Composition for alleviating atopic dermatitis using vaccinium bracteatum extract

Abstract

The present invention discloses a composition for alleviating atopic dermatitis using a Vaccinium bracteatum extract, which exhibits the effects of not only significantly lowering the severity clinical index, which is a comprehensive index of atopic dermatitis symptoms, and alleviating itchiness, which is the most representative symptom of atopic dermatitis, but also significantly reducing the size of immune-related tissues such as those in the spleen and lymph nodes, and decreasing the blood concentration of IgE, which is the most representative antibody for atopic diseases.

Description

Composition for improving atopic dermatitis using extract of *Mulberry*

[0001] The present invention relates to a composition for improving atopic dermatitis using an extract of Vaccinium bracteatum.

[0002] Atopic dermatitis is a recurrent chronic skin disease that commonly occurs in infants and young children, but can persist into adulthood or develop anew in adulthood, and its prevalence has recently been on the rise (Kor J Pharmacogn., 43:59-65, 2012).

[0003] Although the cause of atopic dermatitis has not yet been clearly identified, immunological factors have been reported, including an abnormal increase in IgE, a trigger for allergic reactions; a decrease in the number and dysfunction of T cells, which play a central role in cellular immunity; the infiltration of monocytes and macrophages; an increase in the number of mast cells and eosinophils; an increase in the number of CD4+ T lymphocytes; and Th1 / Th2 imbalance resulting from an increase in the number of Th2 cells compared to Th1 cells (J Invest Dermatol., 96:523-526, 1991; J Invest Dermatol., 97:389-394, 1991; Immunol., 11:81-88, 1999; Curr Drug Targets Inflamm Allergy., 2:199-120, 2003; J Allergy Clin Immunol., 107:871-877, 2001; Adv Immunol., 78:57, 2001; International Immunology, 14(7):767-773, 2002; Pediatr Allergy Immunol., 19:605-613, 2008).

[0004] Atopic dermatitis is a type of allergic disease characterized by dry skin, skin itching (pruritus), and inflammation, and exhibiting symptoms such as perifollicular hypertrophy, cheilitis, lichenification, and eczematous lesions (Korean J Invest Dermatol., 14:67-72, 2007). In particular, itching is considered important in the treatment of atopic dermatitis because scratching leads to secondary worsening of skin symptoms, such as wounds and secondary infections.

[0005] In experiments on atopic dermatitis, researchers not only verify the production of immune substances in the blood but also confirm direct skin symptoms using animal models such as skin irritation models, diet-induced models, environmental-induced models, and ovalbumin skin sensitization models. Generally, ovalbumin skin sensitization models and skin irritation models are widely used; however, skin irritation models tend to be preferred because the ovalbumin skin sensitization model takes a relatively longer time to induce compared to chemical skin irritation models, and actual dermatitis symptoms are difficult to observe visually. The skin irritation model is a method of inducing atopic dermatitis using chemical substances (DNCB: dintrochlorobenzene, DNFB: dinitrofluorobenzene, PiCl: picryl chloride). As an acute atopic induction model, it can induce dermatitis within a relatively short period and is useful for research due to the advantage of being able to visually observe lichen and blisters upon induction.

[0006] Currently, steroids that inhibit inflammatory responses and cytokine production are primarily used to treat atopic dermatitis; however, long-term administration can lead to various side effects, such as skin atrophy or growth retardation. Consequently, the use of non-steroidal anti-inflammatory drugs (NSAIDs) has been increasing recently. Nevertheless, NSAIDs also carry various side effects, including symptoms such as erythema, itching, edema, chafing, and lichenification, as well as weakened immunity, making it difficult to achieve a fundamental cure for atopic dermatitis (Arellano FM et al., J Invest Dermatol. 2007 Apr;127(4):808-16.). Therefore, research is needed to identify substances with excellent therapeutic effects from natural products that are relatively safe.

[0007] The objective of the present invention is to provide a composition for improving atopic dermatitis using an extract of *Mulberry*.

[0008] Other objects or specific objectives of the present invention will be presented below.

[0009] As confirmed in the following examples and experimental examples, the present invention was completed by confirming that when the fruit extract and leaf extract of *Musaengnamu* were orally administered to a mouse animal model in which atopic dermatitis was induced by DNCB (dintrochlorobenzene), they significantly lowered the clinical severity index, which is a comprehensive indicator of symptoms of atopic dermatitis such as itching, erythema, hemorrhage, edema, excoriation, erosion, scarring, and dryness, and not only alleviated itching, which is the most representative symptom of atopic dermatitis, but also significantly reduced the size of immune-related tissues such as lymph nodes and lowered the blood concentration of IgE, the most representative antibody of atopic diseases.

[0010] The present invention is provided based on these experimental results and can be understood in one aspect as a composition for improving atopic dermatitis comprising a *Ilex crenata* extract as an active ingredient, in another aspect as a composition for suppressing skin hypersensitivity reactions comprising a *Ilex crenata* extract as an active ingredient, and in yet another aspect as a composition for improving itching (pruritus) comprising a *Ilex crenata* extract as an active ingredient. Furthermore, the present invention can also be understood as a composition for suppressing immune responses comprising a *Ilex crenata* extract as an active ingredient.

[0011] In this specification, "Mulberry tree extract" refers to an extract obtained by leaching the stem, branch, leaf, fruit, root, flower, etc. of a Mulberry tree to be extracted using water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixture thereof, an extract obtained using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract. The extraction method may be any method such as cold maceration, reflux, heating, ultrasonic radiation, or supercritical extraction, taking into consideration the polarity of the active substance, the degree of extraction, and the degree of preservation. The term "fractionated extract" includes fractions obtained by suspending an extract in a specific solvent and then mixing and settling it with a solvent of different polarity, as well as fractions obtained by adsorbing the crude extract onto a column packed with silica gel or the like and using a hydrophobic solvent, a hydrophilic solvent, or a mixture thereof as the mobile phase. Furthermore, the term "extract" includes concentrated liquid or solid extracts from which the extraction solvent has been removed by methods such as freeze-drying, vacuum drying, hot-air drying, or spray drying. Preferably, it refers to an extract obtained using water (including hot water), ethanol, or a mixture thereof as the extraction solvent.

[0012] In addition, the term "active ingredient" in this specification means an ingredient that exhibits the intended activity alone or can exhibit activity together with a carrier that is inactive itself.

[0013] Furthermore, in this specification, "atopic dermatitis" is defined to include all diseases classified as atopic dermatitis in the art, regardless of the direct or indirect cause of their occurrence. Typically, atopic dermatitis is classified into infantile atopic dermatitis, pediatric atopic dermatitis, adult atopic dermatitis, and pregnant woman atopic dermatitis depending on the time of onset or the subject of the invention; in this specification, atopic dermatitis includes all of these types of atopic dermatitis.

[0014] In addition, in this specification, "improvement" includes the treatment, prevention, and alleviation of symptoms of atopic dermatitis, skin itching, etc.

[0015] The composition of the present invention may include the active ingredient in any amount (effective amount) as long as it can exhibit improvement activity, such as that for atopic dermatitis intended for treatment, depending on the use, formulation, and purpose of combination. A typical effective amount will be determined within the range of 0.001 weight % to 15 weight % based on the total weight of the composition. Here, "effective amount" refers to the amount of the active ingredient included in the composition of the present invention that can exhibit intended medical and pharmacological effects, such as improvement of atopic dermatitis, when the composition of the present invention is administered to mammals, preferably humans, the subjects of application, for a period of administration recommended by medical professionals. Such an effective amount may be determined experimentally within the ordinary capacity of a person skilled in the art.

[0016] In addition to the active ingredient, the composition of the present invention may further include any compound or natural extract known in the art to have the said activity and whose safety has already been verified, for the purpose of enhancing or reinforcing the activity of improving atopic dermatitis or itching, or the activity of suppressing immune responses, or for the purpose of improving the convenience of taking, ingesting, or using by adding similar activities such as skin whitening activity, skin wrinkle improvement activity, or skin protective activity (inhibition of skin damage caused by ultraviolet rays, skin moisturization, etc.).

[0017] These compounds or extracts include compounds or extracts listed in pharmacopoeias of various countries (the "Korean Pharmacopoeia" in Korea), health functional food codes of various countries (the "Standards and Specifications for Health Functional Foods" notified by the Ministry of Food and Drug Safety in Korea), and functional cosmetics codes of various countries (the "Standards and Test Methods for Functional Cosmetics" notified by the Ministry of Food and Drug Safety in Korea); compounds or extracts that have received product approval in accordance with the laws of various countries governing the manufacture and sale of pharmaceuticals (the "Pharmaceutical Affairs Act" in Korea); compounds or extracts whose functionality has been recognized in accordance with the laws of various countries governing the manufacture and sale of health functional foods (the "Health Functional Foods Act" in Korea); and compounds or extracts whose functionality has been recognized in accordance with the laws of various countries governing the manufacture and sale of functional cosmetics (the "Cosmetics Act" in Korea).

[0018] These ingredients include, for example, a complex of heat-treated dried powder of Enterococcus faecalis and guava leaf extract, which are recognized for their functionality of 'alleviating hypersensitive immune responses' under the 'Act on Health Functional Foods'; a complex of kiwi extract, Perilla leaf extract, and Picao Preto powder; PLAG (1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol); and L., which is recognized for its functionality of 'improving hypersensitive skin conditions'. sakeiProbio 65, gamma-linolenic acid-containing oil, fruit and vegetable-derived lactic acid bacteria (L. plantarum CJLP133), probiotics ATP, etc., and arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, oil-soluble licorice (Glycyrrhiza) extract, etc., recognized as skin whitening ingredients in the Cosmetic Code under the Korean Cosmetic Act (Notification of the Ministry of Food and Drug Safety, "Standards and Test Methods for Functional Cosmetics"), and also retinol, retinyl palmitate, adenosine, polyethoxylated amide, etc., recognized as skin wrinkle improvement ingredients in the same Korean Cosmetic Code, and also drometrizole, drometrizole trisiloxane, digalloyl trioleate, dimethicodiethylbenzalmalonate, diethylhexyl butamidotriazone, etc., recognized as UV protection ingredients in the same Korean Cosmetic Code, and Korea Examples include complexes such as pine bark extract recognized as a UV protection ingredient under the "Act on Health Functional Foods," phosphatidylserine, fingerroot extract powder, and complex extracts such as red ginseng and Cornus fruit. One or more of these compounds or natural extracts may be included in the composition of the present invention together with the active ingredients.

[0019] The composition of the present invention can be understood as a food composition in specific embodiments.

[0020] The food composition of the present invention can be manufactured in any form, for example, as beverages such as tea, juice, carbonated drinks, and isotonic drinks; processed dairy products such as milk and yogurt; chewing gums; rice cakes, Korean confectionery, bread, cookies, and noodles; and health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and bars.

[0021] In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification, as long as it complies with the regulations in effect at the time of manufacture and distribution. For example, it may be a health functional food under the Korean "Act on Health Functional Foods," or a confectionery, legume, tea, beverage, special dietary food, etc., according to each food type in the Food Code of the Korean "Food Sanitation Act" (Notification of the Ministry of Food and Drug Safety "Standards and Specifications for Food").

[0022] The food composition of the present invention may include food additives in addition to its active ingredients. Food additives can generally be understood as substances added to, mixed with, or permeated into food during the manufacture, processing, or preservation of food; since they are consumed daily and over a long period of time along with food, their safety must be guaranteed. Food additive codes in accordance with national laws governing the manufacture and distribution of food (the "Food Sanitation Act" in Korea) restrictively define food additives with guaranteed safety in terms of composition or function. The Korean Food Additive Code (Notification of the Ministry of Food and Drug Safety, "Standards and Specifications for Food Additives") classifies food additives into chemically synthesized products, natural additives, and mixed preparations in terms of composition, and these food additives are classified into sweeteners, flavorings, preservatives, emulsifiers, acidulants, thickeners, etc., in terms of function.

[0023] Sweeteners are used to impart a suitable sweetness to food, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, natural sweeteners are used, and examples of natural sweeteners include corn syrup solids, honey, sucrose, fructose, lactose, maltose, etc.

[0024] Flavoring agents can be used to improve taste or aroma, and both natural and synthetic ones may be used. Preferably, natural ones are used. When natural ones are used, nutritional enhancement can be achieved in addition to flavor. Natural flavoring agents may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, etc., or from green tea leaves, Solomon's seal, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. Additionally, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc., may be used. Natural flavoring agents may be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, etc.

[0025] Calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. may be used as preservatives, and acacia gum, carboxymethylcellulose, xanthan gum, pectin, etc. may be used as emulsifiers, and acetic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. may be used as acidifiers. Acidifiers may be added to the food composition to achieve an appropriate acidity, in addition to for the purpose of enhancing flavor, for the purpose of inhibiting the growth of microorganisms.

[0026] As thickening agents, suspending agents, settling agents, gel-forming agents, puffing agents, etc. may be used.

[0027] In addition to the food additives described above, the food composition of the present invention may include physiologically active substances or minerals known in the art and whose safety as food additives is guaranteed, for the purpose of supplementing and reinforcing functionality and nutritional value.

[0028] Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, etc., and examples of minerals include calcium preparations such as calcium citrate, magnesium preparations such as magnesium stearate, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc.

[0029] The food composition of the present invention may include the food additives described above in an appropriate amount to achieve the purpose of their addition, depending on the product type.

[0030] Regarding other food additives that may be included in the food composition of the present invention, reference may be made to the food codes or food additive codes of each country.

[0031] The composition of the present invention may be understood as a pharmaceutical composition in other specific embodiments.

[0032] The pharmaceutical composition of the present invention may be prepared as an oral or parenteral formulation according to the route of administration by conventional methods known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The route of administration may be any suitable route including a local route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissues, and may also be used in combination of two or more routes. An example of a combination of two or more routes is a case where two or more formulations of drugs according to the route of administration are combined, for example, one drug is administered first via the intravenous route and another drug is administered second via the local route.

[0033] Pharmaceutically acceptable carriers are known in the art according to the route of administration or dosage form, and specifically, one may refer to the pharmacopoeias of each country, including the "Korean Pharmacopoeia."

[0034] When the pharmaceutical composition of the present invention is prepared as an oral formulation, it may be prepared in the form of powder, granules, tablets, pills, coated tablets, capsules, liquids, gels, syrups, suspensions, wafers, etc., in accordance with methods known in the art together with a suitable carrier. Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol; starches such as corn starch, potato starch, and wheat starch; celluloses such as cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; polyvinylpyrrolidone; water; methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate; mineral oil; malt; gelatin; talc; polyols; vegetable oils; ethanol; glycerol, etc. In the case of formulation, appropriate binders, lubricants, disintegrants, coloring agents, diluents, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferrite, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweeteners, sodium alginate, polyethylene glycol, wax, etc. Suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium and calcium salts, polyethylene glycol, etc. Examples of disintegrants include starch, methylcellulose, agar, bentonite, xanthan gum, starch, alginic acid, or its sodium salt, etc. In addition, diluents include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc.

[0035] When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it may be formulated in the form of an injectable, transdermal, nasal inhalant, and suppository according to methods known in the art with a suitable carrier. When formulated as an injectable, an aqueous isotonic solution or suspension may be used as a suitable carrier; specifically, isotonic solutions such as PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or 5% dextrose may be used. When formulated as a transdermal formulation, it may be formulated in the form of an ointment, cream, lotion, gel, topical solution, paste, liniment, aerosol, etc. In the case of nasal inhalers, they can be formulated in the form of an aerosol spray using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide, and when formulated as suppositories, the carriers may include Witepsol, Tween 61, polyethylene glycols, cocoa starch, laurin starch, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, and sorbitan fatty acid esters.

[0036] Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to literature such as [Remington's Pharmaceutical Sciences (19th ed., 1995)]. The said literature is to be considered part of this specification.

[0037] A preferred dosage of the pharmaceutical composition of the present invention may be in the range of 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g / kg, depending on the patient's condition, body weight, gender, age, severity of the patient, and route of administration. Administration may be administered once a day or divided into several doses. Such dosages shall not be construed as limiting the scope of the present invention in any aspect.

[0038] The composition of the present invention can be understood as a cosmetic composition in another specific embodiment.

[0039] Even if the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may take on any product classification in terms of its use or legal classification, and specifically, it may be a functional cosmetic or a non-functional general cosmetic used for purposes such as improving skin troubles or atopic dermatitis. In terms of product form, it may also take on any product form, specifically, it may take the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, spray, etc. In terms of specific product form, it may be a formulation such as a softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder.

[0040] The cosmetic composition of the present invention may include, in addition to the active ingredient, ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, colorants, and fragrances, and carriers.

[0041] In the case where the formulation of the present invention is a paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0042] In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0043] When the formulation of the present invention is a solution or an emulsion, a solvent, a solubilizing agent, or an emulsifying agent is used as a carrier component, specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, etc. may be used.

[0044] In the case where the formulation of the present invention is a suspension, liquid diluents such as water, ethanol, or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, etc. may be used as carrier components.

[0045] In the case where 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, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester, etc. may be used as a carrier component.

[0046] The cosmetic composition of the present invention can be prepared according to the method of preparing cosmetic compositions commonly practiced in the art, except that it includes an active ingredient that exhibits activity in improving atopic dermatitis.

[0047] As described above, according to the present invention, a composition for improving atopic dermatitis and a composition for improving itching can be provided using an extract of *Mulberry*.

[0048] The composition of the present invention can be commercialized into products such as food, cosmetics, and pharmaceuticals for purposes such as improving atopic dermatitis and alleviating itching.

[0049] Figure 1 is a schematic diagram related to an animal model experiment in which atopic dermatitis was induced with DNCB.

[0050] Figure 2 shows the results of measuring the atopic severity index in an animal model of atopic dermatitis.

[0051] Figure 3 shows the results of evaluating the degree of itching in an animal model of atopic dermatitis.

[0052] Figure 4 shows the results of lymph node size measurement in an animal model of atopic dermatitis.

[0053] Figure 5 shows the results of evaluating the degree of antibody production in an animal model of atopic dermatitis.

[0054] The present invention will be described below with reference to examples. However, the scope of the present invention is not limited to these examples.

[0055]

[0056] <Example> Evaluation of the activity of *Mulberry* extract in improving atopic dermatitis, etc.

[0057] 1. Preparation of extract

[0058] 50 L of distilled water was added to 5 kg of dried Vaccinium bracteatum fruit, and the mixture was heated to 100°C for 3 hours for extraction. The extract was filtered and concentrated under reduced pressure. The concentrated hot water extract was freeze-dried using a freeze dryer at -40°C to 40°C for 72 hours. By the above method, 530 g (10.6%) of Vaccinium bracteatum fruit extract (VBF) was obtained.

[0059] Vaccinium bracteatum leaf extract (VBL) was also obtained in an amount of 1382g (27.6%) using the same method as above.

[0060] 2. Experimental Method

[0061] 2.1 Experimental Animals and Experimental Methods

[0062] To experimentally confirm the effect of the extract of *Mustard rhizome* on improving symptoms of atopic dermatitis, NC / Nga mice (Vestergaard C, Yoneyama H, Murai M, Nakamura K, Tamaki K, Terashima Y, Imai T, Yoshie O, Irimura T, Mizutani H and Matsushima K. Overproduction of Th2-specific chemokines in NC / Nga mice exhibiting atopic dermatitis-like lesions, J Clin Invest 104(8),1097-105; 1999), an animal model of atopic dermatitis known in the industry, were used. To evaluate the efficacy of Vaccinium bracteatum extract in improving atopic dermatitis and immunomodulating atopic dermatitis in an animal model of atopic dermatitis, mice were divided into a naive group and an atopic dermatitis-induced group. The atopic dermatitis-induced group was further divided into a group treated with HDM (house dust mite) / DNCB (dinitrochlorobenzene) (negative control group), a group treated with HDM / DNCB and oral administration of Vaccinium bracteatum fruit extract (VBF) at 100 mg / kg, a group treated with Vaccinium bracteatum leaf extract (VBL) at 100 mg / kg, and a group treated with HDM / DNCB and CJLP133 (CJ Cheiljedang Co. Seoul, Korea, 800 mg / kg) (positive control group). The positive control (CJLP-133 800 mg / kg) is a health functional food ingredient individually recognized by the Ministry of Food and Drug Safety for its function of inhibiting skin hypersensitivity reactions.

[0063] Atopic dermatitis was induced primarily in NC / Nga mice with depilated backs by applying 1% DNCB, and a secondarily induced one week later by applying 0.5% DNCB and 90 mg of HDM ointment to the depilated area. Subsequently, atopic dermatitis was induced and sustained tertiarily by applying 0.5% DNCB every other day for three weeks. Simultaneously, for three weeks, CJLP133 and rhododendron fruit extract (100 mg / kg) were orally administered to the positive control group daily, and rhododendron leaf extract (100 mg / kg) was applied topically. Body weight, clinical symptoms, and scratching were measured weekly, and after the experiment was completed, the mice were sacrificed to measure spleen weight, lymph node weight, etc.

[0064] 2.2 Evaluation of Impact on Clinical Symptoms

[0065] The above clinical analysis was evaluated by scoring it as shown in Table 1 below, divided into weekly skin clinical analysis, itching, erythema and hemorrhage, edema, wounds and dryness of the ear area, etc., to assess the efficacy of the extract of *Mulberry* in improving atopic dermatitis and immune modulating in an animal model of atopic dermatitis.

[0066] Point 1 2 3 1 Itching (Mild Moderate Severe) 2 Erythema / Hemorrhage (Mild Moderate Severe) 3 Edema (Mild Moderate Severe) 4 Excoriation / Erosion (Mild Moderate Severe) 5 Scarring / Dryness (Mild Moderate Severe)

[0067] 2.3 Evaluation of the effect on improving skin itching symptoms Changes in skin itching symptoms, a representative clinical symptom of atopic dermatitis, were evaluated. To compare and observe itching symptoms before and after inducing atopic dermatitis, and after orally administering CJLP133 to the positive control group and *Mulberry tree* extract (100 mg / kg), the behavior of mice was filmed for 1 hour under dark conditions every week, and the number of scratches was measured by observing the footage.

[0068] 2.4 Evaluation of the Impact on Changes in Immune-Related Tissue Size

[0069] Atopic dermatitis induces various reactions within immune organs, primarily causing hypertrophy of these organs. Therefore, to observe the effects of *Musaengnamu* extract on immune-related organs, the weight of the spleen and lymph nodes was measured after the experiment was completed by extracting the spleen and lymph nodes.

[0070] 2.6 Evaluation of the Effect on the Production of Immunity-Related Antibodies

[0071] Overexpression of IgE in serum is known to be the most representative marker of atopic diseases. Therefore, to evaluate the effect of *Mulberry* extract on IgE production in animals with induced atopic dermatitis, the animals were sacrificed and serum was isolated to quantify the production of IgE, IgG1, and IgG2a using an ELISA kit (Bethyl Laboratories, Inc).

[0072] 3. Experimental Results

[0073] 3.1 Results of Evaluation of Clinical Symptoms

[0074] The results are shown in Figure 2. Referring to Figure 2, it can be seen that the fruit and leaf extracts (100 mg / kg) of the *Musaengnamu* tree alleviated atopic-related clinical symptoms significantly compared to the atopic dermatitis-induced group (DNCB group) at weeks 3–4, while showing effects similar to the positive control group (CJLP-133 800 mg / kg).

[0075] 3.2 Evaluation Results of Itching Symptoms

[0076] The results are shown in Figure 3. Both the fruit and leaf extracts (100 mg / kg) significantly relieved skin itching symptoms compared to the atopic dermatitis-induced group (DNCB group) starting from week 3 to 4, and showed effects similar to the positive control group (CJLP-133 800 mg / kg).

[0077] 3.3 Results of Lymph Node Size Assessment

[0078] The results are shown in Figure 4, where the fruit and leaf extracts (100 mg / kg) of the *Musaengnamu* tree both significantly reduced the size of lymph nodes compared to the atopic dermatitis-induced group (DNCB group).

[0079] In Figure 6, aLN, bLN, cNL, and dNL represent the axillary lymph node, brachial lymph node, superficial cervicals lymph node, and inguinal lymph node, respectively.

[0080] 3.4 Evaluation Results of Antibody Production Level

[0081] The results are shown in Figure 5, where the fruit and leaf extracts (100 mg / kg) of the Japanese quince tree both showed the effect of lowering the blood concentrations of IgE, IgG1, and IgG2a compared to the atopic dermatitis-induced group (DNCB group).

Claims

1. A composition for improving atopic dermatitis containing a *Morus alba* extract as an active ingredient.

2. In Paragraph 1, The above extract is a composition that is a heat or leaf extract of the Japanese zelkova tree.

3. In Paragraph 1, The above extract is a composition in which the extract is a water, ethanol, or mixed solvent extract thereof.

4. In any one of paragraphs 1 through 3, The above composition is a food composition.

5. In any one of paragraphs 1 through 3, The above composition is a pharmaceutical composition.

6. A food composition for inhibiting skin hypersensitivity reactions containing *Morus alba* extract as an active ingredient.

7. A food composition for improving itching containing an extract of *Morus alba* as an active ingredient.

8. A food composition for suppressing immune responses containing an extract of *Morus alba* as an active ingredient.

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