Composition containing eclipta prostrata extract-derived compound as active ingredient for preventing, alleviating or treating pemphigus

Abstract

The present invention relates to a composition containing an Eclipta prostrata extract-derived compound as an active component for preventing, alleviating or treating pemphigus. The compound of Chemical Formula 1, which is the compound derived from an Eclipta prostrata extract of the present invention, has almost no cytotoxicity and exhibits antioxidant activity (reduction of reactive oxygen species (ROS) generation), and an excellent inhibitory effect on keratinocyte dissociation induced by AK23 (Anti-Desmoglein 3), thereby being able to be usefully used for preventing, alleviating or treating pemphigus.

Description

Composition for preventing, improving or treating pemphigus containing a compound derived from the extract of Korean lily as an active ingredient

[0001] The present invention relates to a composition for preventing, improving or treating pemphigus, containing a compound derived from an extract of Korean lily as an active ingredient.

[0002] This invention was carried out with the support of the project 'Development of herbal medicine application technology to solve industrial demand (Research on advanced Korean medicine treatment technology for rare and intractable diseases using transgenic zebrafish models)' (Project No.: 1711195904; KSN20233302), a basic project of the Korea Institute of Oriental Medicine under (or affiliated with) the Ministry of Science and ICT.

[0003] Pemphigus is a chronic, blistering disease that causes blisters on the skin and mucous membranes. Histologically, it is characterized by acantholysis, a phenomenon in which epithelial cell-cell junctions are disrupted. Most patients harbor autoantibodies in their serum against antigens present on the surface of epithelial cells. Because autoantibodies are directly involved in the pathogenesis of pemphigus, it is considered a representative autoimmune disease caused by autoantibodies.

[0004] Depending on clinical and histological findings, pemphigus can be divided into pemphigus vulgaris, proliferative pemphigus, pemphigus foliaceus, and pemphigus erythematosus. Among these, pemphigus vulgaris is the most common type of pemphigus, occurring equally in men and women and frequently in middle-aged people. In the case of pemphigus vulgaris, erosions occur in the oral mucosa in almost all patients, and oral lesions are the first symptoms in most patients. It frequently occurs in the buccal mucosa, palate, gums, and tongue of the oral cavity, and rather than vesicular lesions, they usually rupture and reveal themselves as cloudy spots caused by erosions or shed epithelial tissue. When the affected area is extensive, even minor stimulation can cause severe pain, causing great inconvenience in eating, and if it persists for a long time, it may induce findings such as hypoproteinemia, weight loss, or anemia. In some patients, only a few oral lesions develop locally, with the disease progressing for years with periods of worsening and improvement. In other cases, oral lesions remain as refractory residual lesions even after extensive skin lesions have resolved with treatment. Blisters typically begin to appear on the skin about four months after the oral lesions appear, frequently appearing on the face, trunk, groin, and axilla. Most pemphigus vulgaris begins with a few lesions in a localized area such as the oral cavity, but after several months, widespread lesions can develop throughout the body, and if left untreated, death can occur. Therefore, early treatment with sufficient oral steroids when the lesions are localized is essential to prevent further progression.

[0005] Treatment for pemphigus typically involves systemic steroid administration for a period of time, immunosuppressants, or plasmapheresis. Adjunctive therapies such as antibiotics, vitamins, and antihistamines are also recommended. While systemic steroids are the primary treatment for pemphigus, long-term use can lead to adverse effects. Immunosuppressants, while used in combination with steroids, can also cause side effects such as thrombocytopenia, infection, hemorrhagic cystitis, infertility, digestive problems, hepatitis, and hair loss. Therefore, there is a pressing need to develop natural products that can overcome the shortcomings of existing treatments and effectively improve pemphigus without side effects.

[0006] Meanwhile, as a technology related to pemphigus, Korean Patent Publication No. 2024-0030407 discloses a composition for improving or treating pemphigus containing an MKK3 kinase inhibitory compound, and Korean Patent Registration No. 1769858 discloses a technology for treating pemphigus containing an anti-Fas ligand antibody. However, a composition for preventing, improving or treating pemphigus containing the compound derived from the extract of Korean angelica tree of the present invention as an active ingredient has not yet been disclosed.

[0007] The present invention was derived from the above-mentioned needs, and provides a composition for preventing, improving or treating pemphigus containing a compound derived from an extract of Korean ginseng as an active ingredient, and the compound of chemical formula 1, which is a compound derived from an extract of Korean ginseng, not only has almost no cytotoxicity, but also has antioxidant activity (reducing the amount of reactive oxygen species (ROS) produced), and has an excellent effect of inhibiting the dissociation of keratinocytes induced by AK23 (Anti-Desmoglein 3), thereby completing the present invention.

[0008] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating pemphigus, which contains a compound of chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0009] In addition, the present invention provides a health functional food composition for preventing or improving pemphigus, which contains a compound of chemical formula 1 or a food-wise acceptable salt thereof as an active ingredient.

[0010] In addition, the present invention provides an over-the-counter drug for preventing or improving pemphigus, which contains a compound of chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0011] In addition, the present invention provides a veterinary composition for preventing or treating pemphigus, which contains a compound of chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0012] In addition, the present invention provides a feed additive for preventing or improving pemphigus, which contains a compound of chemical formula 1 or a food-wise acceptable salt thereof as an active ingredient.

[0013] In addition, the present invention provides a cosmetic composition for preventing or improving pemphigus, which contains a compound of chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0014] The present invention relates to a composition for preventing, improving or treating pemphigus, containing a compound derived from an extract of Korean lily as an active ingredient. The compound of chemical formula 1, which is a compound derived from an extract of Korean lily of the valley of the present invention, not only has almost no cytotoxicity, but also has antioxidant activity (reducing the amount of reactive oxygen species (ROS) produced), and is excellent in inhibiting the dissociation of keratinocytes induced by AK23 (Anti-Desmoglein 3).

[0015] Figure 1 shows the results of treating HaCaT cells with compounds derived from the extract of the present invention (compounds of chemical formula 1) [pratensein (A) and pratensein-7-O-β-D-glucopyranoside (pratensein-G) (B)] at various concentrations and checking cell viability. *, **, ***, **** indicate that cell viability was statistically significantly increased in the group treated with compounds derived from the extract of the Korean ginseng compared to the untreated group, with p<0.05, p<0.01, p<0.001, and p<0.0001, respectively.

[0016] Figure 2 shows the results of confirming the effect of inhibiting keratinocyte dissociation according to treatment with the compound (compound of chemical formula 1) derived from the extract of the present invention [pratensein (A) and pratensein-7-O-β-D-glucopyranoside (pratensein-G) (B)] after inducing keratinocyte dissociation by treating with AK23. ## indicates that the dissociation of keratinocytes in the group in which pemphigus was induced by treating with AK23 was statistically significantly increased compared to the keratinocytes that were not treated with anything, and p<0.01, and **, *** indicate that the dissociation of keratinocytes in the group in which the compound of chemical formula 1, which is an active ingredient of the present invention, was treated was statistically significantly decreased compared to the pemphigus-induced group, and ** is p<0.01, and *** is p<0.001. ns means that the difference between the comparison groups is not statistically significant.

[0017] Figure 3 shows the results confirming that the increased ROS production induced by LPS in macrophages was reduced by treatment with the compounds (compounds of chemical formula 1) [Pratensein and Pratensein-G] derived from the extract of the Korean angelica tree of the present invention. ## indicates that the ROS production in the LPS-treated group was statistically significantly increased compared to untreated macrophages, and p<0.01, and * indicates that the ROS production in the group treated with the compound of chemical formula 1 (pratensein), which is the active ingredient of the present invention, was statistically significantly reduced compared to the LPS-treated group, and p<0.05. ns indicates that the difference between the comparison groups is not statistically significant.

[0018] Figure 4 is a photograph (A) confirming that the increased ROS production induced by LPS in zebrafish larvae was reduced by treatment with compounds (compounds of chemical formula 1) [pratensein and pratensein-G] derived from the extract of the Korean angelica tree of the present invention, and a graph (B) quantifying the same. ### indicates that the ROS production in the LPS-treated group was statistically significantly increased compared to zebrafish larvae that were not treated with anything, and p<0.001, and *** indicates that the ROS production in the group treated with the compounds of chemical formula 1 (pratensein and pratensein-G), which are the active ingredients of the present invention, was statistically significantly reduced compared to the LPS-treated group, and p<0.001.

[0019] The present invention relates to a pharmaceutical composition for preventing or treating pemphigus, which contains a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0020]

[0021] In the above chemical formula 1, R1 is H or glucose. In the above chemical formula 1, when R1 is H, it is named Pratensein, and when R1 is glucose, it is named Pratensein-G.

[0022] The compound of the above chemical formula 1 may be derived from an extract of Korean lily, but is not limited thereto.

[0023] As used herein, the term "acceptable salt" refers to any salt of a compound of the present invention that maintains the biological properties of the compound of the present invention while not exhibiting toxicity or being unsuitable for food or pharmaceutical use. Such salts may be derived from, and include, various organic and inorganic counterions known in the art. These salts include those obtained from organic or inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfamic acid, acetic acid, trifluoroacetic acid, trichloroacetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, glutaric acid, pyruvic acid, lactic acid, malonic acid, succinic acid, sorbic acid, ascorbic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, citric acid, cinnamic acid, mandelic acid, phthalic acid, lauric acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-Toluenesulfonic acid, camphoric acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-Oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, cyclohexylsulfamic acid, quinic acid or muconic acid; Or it may include salts formed when an acidic proton present in the parent compound is replaced by a metal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or an aluminum ion), an alkali metal or alkaline earth metal hydroxide (e.g., sodium, potassium, calcium, magnesium, aluminum, lithium, zinc, and barium hydroxide), ammonia, or coordinated with an organic base, such as an aliphatic, alicyclic, or aromatic organic amine, for example, ammonia, methylamine, dimethylamine, diethylamine, picoline, ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylene-diamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, N-methylglucamine piperazine, tris(hydroxymethyl)-aminomethane, or tetramethylammonium hydroxide.

[0024] In addition to the above-mentioned active ingredients, the pharmaceutical composition may further include a pharmaceutically acceptable carrier, excipient, or diluent, and the pharmaceutical composition may be formulated for systemic administration or manufactured as a formulation for topical administration. The pharmaceutical composition may be formulated as an injection administered by injection; or as an oral administration agent in the form of a tablet, capsule, liquid, suspension, or powder; and the pharmaceutical composition may be manufactured as a gel, hydrogel, ointment, cream, foam, spray, aerosol, lotion, liquid, or skin patch, but is not limited thereto. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include capsules, powders, granules, tablets, pills, 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.

[0025] In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, and aerosols. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizers, and suppositories. Non-aqueous solvents and suspending agents can be used, such as propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can be witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin. When administering parenterally, it is advisable to choose topical application on the skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine, epidural, or intracerebrovascular injection.

[0026] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The level of the effective amount may be determined based on the type and severity of the patient's disease, the activity and sensitivity of the drug, the time of administration, the route of administration, and the excretion rate, the duration of treatment, concomitant drugs, and other factors well known in the medical field.

[0027] In addition, the present invention relates to a health functional food composition for preventing or improving pemphigus, which contains a compound of the following chemical formula 1 or a food-wise acceptable salt thereof as an active ingredient.

[0028] [Chemical Formula 1]

[0029]

[0030] In the above chemical formula 1, R1 is H or glucose.

[0031] The above composition is preferably prepared in any one of the following forms: powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto.

[0032] When the health functional food composition of the present invention is used as a food additive, the effective ingredient may be added as is or used together with other foods or food ingredients, and may be used appropriately according to a conventional method. The amount of the effective ingredient mixed may be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, when consumed for the purpose of health and hygiene or for the purpose of health control for a long period of time, the amount may be below the above range, and since there is no problem in terms of safety, the effective ingredient may also be used in an amount exceeding the above range.

[0033] There are no specific restrictions on the types of the above foods. Examples of foods to which the above active ingredients can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and all health functional foods in the conventional sense are included.

[0034] When the composition of the present invention is used as a health beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in conventional beverages. The natural carbohydrates mentioned above are 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. As a sweetener, a natural sweetener such as thaumatin or stevia extract, or a synthetic sweetener such as saccharin or aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 g of the composition of the present invention.

[0035] The composition of the present invention may contain 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 contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in mixtures. The proportion of these additives is not particularly important, but the composition of the present invention is typically selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

[0036] In addition, the present invention relates to an over-the-counter drug for preventing or improving pemphigus, which contains a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0037] [Chemical Formula 1]

[0038]

[0039] In the above chemical formula 1, R1 is H or glucose.

[0040] In the present invention, the term 'quasi-drug' means a fiber, rubber product or the like used for the purpose of treating, alleviating, managing or preventing a disease in humans or animals; a thing that has a weak effect on the human body or does not directly affect the human body, and is not a device or machine and the like; an article corresponding to one of the preparations used for sterilization, insecticide and the like for the prevention of infectious diseases; an article used for the purpose of diagnosing, treating, alleviating, managing or preventing a disease in humans or animals, other than an device, machine or device; and an article used for the purpose of exerting a pharmacological effect on the structure and function of humans or animals, excluding an article that is not an device, machine or device. In addition, the above-mentioned quasi-drug includes external skin preparations and personal hygiene products. When the compound of Chemical Formula 1 of the present invention is used as an quasi-drug additive, the compound of Chemical Formula 1 may be added as is or used together with another quasi-drug or quasi-drug ingredient, and may be used appropriately according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the intended use. The above skin external preparation is not particularly limited thereto, but preferably can be manufactured and used in the form of ointment, lotion, spray, patch, cream, powder, suspension, gel or gel. The above personal hygiene products are not particularly limited thereto, but preferably can be soap, wet tissue, toilet paper, skin cream, face cream, air freshener gel or cleansing gel. In addition, other examples of the non-medicinal product of the present invention include disinfectant, shower foam, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment or filter filler.

[0041] In addition, the present invention relates to a veterinary composition for the prevention or treatment of pemphigus, which contains a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0042] [Chemical Formula 1]

[0043]

[0044] In the above chemical formula 1, R1 is H or glucose.

[0045] The veterinary composition of the present invention may further comprise suitable excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition of the present invention 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, ethanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil. The veterinary composition according to the present invention may further include fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc., and the veterinary composition according to the present invention may be formulated using a method well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to an animal, and the formulation may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile topical preparations, etc. The effective amount of the veterinary composition according to the present invention may be appropriately selected depending on the individual animal. It may be determined according to factors including the severity of the disease or condition, the sensitivity to the active ingredient of the present invention depending on the age, weight, health condition or sex of the individual, the route of administration, the period of administration, other compositions combined with or used simultaneously with the composition, and other factors well known in the physiological or veterinary fields.

[0046] In addition, the present invention relates to a feed additive for preventing or improving pemphigus, which contains a compound of the following chemical formula 1 or a food-wise acceptable salt thereof as an active ingredient.

[0047] [Chemical Formula 1]

[0048]

[0049] In the above chemical formula 1, R1 is H or glucose.

[0050] The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. The term "feed" in the present invention may mean any natural or artificial diet, meal, etc., or ingredients of the meal, which are intended for or suitable for animals to eat, ingest, and digest. The type of the feed is not particularly limited, and feed commonly used in the relevant technical field may be used. Non-limiting examples of the feed include plant feeds such as grains, roots, fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal, or grain by-products; and animal feeds such as proteins, inorganic substances, oils and fats, mineral substances, oils and fats, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0051] In addition, the present invention relates to a cosmetic composition for preventing or improving pemphigus, which contains a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0052] [Chemical Formula 1]

[0053]

[0054] In the above chemical formula 1, R1 is H or glucose.

[0055] The cosmetic composition of the present invention includes, in addition to the above-mentioned effective ingredients, components commonly used in cosmetic compositions, such as fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, and conventional auxiliary agents and carriers such as lipid vesicles.

[0056] The cosmetic composition of the present invention can be prepared in any formulation commonly manufactured in the art, and can be formulated as, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray. More specifically, the cosmetic composition of the present invention is a skin, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nourishing lotion, massage cream, nourishing cream, eye cream, moisturizing cream, hand cream, essence, nourishing essence, pack, cleansing foam, cleansing water, cleansing lotion, cleansing cream, body lotion, body cleanser, soap, powder, hair tonic, hair conditioner, hair essence, hair lotion, hair nourishing lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nourishing cream, hair moisture cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nourishing pack, hair soap, hair cleansing foam, hair drying agent, hair preservative, hair dye, hair waving agent, hair bleaching agent, hair gel, hair glaze, hair dressing, hair lacquer, hair moisturizer, hair mousse. Or it can be manufactured as a cosmetic formulation of hair spray, but is not limited thereto. When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide can be used as a carrier component. When the formulation of the cosmetic composition of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component, and particularly in the case of a spray, it can additionally include a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether.When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, a solubilizer or an emulsifier 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. When the formulation of the cosmetic composition of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, and the like can be used as a carrier component.

[0057]

[0058] Hereinafter, the present invention will be described in more detail using examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0059]

[0060] Manufacturing Example 1. Separation and identification of the compound of chemical formula 1 from the extract of Korean lily.

[0061] The compound of chemical formula 1 of the present invention was used as provided by Kyunghee University, and the provided compound of chemical formula 1 was separated from a 70% ethanol extract of Eclipta prostrata using HPLC, and then, 1 H-NMR, 13 The compound of chemical formula 1 was identified through C-NMR and mass spectrometry and confirmed to be Pratensein and Pratensein-7-O-β-D-glucopyranoside.

[0062]

[0063] Example 1. Confirmation of cytotoxicity of compound of chemical formula 1

[0064] Human skin cells, HaCaT cells (AddexBio, CA, USA), were cultured in DMEM / high glucose conditions with 10% fetal bovine serum and 1% antibiotic-antimycotic solution at 37°C and 5% CO2. To determine cytotoxicity, 1 × 10 4 HaCaT cells were seeded and stabilized for one day. After confirming that the cells in each well had formed a monolayer, the cells were treated with the compound of chemical formula 1 and cultured for one day. Afterwards, the cell viability was confirmed by treating with CCK-8 reagent (Dojindo, USA).

[0065] As a result, as disclosed in Fig. 1, it was confirmed that the cell viability following treatment with the compound of chemical formula 1 was similar to or rather increased compared to the untreated group.

[0066]

[0067] Example 2. Confirmation of the effect of the compound of chemical formula 1 on inhibiting keratinocyte dissociation.

[0068] HaCaT cells were seeded at 1×10 per well in a well plate. 4 After inoculating with 10 cells and stabilizing for a day, it was confirmed that the cells formed a monolayer well in each well. Then, the compound of chemical formula 1 was treated at a concentration that was not toxic to the cells, and the antibody (MBL, Japan) was treated to each well at an amount of 5 μg / ml and cultured for a day. After that, each well was washed with PBS and incubated with 150 μl of Hanks' buffered saline (HBSS, Gibco) containing dispase II reagent (2.4 U / ml, Sigma) to separate the cell monolayer and the well plate at 37°C for 20 minutes. After that, 200 μl of Hanks' buffered saline was added, and the cell monolayer was exposed to mechanical stress by pipetting 15 times with a 1 ml pipette to shear it.

[0069] Next, the cells were stained with MTT (5 mg / ml; Sigma) for 20 min, and the number of generated fragments was captured by microscopy.

[0070] As a result, as disclosed in FIG. 2, the compound of chemical formula 1 of the present invention was excellent in inhibiting the dissociation of keratinocytes induced by AK23 treatment.

[0071]

[0072] Example 3. Evaluation of inhibition ability of reactive oxygen species (ROS) production using RAW 264.7 cells.

[0073] 2×10 per well 5 RAW 264.7 macrophages were seeded into 96-well microplates at a cell density of 10 and cultured for 4 hours in an incubator at 37°C and 5% CO₂ to allow cell attachment. Afterwards, test substances (placentaine, placentaine-G) were added to a final concentration of 5 μM each, and simultaneously treated with lipopolysaccharide (LPS) to a final concentration of 1 μg / mL and cultured for an additional 24 hours. After the incubation was completed, 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA, Sigma-Aldrich) was added to each well to a final concentration of 20 μM and the reaction was performed under dark conditions for 30 minutes. Afterwards, the fluorescence intensity of each well was measured using a multi-detection microplate reader (Bio-Tek, Winooski, VT, USA). Fluorescence measurements were performed based on an excitation wavelength of 485 nm and an emission wavelength of 528 nm. The measured fluorescence intensity indicates the amount of intracellular reactive oxygen species (ROS), and the ROS production inhibition effect of the test substance was quantitatively evaluated through this.

[0074] As a result, as disclosed in Fig. 3, the amount of intracellular reactive oxygen species (ROS) production increased by LPS treatment, and in contrast, it was confirmed that the amount of ROS production significantly decreased in the group treated with Pratensein and Pratensein-G, which are the effective ingredients of the present invention.

[0075]

[0076] Example 4. Evaluation of the ability to inhibit reactive oxygen species (ROS) production using zebrafish larvae.

[0077] The inhibitory effect on reactive oxygen species (ROS) production was evaluated in zebrafish larvae 3 days post-fertilization (3 dpf).

[0078] The experimental group was treated with lipopolysaccharide (LPS) at a final concentration of 0.5 mg / ml, and the negative control group was treated with phosphate-buffered saline (PBS) under the same conditions. The larvae of each experimental and control group were immersed in E3 culture medium containing the test substance at a final concentration of 10 μM. All larvae were cultured at 28.5°C for 24 hours. Afterwards, the larvae of each group were treated with 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) at a final concentration of 10 μM and reacted in the dark for 30 minutes. Fluorescence images were acquired using a fluorescence microscope with an excitation wavelength of 485 nm and an emission wavelength of 528 nm, and the acquired fluorescence intensity was quantitatively analyzed using Image J software. The fluorescence intensity of each experimental group was converted into a percentage (%) relative to the untreated group (negative control group) to compare the level of ROS production.

[0079] As a result, as disclosed in Fig. 4, the amount of reactive oxygen species (ROS) produced in zebrafish larvae increased due to LPS treatment, and in contrast, it was confirmed that the amount of ROS produced was significantly reduced in the group treated with Pratensein and Pratensein-G, which are the effective ingredients of the present invention.

Claims

1. A pharmaceutical composition for the prevention or treatment of pemphigus, containing a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical Formula 1] In the above chemical formula 1, R1 is H or glucose.

2. A pharmaceutical composition for the prevention or treatment of pemphigus, characterized in that the compound of the chemical formula 1 in the first paragraph is derived from an extract of Korean licorice.

3. A pharmaceutical composition for the prevention or treatment of pemphigus, characterized in that, in addition to the effective ingredient in paragraph 1, it further comprises a pharmaceutically acceptable carrier, excipient or diluent.

4. A pharmaceutical composition for the prevention or treatment of pemphigus, characterized in that the pharmaceutical composition in claim 1 is formulated for systemic administration.

5. A pharmaceutical composition for preventing or treating pemphigus, characterized in that the pharmaceutical composition in paragraph 1 is an injectable agent administered by injection; or an oral agent in the form of a tablet, capsule, liquid, suspension, or powder.

6. A pharmaceutical composition for the prevention or treatment of pemphigus, characterized in that the pharmaceutical composition in claim 1 is prepared in a form for topical administration.

7. A pharmaceutical composition for preventing or treating pemphigus, characterized in that the pharmaceutical composition in the first paragraph is in the form of a gel, hydrogel, ointment, cream, foam, spray, aerosol, lotion, liquid, or skin patch.

8. A health functional food composition for preventing or improving pemphigus, containing a compound of the following chemical formula 1 or a food-wise acceptable salt thereof as an active ingredient. [Chemical Formula 1] In the above chemical formula 1, R1 is H or glucose.

9. A health functional food composition for preventing or improving pemphigus, characterized in that the composition in the 8th paragraph is manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candy, syrup, and beverage.

10. A non-medicinal product for preventing or improving pemphigus, containing a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical Formula 1] In the above chemical formula 1, R1 is H or glucose.

11. A veterinary composition for the prevention or treatment of pemphigus, containing a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical Formula 1] In the above chemical formula 1, R1 is H or glucose.

12. A feed additive for preventing or improving pemphigus, containing a compound of the following chemical formula 1 or a food-wise acceptable salt thereof as an active ingredient. [Chemical Formula 1] In the above chemical formula 1, R1 is H or glucose.

13. A cosmetic composition for preventing or improving pemphigus, containing a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical Formula 1] In the above chemical formula 1, R1 is H or glucose.

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