Composition containing a fraction of Syzygium formosum extract as an active ingredient

Abstract

The present invention relates to a composition containing a fraction of a Syzygium formosum extract as an active ingredient. According to one embodiment, a composition containing a fraction of a Syzygium formosum extract as an active ingredient can be useful for preventing, improving, or treating inflammatory diseases by suppressing the expression of inflammatory cytokines, and has the effect of making cosmetics containing the composition highly stable.

Description

[Technical Field]

[0001] The present invention relates to a composition containing a fraction of an extract of Syzygium formosum as an active ingredient. [Background technology]

[0002] Inflammation is a localized defense response of the body in response to pathological conditions caused by physical trauma, harmful chemicals, infections by bacteria, fungi, or viruses, or irritants in the body's metabolic products. Inflammation is triggered by a variety of inflammatory mediators produced by damaged tissue and migrating cells. During an inflammatory response, plasma accumulates at the site of inflammation, diluting the toxic substances secreted by bacteria and increasing blood flow, resulting in symptoms such as erythema, pain, swelling, and fever. Under normal circumstances, the body neutralizes or removes pathogenic factors through the inflammatory response and regenerates damaged tissue, restoring normal structure and function. However, if this does not occur, it can progress to a disease state such as chronic inflammation.

[0003] Macrophages are the main cells responsible for innate immunity and are activated by many factors, such as cytokines and bacterial lipopolysaccharide (LPS). Activated macrophages produce not only inflammatory factors such as nitric oxide (NO) and prostaglandin E2 (PGE2), but also infectious cytokines such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1 (IL-1).

[0004] In addition, cyclooxygenase (COX) has hydroperoxidase (HOX) activity in addition to the COX function, and synthesizes the intermediates PGG2 and PGG2 from arachidonic acid, producing these compounds as PGE2, PGF2, PGD2, prostacyclin, and thromboxane A2 (TxA2). Of the two COX isoforms, COX-2 is rapidly induced during inflammatory responses and plays an important role in generating PGE2 and other cytokines (Weisz A., Biochem. J., 316:209-215, 1996; (Miller MJ et al., Mediators of inflammation, 4:387-396, 1995; Appleton L. et al., Adv. Pharmacol., 35:27-28, 1996). In addition to its well-known role as a mediator of inflammatory responses, PGE2 also suppresses the production of inflammatory cytokines such as TNF-α, IL-1β, IL-8, and IL-12 in macrophages.

[0005] Syzygium formosum is an evergreen tree native to Southeast Asia, including Bangladesh, India, Myanmar, Thailand, Laos, and Vietnam, and can grow to a height of 10 meters. In Vietnam and Laos, Syzygium formosum is cultivated and its fruit is used as food.

[0006] However, it has never been disclosed that a fraction of Syzygium formosum extract, as in the present invention, is effective as an anti-inflammatory agent. Summary of the Invention

[0007] One aspect is to provide a cosmetic composition comprising a fraction of an extract of Syzygium formosum as an active ingredient.

[0008] Another aspect of the present invention is to provide an external skin preparation composition for improving skin inflammation, which contains a fraction of a Syzygium formosum extract as an active ingredient.

[0009] Another aspect of the present invention is to provide a health functional food for improving skin inflammation, which contains a fraction of a Syzygium formosum extract as an active ingredient.

[0010] Another aspect is to provide a pharmaceutical composition for preventing or treating inflammatory skin diseases, which comprises a fraction of an extract of Syzygium formosum as an active ingredient.

[0011] Another aspect is to provide a method of preventing or treating skin inflammation comprising administering a fraction of an extract of Syzygium formosum to an individual in need thereof.

[0012] One aspect provides a cosmetic composition comprising a fraction of an extract of Syzygium formosum as an active ingredient.

[0013] The Syzygium formosum may be one or more selected from the group consisting of a whole plant, root, trunk, branch, leaf, seed, or fruit. According to one embodiment, the Syzygium formosum extract may be extracted using Syzygium formosum leaves. [Brief explanation of the drawings]

[0014] [Figure 1a] FIG. 1 is a graph showing the results of confirming cytotoxicity, where FIG. 1a is a graph showing the results of confirming cytotoxicity of Syzygium formosum extract, and FIG. 1b is a graph showing the results of confirming cytotoxicity of fractions of Syzygium formosum extract. [Figure 1b]FIG. 1 is a graph showing the results of confirming cytotoxicity, where FIG. 1a is a graph showing the results of confirming cytotoxicity of Syzygium formosum extract, and FIG. 1b is a graph showing the results of confirming cytotoxicity of fractions of Syzygium formosum extract. [Figure 2a] FIG. 2 is a graph showing the results of confirming the expression of COX-2 after UVB irradiation, where FIG. 2a is a graph showing the results of confirming the expression of COX-2 after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 2b is a graph showing the results of confirming the expression of COX-2 after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 2b] FIG. 2 is a graph showing the results of confirming the expression of COX-2 after UVB irradiation, where FIG. 2a is a graph showing the results of confirming the expression of COX-2 after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 2b is a graph showing the results of confirming the expression of COX-2 after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 3a] FIG. 3 is a graph showing the results of confirming the expression of IL-1β after UVB irradiation, where FIG. 3a is a graph showing the results of confirming the expression of IL-1β after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 3b is a graph showing the results of confirming the expression of IL-1β after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 3b] FIG. 3 is a graph showing the results of confirming the expression of IL-1β after UVB irradiation, where FIG. 3a is a graph showing the results of confirming the expression of IL-1β after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 3b is a graph showing the results of confirming the expression of IL-1β after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 4a] FIG. 4 is a graph showing the results of confirming the expression of IL-6 after UVB irradiation, where FIG. 4a is a graph showing the results of confirming the expression of IL-6 after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 4b is a graph showing the results of confirming the expression of IL-6 after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 4b]FIG. 4 is a graph showing the results of confirming the expression of IL-6 after UVB irradiation, where FIG. 4a is a graph showing the results of confirming the expression of IL-6 after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 4b is a graph showing the results of confirming the expression of IL-6 after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 5a] FIG. 5 is a graph showing the results of confirming the expression of IL-8 after UVB irradiation, where FIG. 5a is a graph showing the results of confirming the expression of IL-8 after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 5b is a graph showing the results of confirming the expression of IL-8 after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 5b] FIG. 5 is a graph showing the results of confirming the expression of IL-8 after UVB irradiation, where FIG. 5a is a graph showing the results of confirming the expression of IL-8 after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 5b is a graph showing the results of confirming the expression of IL-8 after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 6a] FIG. 6 is a graph showing the results of confirming the expression of TNF-α after UVB irradiation, where FIG. 6a is a graph showing the results of confirming the expression of TNF-α after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 6b is a graph showing the results of confirming the expression of TNF-α after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 6b] FIG. 6 is a graph showing the results of confirming the expression of TNF-α after UVB irradiation, where FIG. 6a is a graph showing the results of confirming the expression of TNF-α after UVB irradiation and treatment with Syzygium formosum extract, and FIG. 6b is a graph showing the results of confirming the expression of TNF-α after UVB irradiation and treatment with a fraction of Syzygium formosum extract. [Figure 7] FIG. 7 is a graph comparing IL-6 expression in cells treated with Centella asiatica extract, Syzygium formosum extract, or fractions of Syzygium formosum extract after UVB irradiation. [Figure 8]FIG. 8 is a graph comparing the expression of IL-1β in the cells treated with Centella asiatica extract, Syzygium formosum extract, or fractions of Syzygium formosum extract after UVB irradiation. [Figure 9] FIG. 9 is a graph comparing the expression of IL-8 in cells treated with Centella asiatica extract, Syzygium formosum extract, or fractions of Syzygium formosum extract after UVB irradiation. [Figure 10] FIG. 10 is a graph comparing the expression of COX-2 in the treated Centella asiatica extract, Syzygium formosum extract, or fractions of Syzygium formosum extract after UVB irradiation. [Figure 11] FIG. 11 is a graph showing the change in brightness of Syzygium formosum extract or a cream containing Syzygium formosum extract. [Figure 12] FIG. 12 is a graph showing the change in color of Syzygium formosum extract or a cream containing Syzygium formosum extract. [Figure 13] FIG. 13 is a graph showing the change in saturation of Syzygium formosum extract or a cream containing Syzygium formosum extract. [Figure 14] FIG. 14 is a graph showing the change in viscosity of Syzygium formosum extract or a cream containing Syzygium formosum extract. DETAILED DESCRIPTION OF THE INVENTION

[0015] As used herein, the term "active ingredient" or "effective amount" can refer to any amount of a composition used in the course of practicing the inventions provided herein that is sufficient to alleviate, inhibit the progression of, or prevent a disease, disorder, or condition, or one or more symptoms thereof.

[0016] As used herein, the term "fractionate" can refer to the product obtained by fractionation to separate a specific component or group of components from a mixture containing a variety of components.

[0017] The fractionation method for obtaining the fraction is not particularly limited and may be a method commonly used in the art. A non-limiting example of the fractionation method is a method for obtaining a fraction from an extract obtained by extracting Syzygium formosum and treating the extract with a predetermined solvent.

[0018] The type of fractionation solvent used to obtain the fraction is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the fractionation solvent include polar solvents such as water and alcohols such as butanol, and non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These solvents can be used alone or in combination. When alcohol is used as the fractionation solvent, C1 to C4 alcohols can be used.

[0019] The fraction of the Syzygium formosum extract may be a hexane fraction, an ethyl acetate fraction, a butanol fraction, or a water fraction of the Syzygium formosum extract, and may be obtained by repeating fractionation 1 to 5 times, for example, 2 to 5 times, 3 to 5 times, or 2 to 4 times. In one specific example of the fraction, the fraction may be a water fraction, and the fraction may be obtained by repeating fractionation 3 times.

[0020] The fractionation can include adding the fractionation solvent to the Syzygium formosum extract in an amount of 1 to 30 times (volume / weight), for example, 5 to 30 times (volume / weight), 5 to 20 times (volume / weight), 10 to 30 times (volume / weight), or 10 to 20 times (volume / weight).For example, the fractionation can include adding 100 ml to 3000 ml of the fractionation solvent to 100 g of the Syzygium formosum extract.

[0021] The fraction of Syzygium formosum extract contains triterpene compounds, such as asiatic acid, madecassic acid, colosolic acid, maslinic acid, betulinic acid, ursolic acid, and oleanolic acid, at higher concentrations than Syzygium formosum extract, and therefore has superior anti-inflammatory, anti-allergic, and skin regenerating effects compared to Syzygium formosum extract. Therefore, the cosmetic composition may be used for improving skin inflammation.

[0022] The term "skin inflammation" as used herein may refer to a disease induced by inflammatory cytokines, such as IL-6 (Interleukin-6) or TNF-α (Tumor necrosis factor-α), which are secreted from immune cells such as macrophages when the body's immune system is overactivated by harmful stimuli such as inflammatory factors or radiation. According to one embodiment, the composition reduces the activity of inflammatory cytokines, thereby providing anti-inflammatory, anti-allergic, or skin regenerating effects.

[0023] In one embodiment, the cosmetic composition may further comprise an emollient, a humectant, and a thickener.

[0024] The emollient may be selected from the group consisting of cetearyl alcohol, cetyl palmitate, beeswax, squalane, cetyl ethyl hexanoate, and caprylic / capric triglyceride; the humectant may be selected from the group consisting of 1,2-hexanediol, dipropylene glycol, and glycerin; and the thickener may be selected from the group consisting of xanthan gum and ammonium acryloyldimethyltaurate / VP copolymer.

[0025] Syzygium formosum extract contains large amounts of plant metabolites such as amino acids and sugars as well as triterpene compounds, and cosmetics containing the extract have problems such as color change, unstable viscosity, and emulsion instability. In contrast, the fraction of the Syzygium formosum extract contains high concentrations of triterpene compounds, and therefore cosmetics containing the extract have the effects of less color change, stable viscosity, and better stability.

[0026] The composition may comprise, relative to the total weight of the composition, 0.001% to 80% by weight of a fraction of a Syzygium formosum extract, e.g., 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, 0.01% to 20% by weight, 0.01% to 10% by weight, 0.01% to 5% by weight, 0.05% to 60% by weight, 0.05% to 40% by weight, 0.05% to 30% by weight, 0.05% to 20% by weight, 0.05% to 10% by weight, 0.05% to 5% by weight, 0.1% to 60% by weight, 0.1% to 40% by weight, 0.1% to 30% by weight, 0.1% to 20% by weight, 0.1% to 10% by weight, or 0.1% to 5% by weight.

[0027] The cosmetic composition may have a dosage form selected from the group consisting of softening lotion, nourishing lotion, nourishing cream, moisture cream, massage cream, essence, ampoule, gel, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, powder, gel, lotion, and ointment. The cosmetic composition may further contain one or more cosmetically acceptable carriers that are commonly incorporated into skin cosmetics, and may appropriately incorporate common ingredients such as, but not limited to, oil, water, surfactant, moisturizer, lower alcohol, thickener, chelating agent, colorant, preservative, and fragrance.

[0028] Another aspect provides a composition for external use on the skin for improving skin inflammation, which contains a fraction of a Syzygium formosum extract as an active ingredient.

[0029] The topical skin preparation may be a cream, gel, ointment, skin emulsion, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof.

[0030] The topical skin preparation may contain, as needed, ingredients used in topical skin preparations such as ordinary cosmetics and pharmaceuticals, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, and various skin nourishing agents.

[0031] The topical skin preparation may also contain, as appropriate, chelating agents such as edetate disodium, edetate trisodium, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; caffeine, tannin, verapamil, licorice extract, glabridin, hot water extract of quince fruit, various herbal medicines, drugs such as tocopherol acetate, glycyrrhizic acid, tranexamic acid and derivatives thereof or salts thereof; vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid, sugars such as glucose, fructose, and trehalose; and the like.

[0032] In still another embodiment, there is provided a health functional food for improving skin inflammation, which contains a fraction of a Syzygium formosum extract as an active ingredient.

[0033] The functional health food refers to any food that is produced using nutrients that are easily lacking in daily diets or raw materials or ingredients that have functions beneficial to the human body (hereinafter referred to as "functional ingredients") and that helps maintain health or prevent and / or improve certain diseases or symptoms, and there are no particular limitations on the form of the final product. For example, the functional health food may have a dosage form selected from the group consisting of powder, granules, tablets, capsules, pills, gels, jellies, suspensions, emulsions, syrups, tea bags, infused tea, and health drinks.

[0034] The content of the active ingredient (i.e., fraction of Syzygium formosum extract) contained in the health functional food can be appropriately selected depending on the form of the food, the desired use, etc., and is not particularly limited, and may be, for example, 0.1 to 50 wt % of the total food weight.

[0035] The health functional food may further contain at least one selected from the group consisting of various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents or natural flavoring agents, coloring agents, enhancers (cheese, chocolate, etc.), pectinic acid or its salts, alginic acid or its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc. The proportion of such additives is generally selected within the range of 0.001 to about 20 parts by weight per 100 parts by weight of the total health functional food, but is not limited thereto.

[0036] In yet another embodiment, there is provided a pharmaceutical composition for preventing or treating inflammatory skin diseases, comprising a fraction of a Syzygium formosum extract as an active ingredient.

[0037] Yet another embodiment provides a method for preventing or treating skin inflammation comprising administering a fraction of an extract of Syzygium formosum to an individual in need thereof.

[0038] The inflammatory skin disease may be selected from the group consisting of skin wounds, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, drug rash, and acne.

[0039] The term "prevention" as used herein refers to a method of partially or completely delaying or preventing the onset or recurrence of a disease, disorder, or its associated symptoms, preventing the acquisition or re-acquisition of a disease or disorder, or reducing the risk of acquiring a disease or disorder. For example, the prevention refers to any action of suppressing or delaying the onset of inflammation or inflammation-related diseases, disorders, or symptoms by administering a composition according to the present invention.

[0040] The term "amelioration" as used herein can refer to any action that at least reduces the parameters involved in the alleviation or treatment of a condition, such as the severity of symptoms.

[0041] The term "treatment" as used herein includes alleviation or amelioration of pathological symptoms, reduction in the area of ​​disease, delay or alleviation of disease progression, improvement, relief or stabilization of the disease state or symptoms, partial or complete recovery, prolongation of survival, and other beneficial results of treatment.

[0042] As used herein, the term "pharmaceutical composition" can refer to a molecule or compound that provides some beneficial effect when administered to a subject. Beneficial effects can include enabling a diagnostic determination; ameliorating a disease, symptom, disorder, or condition; reducing or preventing the onset of a disease, symptom, disorder, or condition; and generally addressing a disease, symptom, disorder, or condition.

[0043] The pharmaceutical composition may be administered parenterally in clinical settings and may be used in the form of a common pharmaceutical formulation. Parenteral administration may refer to administration via routes other than oral administration, such as rectal, intravenous, peritoneal, intramuscular, intraarterial, transdermal, nasal, inhalation, ocular, and subcutaneous. When the pharmaceutical composition of the present invention is used as a pharmaceutical, it may further contain one or more active ingredients exhibiting the same or similar functions.

[0044] The pharmaceutical compositions may be formulated in the form of solutions, suspensions, syrups, or emulsions in aqueous or oily media, or in the form of powders, powders, granules, tablets, or capsules, and may further contain dispersants or stabilizers for formulation. The pharmaceutical compositions may be formulated using commonly used diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Examples of suppository bases include Witepsol, Macrogol, Tween 61, cocoa butter, lauric butter, and glycerogelatin.

[0045] The pharmaceutical composition may be used by mixing with any of a variety of pharmaceutically acceptable carriers, such as physiological saline or organic solvents. To increase stability and water absorption, carbohydrates such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, or other stabilizers may be used as pharmaceuticals.

[0046] The pharmaceutically effective amount and effective dosage of the pharmaceutical composition may vary depending on the formulation method, administration method, administration time, and / or administration route of the pharmaceutical composition. The effective amount and effective dosage may vary depending on many factors, including the type and degree of response to be achieved by administration of the pharmaceutical composition, the type, age, weight, general health condition, symptoms and severity of disease, sex, diet, excretion, drugs used simultaneously or at different times in the individual, other components of the composition, and similar factors well known in the pharmaceutical arts. Those skilled in the art can easily determine and prescribe an effective dosage for the intended treatment. The pharmaceutical composition of the present invention may be administered once a day or in divided doses. Therefore, the dosage does not limit the scope of the present invention in any respect. The dosage of the pharmaceutical composition may be 1 μg / kg / day to 1,000 mg / kg / day.

[0047] The individual may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The individual may be in need of treatment for skin inflammation.

[0048] The terms and methods described for the above inventions are equally applicable between these inventions.

[0049] According to one embodiment, a composition containing a fraction of a Syzygium formosum extract as an active ingredient can be useful for preventing, improving, or treating inflammatory diseases by suppressing the expression of inflammatory cytokines, and has the effect of providing high stability to cosmetics containing the composition.

[0050] The present invention will be described in more detail below with reference to examples, but these are merely illustrative and are not intended to limit the scope of the present invention. It will be obvious to those skilled in the art that the examples described below can be modified within the scope of the essential gist of the invention. [Example]

[0051] Example 1 Preparation of Syzygium formosum extract Harvested and dried Syzygium formosum leaves were purchased from Nguyen Van Loung in Hanoi, Vietnam. 50 kg of dried Syzygium formosum leaves were added to 600 L of 70% (v / v) ethanol solution and extracted at 50°C for 24 hours, and then filtered to obtain a primary extract. The filtered Syzygium formosum was added to 600 L of 95% (v / v) ethanol solution and extracted at 50°C for 24 hours, and then filtered to obtain a secondary extract. The extract was concentrated and freeze-dried to obtain the final extract.

[0052] Example 2-1 Preparation of the first washing solution further purified with distilled water for the preparation of fractionated Syzygium formosum extract 2000 ml of distilled water was added to 100 g of the extract obtained in Example 1, and the mixture was mixed for 1 minute, and then centrifuged to recover the washings.

[0053] Example 2-2 Preparation of secondary cleaning solution by further purifying Syzygium formosum extract with distilled water 2000 ml of distilled water was added to the precipitate separated by centrifugation in Example 2-1, and the mixture was mixed for 1 minute, and then centrifuged to recover the washing liquid.

[0054] Example 2-3 Preparation of the third cleaning solution by further purifying Syzygium formosum extract with distilled water 2000 ml of distilled water was added to the precipitate separated by centrifugation in Example 2-2, and the mixture was mixed for 1 minute, and then centrifuged to recover the washing liquid.

[0055] Examples 2-4 Preparation of fractionated Syzygium formosum extract further purified with distilled water The precipitate separated by centrifugation in Example 2-3 was freeze-dried to obtain a fraction of Syzygium formosum extract containing high amounts of triterpenes.

[0056] Experimental Example 1 Confirmation of yield after additional purification with distilled water After further purification with distilled water, the yield was confirmed and the results are shown in Table 1.

[0057] [Table 1]

[0058] Experimental Example 2 Confirmation of changes in the content of active ingredients in fractions of Syzygium formosum extracts further purified with distilled water using LC-MS / MS (1)LC-MS / MS analysis conditions LC-MS / MS analysis was carried out under the conditions shown in Tables 2 and 3 below.

[0059] [Table 2]

[0060] [Table 3]

[0061] (2)Analysis results The nine active ingredients in the extract obtained in Example 1 and the purified liquid and fractions obtained in Examples 2-1 to 2-4 were quantified and shown in Table 4 below in ppm. It was confirmed that the fractions of the Syzygium formosum extract had a higher content of active ingredients than the Syzygium formosum extract.

[0062] [Table 4]

[0063] Experimental Example 3 Confirmation of cytotoxicity of Syzygium formosum extracts and fractions HaCaT cells, a keratinocyte cell line, were treated with the extracts and fractions obtained in Examples 1 and 2-4, respectively, and the cytotoxicity of each was examined using MTT solution after 24 hours.

[0064] Specifically, HaCaT cells were subcultured 6 to 11 times and seeded at 0.05 x 106 cells / well in 200 μL of medium (5% FBS, DMEM) in a 48-well plate and cultured overnight. The cultured cells were treated with various concentrations of Syzygium formosum extract and fractions and then cultured at 37°C for 24 hours. 10 μL of MTT solution (5 mg / mL in DPBS) was added and incubated for 3 hours. After removing the medium containing the MTT solution, 300 μL of DMSO was added and mixed for 5 minutes. 100 μL of the solution was pipetted into a 96-well plate and measured at 540 nm to calculate cell viability (%).

[0065] As a result, as shown in Figure 1, in the case of Syzygium formosum extract, a cell viability of over 80% was confirmed at a concentration of 50 μg / ml, and in the case of the fraction of Syzygium formosum extract, a cell viability of approximately 80% was confirmed at a concentration of 25 μg / ml.

[0066] Experimental Example 4 Comparison of the anti-inflammatory effects of Syzygium formosum extract and fractions The anti-inflammatory effect of fractions of Syzygium formosum extract was confirmed by examining the expression of inflammatory cytokines in response to the inflammatory response of cells caused by oxidative stress caused by UVB irradiation.

[0067] Specifically, HaCaT cells that had been passaged four times were seeded in 2 ml of medium at 0.5 x 106 cells / ml in 6-well plates and pretreated for 6 hours with Syzygium formosum extract and fractions at various concentrations. The positive control group was treated with vitamin C. After removing the medium, the cells were washed twice with DPBS. They were then irradiated with 20 mJ of UVB for 30 seconds in 500 μl of DPBS. The DPBS was then removed and 2 ml of DMEM serum-free medium containing 15 μg / ml of each extract was added, followed by incubation for 18 hours. RNA was then extracted and the expression of inflammatory cytokines was confirmed by polymerase chain reaction (PCR).

[0068] As a result, as shown in Figures 2 to 6, it was confirmed that the fractions of the Syzygium formosum extract inhibited the expression of inflammatory cytokines more effectively than vitamin C and the extract.

[0069] Experimental Example 5 Comparison of the active ingredient contents of fractionated Syzygium formosum extract and Centella asiatica extract The active ingredients of the fractions of the Syzygium formosum extract and the extracts of two species of Centella asiatica were compared.

[0070] Specifically, 40 ml of 70% ethanol was added to 2 g of dried Syzygium formosanum leaves to prepare extracts and fractions in the same manner as in Examples 1 and 2-1 to 2-4. 2 g of each of the two Centella asiatica species was added with 40 ml of 70% ethanol and extracted at 50°C for 24 hours, and the extract was collected. LC-MS / MS analysis was then performed under the conditions of Tables 2 and 3 to quantify the nine active ingredients, and the contents of the active ingredients are shown in Table 5 in ppm.

[0071] As a result, it was confirmed that the fraction of Syzygium formosum extract had a higher content of active ingredients than Centella asiatica extract.

[0072] [Table 5]

[0073] Experimental Example 6 Comparison of the anti-inflammatory effects of Centella asiatica extract, Syzygium formosum extract, and fractions The anti-inflammatory effect of fractions of Syzygium formosum extract was confirmed by examining the expression of inflammatory cytokines in response to the inflammatory response of cells caused by oxidative stress caused by UVB irradiation.

[0074] Specifically, HaCaT cells that had been passaged 16 times were seeded in 2 ml of medium at 0.5 x 106 cells / ml in 6-well plates and pretreated with Centella asiatica extract, Syzygium formosum extract, and fractions for 6 hours. After removing the medium, the cells were washed twice with DPBS. They were then exposed to 20 mJ of UVB for 30 seconds in 500 μl of DPBS. After removing the DPBS, 2 ml of DMEM serum-free medium containing 8.8 ppm of vitamin C, 0.43 ppm of Syzygium formosum extract, and 2.14 ppm of fractions were added and cultured for 18 hours. RNA was then extracted and the expression of inflammatory cytokines was confirmed by polymerase chain reaction (PCR).

[0075] As a result, as shown in Figures 7 to 10, it was confirmed that the fraction of Syzygium formosum extract inhibited the expression of inflammatory cytokines more effectively than Centella asiatica extract and Syzygium formosum extract. These results indicate that the fraction of Syzygium formosum extract has a stronger anti-inflammatory effect than Centella asiatica extract and Syzygium formosum extract.

[0076] Manufacturing Example 1 Preparation of cream containing Syzygium formosum extract In order to confirm the cosmetic material properties of the Syzygium formosum extract, creams were prepared in a conventional manner according to the composition shown in Table 5 below using the extract obtained in Example 1, and the content of the active ingredients in the extracts and fractions obtained in Examples 1 and 2-4 was varied during cream preparation.

[0077] [Table 6]

[0078] Manufacturing Example 2 Preparation of cream containing fractionated Syzygium formosum extract In order to confirm the cosmetic material properties of the fractions of Syzygium formosum extract, creams were prepared in a conventional manner according to the compositions shown in Table 6 below using the fractions obtained in Examples 2-4. The contents of the creams were varied depending on the difference in the contents of the active ingredients of the extracts and fractions obtained in Examples 1 and 2-4.

[0079] [Table 7]

[0080] Experimental Example 3 Confirmation of cosmetic material properties of fractions of Syzygium formosum extract through accelerated testing The cream prepared in Preparation Example 1 was stored at 45°C for 6 weeks, and the pH and color were measured every week. The results are shown in Table 7 and Figures 11 to 13. Figures 11 to 13 show the lightness (L), hue (a), and saturation (b) values ​​as a percentage (%) based on the values ​​measured at week 0. The viscosity measurement results are shown in Figure 14, with the red line indicating the viscosity of a typical cream.

[0081] As a result, as shown in Figures 11 to 14, the cream containing the fraction of Syzygium formosum extract maintained a viscosity about 5000 cP higher than the cream containing Syzygium formosum extract, and the lightness (L) was higher and the degree of change was smaller. In addition, it was confirmed that the changes in hue (a) and saturation (b) were smaller, confirming that the formulation was more stable.

[0082] [Table 8]

Claims

1. A method for producing a cosmetic composition for improving skin inflammation, which contains a fraction of an extract of Syzygium formosum as an active ingredient, the method comprising extracting Syzygium formosum with an aqueous ethanol solution, fractionating the extract with water to obtain a water fraction of the extract of Syzygium formosum, and then using the water fraction to produce the cosmetic composition.

2. A method for producing an external skin composition for improving skin inflammation, which contains a fraction of an extract of Syzygium formosum as an active ingredient, the method comprising extracting Syzygium formosum with an aqueous ethanol solution, fractionating the extract with water to obtain a water fraction of the extract of Syzygium formosum, and then producing the composition using the water fraction.

3. A method for producing a health functional food for improving skin inflammation, which contains a fraction of an extract of Syzygium formosum as an active ingredient, the method comprising extracting Syzygium formosum with an aqueous ethanol solution, fractionating the extract with water to obtain a water fraction of the extract of Syzygium formosum, and then using the water fraction to produce the health functional food.

4. A method for producing a pharmaceutical composition for preventing or treating inflammatory skin diseases, comprising a fraction of an extract of Syzygium formosum, the method comprising extracting Syzygium formosum with an aqueous ethanol solution, fractionating the extract with water to obtain a water fraction of an extract of Syzygium formosum, and then using the water fraction to produce the pharmaceutical composition.

5. The method according to any one of claims 1 to 4, wherein the water fraction is obtained by repeating fractionation one to five times.

6. 5. The method according to claim 1, wherein the water fraction contains 11,400 ppm of ursolic acid and 4,100 ppm of oleanolic acid.

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