western blotting

[0121] To assess the interaction between MCL-1, Bax, and Bak, a co-IP assay was performed. After harvesting stimulated cells, the cell pellet was dissolved in RIPA buffer at 4°C for 30 minutes to prepare a lysate, which was then centrifuged. Co-IP analysis was performed using the prepared lysate and a co-IP kit.

[0122] Specifically, 20 μl of Dynabeads Protein A was added and incubated at 4°C for 1 hour to conjugate with the immunoprecipitated antibody. The conjugate was incubated with the lysate prepared for ON at 4°C and washed twice with washing buffer. The SDS loading buffer and heat-denatured eluted samples were loaded onto a 10% SDS-PAGE gel for separation, transferred to nitrocellulose membranes, and blocked in 5% skim milk prepared using TBS containing 0.1% Tween 20 (TBS-T) for 1 hour. After washing with TBS-T, the membranes were incubated overnight at 4°C with immunoblotting antibodies in 3% skim milk using TBS-T. Excess primary antibodies were removed by washing the membranes four times with TBS-T. The membranes were then incubated with 0.1 μg / ml peroxidase-labeled secondary antibodies (to rabbit or mouse) at room temperature for 1.5 h. After washing four times with TBS-T, bands were visualized using ECL Western blot detection reagents (Thermo Fisher Scientific, Waltham, MA, USA) with the Syngene G-BOX Chemi-XRQ gel documentation system. All detected bands were quantified by ImageJ and normalized to the intensity of the loading control protein.

[0123]

[0124] 1-10. Pull-down analysis

[0125] To confirm the physical interaction between sesamin and MCL-1, sesamin-Sepharose 4B beads were generated.

[0126] Specifically, activated dry powder of CNBr-activated Sepharose 4B beads dissolved in 1 mM HCl was added to 2 mg of sesamin dissolved in DMSO or DMSO in coupling buffer (0.1 M NaHCO3 (pH 8.3) and 0.5 M NaCl). After mixing overnight, the coupling buffer was removed and replaced with 0.1 M Tris-HCl buffer (pH 8.0). The conjugate was rotated overnight and washed once with 0.1 M acetate buffer containing 0.5 M NaCl (pH 4.0), followed by a second wash with 0.1 M acetate buffer containing 0.5 M NaCl. Jurkat T cell lysates were incubated overnight with control Sepharose 4B beads or sesamin-Sepharose 4B beads (SS-beads) at different ratios (100:0, 50:50, 0:100) in reaction buffer (50 mM Tris, 5 mM EDTA, 150 mM NaCl, 1 mM DTT, 0.01% NP-40, 2 mg / ml BSA, 0.02 mM PMSF, and 1 μg of protease inhibitor). After incubation, the complexes were washed with washing buffer (50 mM Tris, 5 mM EDTA, 150 mM NaCl, 1 mM DTT, 0.01% NP-40, and 0.02 mM PMSF) and eluted with SDS loading buffer. After gel separation, MCL-1, Bak, and Bax were detected by Western blotting, and the level of precipitated MCL-1 was analyzed through normalization.

[0127]

[0128] 1-11. RNA extraction and real-time quantitative PCR analysis

[0129] Total RNA was isolated from Jurkat T cells, ear tissue, or draining lymph nodes using TRIzol reagent. After quantifying the RNA concentration, complementary DNA was synthesized using 2 μg of total RNA and Prime serript RT Master. Quantitative real-time PCR was then performed using LaboPass Q Master SYBR green according to the manufacturer's instructions. To obtain relative fold values, the expression of the GAPDH (glyceraldehyde 3-phosphate dehydrogenase) gene was normalized using the ΔΔCT method. Gene expression was calculated using the following mathematical equation (1).

[0130] [Mathematical Formula 1]

[0131] ΔΔCT = (CT Target -CT GAPDH ).

[0132] The experiment was repeated at least three times, and the primers in Table 1 below were used for this purpose.

[0133] Gene name Sequence direction Sequence number human IL2 CAC GTC TTG CAC TTG TCA C Forward 1 CCT TCT TGG GCA TGT AAA ACT Reverse 2 human GAPDHC GG AGT CAA CGG ATT TGG TCG TAT Forward 3 AGC CTT CTC CAT GGT GGT GAA GAC Reverse 4 mouse il2 TGA GCA GGA TGG AGA ATT ACA GG Forward 5 GTC CAA GTT CAT CTT CTA GGC AC Reverse 6 mouse il4 ACA GGA GAA GGG ACG CCA T Forward 7 GAA GCC GTA CAG ACG AGC TCA Reverse 8 mouse il6 CCG GAG AGG AGA CTT CAC AG Forward 9 GGA AAT TGG GGT AGG AAG GA Reverse 10 mouse il 13 GCA ACA TCA ACA GGA CCA GA Forward 11 GTC AGG GAA TCC AGG GCT AC reverse 12 mouse il 31 TCG GTC ATC ATA GCA CAT CTG GAG forward 13 GCA CAG TCC CTT TGG AGT TAA GTC reverse 14 mouse ifng TCA AGT GGC ATA GAT GTG GAA GAA forward 15 TGG CTC TGC AGG ATT TTC ATG reverse 16 mouse il 17 TCC CCT CTG TCA TCT GGG AAG forward 17 CTC GAC CCT GAA AGT GAA GG reverse 18 mouse tslpAGG CTA CCC TGA AAC TGA G forward 19 GGA GAT TGC ATG AAG GAA TAC C reverse 20 mouse gapdh GCA CAG TCA AGG CCG AGA AT forward 21 GCC TTC TCC ATG GTG GTG AA reverse 22

[0134]

[0135] 1-12. Cell viability confirmed by the trypan blue dye exclusion method and proliferation analysis by the WST-8 assay.

[0136] Cell viability was determined using the trypan blue dye exclusion method. Specifically, Jurkat T cells cultured under each condition were stained with trypan blue and loaded onto slides for a LUNA automated cell counter (Logos Biosystems, Anyang, Korea). Based on the trypan blue-stained cells, the number of live and dead cells was determined, confirming cell viability.

[0137] Additionally, Jurkat T cells cultured under each condition were stained with 10 μl of WST-8 solution for 20 min. The optical density was then measured at 450 nm using an iMARK microplate reader (Bio-Rad Laboratories, Inc., Berkeley, CA, USA). The proliferation ratio was calculated and expressed as a fold increase compared to the control group.

[0138]

[0139] 1-13. Identifying Related Genes in the Genecards Databank

[0140] To determine the target proteins of sesamin in T cells, immune-related, T-cell function-related, and lignan-related genes were identified from the Genecards databank. Venn diagram analysis was used to sequentially identify overlapping genes. After obtaining 287 lignan-related genes, 20 candidate target proteins based on the structure of sesamin were predicted on the Swisstargetprediction server (http: / / www.swisstargetprediction.ch) using the standard SMILES code (C1C2C(COC2C3=CC4=C(C=C3)OCO4)C(O1)C5=CC6=C(C=C5)OCO6) obtained from PubChem. Probabilities were calculated based on cross-validation. As a result, three candidate proteins, MCL-1, HIF1A, and ALOX5, were identified among the 20 candidates based on probabilities (>0.1).

[0141]

[0142] 1-14. Molecular docking analysis

[0143] Molecular docking analysis was performed using AMDock software to visualize the interactions between target proteins and small molecules. Furthermore, binding energy values, including affinity energy, Ki values, and ligand efficiency, were obtained. The 3D structures of MCL-1, HIF1A, and ALOX5 used in the molecular docking analysis were downloaded from the PDB using accession numbers 5FDR, 4ZPR, and 3O8Y, respectively. Docking poses were displayed using pyMOL software.

[0144]

[0145] 1-15. Preparation of an animal model for atopic dermatitis and treatment with test substances

[0146] First, five mice were randomly divided into five groups: healthy control mice not treated with HDM extract / DNCB or sesamin (Control), control mice treated with 25 mg / kg sesamin alone (SS), control mice treated with HDM extract / DNCB alone (AD), experimental group mice treated with HDM extract / DNCB and 10 mg / kg sesamin simultaneously (AD+SS(10 mg / kg)), experimental group mice treated with HDM extract / DNCB and 25 mg / kg sesamin simultaneously (AD+SS(25 mg / kg)), and control group mice treated with HDM extract / DNCB and 20 mg / kg tofacitinib simultaneously (AD+Tofa (25 mg / kg)).

[0147] To induce atopic dermatitis (AD) in each experimental group, the surface of both earlobes was peeled off four times with surgical tape (Seoil Chemical, Hwaseong, Korea). After peeling, each ear was treated with 10 μl of DNCB (1%), and 4 days later, 10 μl of HDM extract (10 mg / ml). HDM extract / DNCB treatments were repeated weekly for 4 weeks. Treatment was started 1 day after the second DNCB application, and after a 2-day rest, oral sesamin administration was repeated for 4 weeks in a cycle of 5 days of administration and 2 days of rest. Ear thickness was measured 24 hours after application of HDM extract or DNCB using a dial thickness gauge (Kori Seiki MFG Co., Tokyo, Japan). Body weight was assessed on days 0, 14, and 28 after AD induction, and the number of scratches was assessed on day 28. Mice in each experimental group were euthanized on day 28.

[0148]

[0149] 1-16. Histopathological analysis

[0150] After sacrificing mice in each experimental group, ear tissues were collected. The removed ear tissues were fixed in 10% paraformaldehyde and embedded in paraffin. The paraffin-embedded tissues were cut into 5-μm-thick sections, deparaffinized, and stained with hematoxylin and eosin (H&E). The clinical scores for the thickness of the dermis and epidermis and the dermal papilla were measured using H&E-stained slides. The criteria for clinical scoring for inflammation were as follows: no signs of inflammation (0 points); mild inflammatory response, a small number of immune cell infiltrations, and minimal tissue changes (1 point); definite inflammatory response, moderate immune cell infiltration, and moderate tissue changes (2 points); and severe inflammatory response, severe tissue changes with a large number of immune cell infiltrations and severe epidermal hyperplasia (3 points).

[0151] In addition, the criteria for clinical scoring for the dermal papilla are as follows: dilation or elongation of the dermal papilla, mild cellular infiltration, and mild changes observed (1 point); marked dilation and elongation of the dermal papilla, and structural changes observed in the dermis around the nipple (2 points); severe dilation and elongation of the dermal papilla, and massive inflammatory cell infiltration observed with severe structural changes (3 points).

[0152] To determine the number of infiltrated mast cells at this time, the sections were stained with 0.01% toluidine blue, and the number of infiltrated mast cells in randomly selected areas was counted.

[0153]

[0154] 1-17. Statistics

[0155] The mean values ​​for each experiment were calculated from data obtained from three separate in vitro experiments performed on separate days. For in vivo studies, the mean values ​​were obtained from five mice each, and individual results are represented by hollow circles. One-way ANOVA and Turkey's test were used as post-hoc tests to determine the significance between individual experimental groups. Differences between experimental groups were considered significant, indicated by an * (P<0.05).

[0156]

[0157] Example 1. Confirmation of the inhibitory effects of sesamin on IL-2 production, CD69 expression, and proliferation in activated T cells.

[0158] To determine whether sesamin can modulate T cell function, we examined its effect on IL-2 mRNA expression. As shown in Figure 1A, sesamin significantly and dose-dependently reduced IL-2 mRNA expression, an indicator of activation, in human T cells stimulated with immobilized anti-CD3 and anti-CD28 antibodies (left), and also significantly reduced IL-2 mRNA expression in mouse CD4+ cells (right). Furthermore, as shown in Figure 1B, we confirmed that sesamin dose-dependently inhibited IL-2 production in human T cells and mouse CD4+ cells stimulated in conjunction with the T cell receptor (TCR) using ELISA. In particular, as confirmed in Fig. 1C, 20 to 40 μM sesamin significantly reduced the level of CD69 expression in activated T cells, and as shown in Fig. 1D, the overall intensity of CFSE fluorescence was reduced in proliferating T cells treated, and in particular, it was confirmed that CFSE fluorescence was blocked when 40 μM sesamin was treated.

[0159] In addition, as shown in Fig. 2A, the inhibitory effect of sesamin on IL-2 production in T cells activated by PMA / A23187 stimulation was confirmed, and the optimal concentration was 40 μM. In Fig. 2B, a time-dependent experiment also confirmed the inhibitory effect of 40 μM sesamin on IL-2 production in activated T cells.

[0160] Therefore, we confirmed that sesamin inhibits IL-2 production and CD69 expression in activated human and mouse T cells, and suppresses their function and proliferation.

[0161]

[0162] Example 2. Confirmation of physical interaction between MCL-1 active site of T cells and sesamin.

[0163] In order to confirm the basic mechanism of the T cell function inhibitory effect of sesamin, genes related to immune response were listed through the Genecards data bank. As a result, as shown in Figures 3A and 3B, 20,152 genes were classified as immune-related genes (a in Figure 3B), and 17,666 genes among them were confirmed to be involved in T cell function (b in Figure 3B). In addition, among the 17,666 genes, genes involved in the physiological activity of lignan compounds were confirmed, and 287 genes were confirmed as lignan-related genes (c in Figure 3B).

[0164] Among the 287 genes above, in order to identify proteins predicted as binding partners of sesamin, target proteins were predicted using the Swisstargetprediction database based on protein structures. As a result, as shown in Figure 5A, 20 proteins were identified as candidates for binding partners of sesamin in T cells, and three candidates were identified when evaluated with a cutoff value of 0.1 or higher in probability. As a result of confirming the affinity energy, Ki value, and ligand efficiency through molecular docking analysis, as shown in Figures 5B and 5C, among the three candidates, MCL-1 was confirmed to have the highest affinity energy (10.2 kcal / mol), the lowest Ki value (33.37 nM), and the highest ligand efficiency (0.39), and was ultimately selected as the target protein of sesamin.

[0165] In addition, as shown in Figures 4A to 4D, the molecular docking results confirmed that sesamin exhibited an affinity energy of 10.2 kcal / mol, which is almost similar to that of Umi-77, an MCL-1 inhibitor. Therefore, it was predicted that sesamin would be precisely located at the active site of MCL-1, where it binds to inhibit MCL-1 activity.

[0166] Furthermore, to confirm whether sesamin physically binds to MCL-1 in T cells, a pull-down assay using the CNBr-Sepharose 4B bead system was performed. As shown in Fig. 3C, MCL-1 in T cells was confirmed to be precipitated with Sepharose 4B beads (SS-beads) conjugated to sesamin in a dose-dependent manner. In the presence of soluble sesamin as a competitive inhibitor, the level of precipitated MCL-1 was confirmed to be significantly reduced when Sepharose 4B beads were mixed with the lysate. In contrast to MCL-1 precipitation, Bak and Bax, partner proteins that form heterodimers with MCL-1, were not detected in the pull-down assay using SS-beads.

[0167] Through this, it was confirmed that sesamin physically interacts with the active site of MCL-1 in T cells, and MLC-1 was confirmed to be the target molecule of sesamin.

[0168]

[0169] Example 3. Confirmation of the effect of sesamin on suppressing MCL-1 activity in activated T cells and regulating T cell activation through this effect.

[0170] To assess whether the interaction between sesamin and MCL-1 affects MCL-1 activity in T cells, we confirmed the expression of phosphorylated MCL-1 at the Ser64 site, which has already been demonstrated to be an indicator of MCL-1 activity.

[0171] As shown in Figure 6A, Western blotting results confirmed that MCL-1 expression was increased by TCR-mediated stimulation of human T cells, and that MCL-1 was completely phosphorylated within 6 hours. In contrast, treatment with 40 μM sesamin significantly inhibited MCL-1 phosphorylation at Ser64, similar to treatment with Umi-77. In addition, as a result of confirming the inhibitory effect of sesamin on MCL-1 phosphorylation in mouse CD4+ T cells, as shown in Figure 6B, it was confirmed that sesamin significantly down-regulated the phosphorylation level of MCL-1 without affecting the level of MCL-1 expression.

[0172] In addition, to verify the regulatory effect of sesamin on MCL-1 activity in T cells, a co-treatment analysis was performed using Umi-77. As shown in Fig. 6C, pretreatment with Umi-77 and 40 μM sesamin showed a greater inhibitory effect on MCL-1 activity than treatment with Umi-77 alone.

[0173] Furthermore, we confirmed by ELISA whether the decrease in MCL-1 activity by sesamin and Umi-77 treatment also caused downregulation of T cell function. As shown in Fig. 6D, simultaneous treatment with 40 μM sesamin and Umi-77 significantly reduced IL-2 production by activated T cells. Furthermore, as shown in Fig. 6E, pretreatment with Umi-77 and 40 μM sesamin showed a more superior proliferation inhibitory effect on activated T cells than pretreatment with Umi-77 alone.

[0174] Through this, we confirmed that sesamin regulates human and mouse T cell activation by inhibiting MCL-1 activity, and that its effect is enhanced when co-treated with Umi-77.

[0175]

[0176] Example 4. Confirmation of the effect of sesamin on blocking heterodimerization of Bak and MCL-1 in activated T cells.

[0177] Since MCL-1 plays a pivotal role in regulating the apoptotic pathway by forming heterodimers with Bak in various cell types, we examined the expression of Bcl-2 family proteins, including MCL-1, Bak, Bax, and Bcl, in resting and activated T cells. As shown in Figure 7A, MCL-1 expression was upregulated, whereas the expression of Bak, Bax, and Bcl-2 was unaffected by TCR stimulation or sesamin pretreatment.

[0178] Based on this, co-immunoprecipitation (co-IP) experiments were performed to determine whether T cell stimulation and pretreatment with 40 μM sesamin affect the interaction between MCL-1 and Bak. As shown in Figure 7B, the interaction between MCL-1 and Bak was significantly downregulated by stimulation, regardless of the presence or absence of sesamin. In particular, treatment with sesamin significantly reduced heterodimerization between MCL-1 and Bak in activated T cells.

[0179] Through this, we confirmed that inhibition of MCL-1 activity by sesamin in activated T cells blocks heterodimerization of Bak and MCL-1, thereby affecting the apoptosis pathway of activated T cells.

[0180]

[0181] Example 5. Confirmation of the specific and selective apoptosis-inducing effect of sesamin on activated T cells.

[0182] Pretreatment with 40 μM sesamin confirmed that the heterodimerization between MCL-1 and the pro-apoptotic protein Bak was reduced in activated T cells. In other words, it was confirmed that sesamin binds to the active site of MCL-1, inhibits its activity, reduces dimerization with pro-apoptotic proteins, and accelerates the apoptotic pathway of T cells. Based on this, to further understand the physiological function of sesamin in activated T cells, the effect of sesamin on activation-induced cell death (AICD) by TCR ligation was investigated. As shown in Fig. 8A, the AnnexinV / 7AAD analysis results showed that TCR-mediated stimulation induced apoptosis after 48 h, but pretreatment with 20 μM and 40 μM sesamin enhanced the apoptotic population in both the early (AnnexinV+7AAD-) and late (AnnexinV+7AAD+) stages. Notably, the apoptotic population in resting T cells was not affected by 40 μM sesamin. In addition, Western blotting results, as shown in Figure 8B, confirmed that pretreatment with 20 μM and 40 μM sesamin significantly increased the cleavage of caspase-3 and caspase-8 in T cells 48 hours after activation. In addition, WST-8 analysis results, as shown in Figure 8C, confirmed that pretreatment with sesamin significantly reduced the downregulated viability of activated T cells, and the effect was dose-dependent. In addition, to confirm the viability of T cells during ACID, trypan blue exclusion method was performed, and as shown in Figure 8D, pretreatment with 40 μM sesamin promoted the apoptosis of activated T cells for 72 hours. The increased population of blue-stained T cells was also confirmed in activated T cells pretreated with sesamin.

[0183] In this regard, as shown in Figure 9, the results of a time-dependent experiment confirmed that the apoptotic population was upregulated by sesamin only in activated T cells. Furthermore, as shown in Figures 10A and 10B, when activated human T cells and mouse CD4+ T cells were pretreated with 40 μM sesamin, the cleavage of caspase-3 and caspase-8 was enhanced in a time-dependent manner.

[0184] Therefore, we confirmed that sesamin induces apoptosis only in activated human and mouse T cells, but not in resting T cells.

[0185]

[0186] Example 6. Confirmation of the therapeutic effect of sesamin in an animal model of atopic dermatitis.

[0187] 6-1. Clinical evaluation

[0188] To evaluate the actual therapeutic effect of sesamin on atopic dermatitis, sesamin was orally administered to animal models induced with atopic dermatitis through treatment with HDM extract and DNCB, and the ear tissue condition was monitored for 28 days. As a result, as shown in Figure 11A, the ear tissue of the AD experimental group showed typical AD symptoms such as redness, swelling, crusting, and lichenification. On the other hand, the experimental group administered orally sesamin showed significant alleviation of the AD symptoms, and showed a significant therapeutic effect in a dose-dependent manner, especially compared to the experimental group administered orally tofacitinib, which was the positive control group. In addition, when the ear thickness of each experimental group was evaluated using clinical scores, it was confirmed that the ear tissue thickness of the experimental group administered orally sesamin was significantly reduced, as shown in Figure 11B. In particular, the experimental group administered orally 25 mg / kg of sesamin showed approximately 60% of the ear thickness of the AD experimental group on day 28. Furthermore, as shown in Figure 11C, when examining body weight changes on day 28, it was confirmed that AD-induced weight loss was significantly alleviated by oral sesamin administration. Furthermore, as shown in Figure 10D, when evaluating the number of ear scratches per minute on day 28, oral sesamin administration also significantly reduced the number of scratches.

[0189] Therefore, it was clearly confirmed that sesamin has an actual atopic dermatitis treatment effect.

[0190]

[0191] 6-2. Histopathological analysis evaluation

[0192] Furthermore, ear tissue was stained with H&E to analyze changes in skin structure, cellularity, and inflammation. As shown in Figures 12A and 12B, oral sesamin administration alleviated the inflammatory response and significantly reduced the thickness of the epidermis and dermis. Furthermore, as shown in Figure 12C, oral sesamin administration also attenuated inflammation and dermal papillae, typical structural changes observed in AD.

[0193] In addition, since infiltrated granule-positive mast cells play an important role in the development of AD, the number of infiltrated granule-positive mast cells was significantly reduced by oral administration of sesamin when ear tissues were stained with toluidine blue, as shown in Figures 12D and 12E. In addition, when we activated degranulation of mast cells during an allergic reaction and checked the serum level of IgE, a characteristic of AD, we confirmed that the serum IgE level was reduced by oral administration of sesamin, as shown in Figure 12F.

[0194] This confirmed that oral administration of sesamin attenuated the pathological manifestations of AD in vivo, including structural changes and mast cell infiltration.

[0195]

[0196] 6-3. Analysis of gene expression related to atopic dermatitis

[0197] Using qPCR analysis, we examined the effect of oral sesamin administration on the expression of pathogenic genes associated with AD symptoms. As a result, as shown in Figure 13A, the oral sesamin administration experimental group showed lower expression of Th2 cytokine mRNA, including il4, il5, il13, and il31, compared to the AD experimental group. Furthermore, as shown in Figures 13B and 13C, oral sesamin administration also reduced the mRNA expression of Th1 cytokines (ifng and tnfa) and Th17 cytokine (il17). On the other hand, as shown in Figures 14A and 14B, the expression of il6 and keratinocyte chemokine (tslp) was not affected.

[0198] Through this, it was confirmed that oral administration of sesamin significantly reduced the expression level of pathogenic genes in ear tissue in an AD-induced animal model.

[0199]

[0200] 6-4. Confirmation of improved systemic immune response

[0201] To determine whether oral administration of sesamin affects systemic T cell immunity during AD development, we compared the weight and length of the draining lymph nodes (dLNs), which are cervical lymph nodes, and the inguinal lymph nodes (non-dLNs). As a result, as shown in Figure 15A, in the AD experimental group, the dLNs were confirmed to be swollen and enlarged. On the other hand, as shown in Figure 15B, the weight and length of the dLNs were confirmed to be reduced when sesamin was administered orally. However, as shown in Figure 15C, no significant morphological changes were observed in non-dLNs despite oral sesamin administration, and as shown in Figure 15D, no changes were observed in the weight or length of non-dLNs. To determine whether oral administration of sesamin can improve the systemic immune response in AD, we examined the mRNA expression of AD-related genes in dLN tissues. As a result, as shown in Figure 15E, when sesamin was administered orally, the mRNA levels of Th2 cytokines including il4, il5, and il13 and the Th2 master transcription factor gata3 were significantly reduced.

[0202] Through this, it was confirmed that oral administration of sesamin systematically improved T cell-mediated immune responses in an AD-induced animal model.

[0203]

[0204] 6-5. Confirmation of the effect of regulating MCL-1 activity and promoting apoptosis mechanisms in vivo.

[0205] In addition, we confirmed the levels of phosphorylated MCL-1, cleaved caspase-3, and cleaved caspase-8 in dLN tissue. As a result, as shown in Figure 16A, the increased expression of phosphorylated MCL-1 induced by AD was reduced by oral sesamin administration, but the increased expression of cleaved caspase-3 and caspase-8 induced by AD was increased. Accordingly, protein expression in ear tissue was evaluated and whether oral sesamin administration affected cells in skin tissue. As a result, as shown in Figure 16B, the increased expression of phosphorylated MCL-1 induced by AD was downregulated by oral sesamin administration, but the expression of cleaved caspase-3 and cleaved caspase-8 induced by AD was upregulated.

[0206] Through this, we confirmed that oral administration of sesamin induces apoptosis by regulating MCL-1 activity in systemic immune tissues as well as local tissues in an AD-induced animal model.

[0207]

[0208] As described above, specific embodiments of the present invention have been described in detail. However, those skilled in the art who understand the spirit of the present invention will be able to easily suggest other inventions that are backwards or other embodiments included within the scope of the spirit of the present invention by adding, changing, or deleting other components within the scope of the same spirit. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention is indicated by the scope of the claims described below rather than the detailed description described above, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. A pharmaceutical composition for preventing or treating atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

2. In paragraph 1, A pharmaceutical composition wherein the above sesamin is represented by the following chemical formula 1. [Chemical Formula 1] 3. In paragraph 1, A pharmaceutical composition characterized in that the above sesamin inhibits at least one selected from the group consisting of activation, function and proliferation of T cells.

4. In paragraph 1, A pharmaceutical composition characterized in that the above sesamin promotes apoptosis of activated T cells.

5. In paragraph 1, A pharmaceutical composition characterized in that the above sesamin inhibits MCL-1 of activated T cells.

6. In paragraph 1, A pharmaceutical composition characterized in that the above sesamin increases a T cell-mediated immune response.

7. In paragraph 1, A pharmaceutical composition, wherein the pharmaceutical composition further comprises a Bcl-2 inhibitor.

8. In paragraph 7, A pharmaceutical composition characterized in that the Bcl-2 inhibitor is at least one selected from the group consisting of UMI-77, obatoclax, navitoclax, venetoclax, sabutoclax, A-1155463, A-1331852, ABT-737, TW-37, A-1210477, AT101, HA14-1, BAM7, S44563 and pharmaceutically acceptable salts thereof.

9. In paragraph 1, A pharmaceutical composition, characterized in that the pharmaceutical composition is administered orally.

10. A non-prescription drug for preventing or improving atopic dermatitis containing sesamin or a pharmaceutically acceptable salt thereof.

11. A cosmetic composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

12. A food composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

13. A health functional food composition for preventing or improving atopic dermatitis, comprising sesamin or a pharmaceutically acceptable salt thereof.

14. A method for preventing or treating atopic dermatitis, comprising the step of administering sesamin or a pharmaceutically acceptable salt thereof to a subject.

15. In paragraph 14, A method for preventing or treating atopic dermatitis, wherein the above object is additionally administered with a Bcl-2 inhibitor.

16. In paragraph 15, A method for preventing or treating atopic dermatitis, characterized in that the Bcl-2 inhibitor is at least one selected from the group consisting of UMI-77, obatoclax, navitoclax, venetoclax, sabutoclax, A-1155463, A-1331852, ABT-737, TW-37, A-1210477, AT101, HA14-1, BAM7, S44563 and pharmaceutically acceptable salts thereof.

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