Composition for preventing, alleviating, or treating atopic dermatitis, containing extract of scutellaria baicalensis and raphanus sativus l. seeds as active ingredient

Abstract

The present invention relates to a composition for preventing, alleviating, or treating atopic dermatitis, containing an extract of Scutellaria baicalensis and Raphanus sativus L. seeds as an active ingredient. Specifically, a Scutellaria baicalensis extract and a Raphanus sativus L. seed extract exhibit cytotoxicity or have no significant difference in NO production inhibitory effects, whereas the extract of Scutellaria baicalensis and Raphanus sativus L. seeds of the present invention, extracted after mixing at a weight ratio of 2-4:8-6, significantly inhibits LPS-induced NO production without cytotoxicity, reduces the thickness of atopic dermatitis-induced skin in an animal model, has a skin moisturizing effect, and significantly reduces the production of atopic dermatitis-related factors of IgE and IL-4, and thus the present invention can be usefully used in health functional foods, medicines and the like for preventing, alleviating, or treating atopic dermatitis.

Description

Composition for preventing, improving or treating atopic dermatitis containing extracts of gold and Nabokja as active ingredients

[0001] The present invention relates to a composition for preventing, improving or treating atopic dermatitis containing extracts of gold and Nabokja as active ingredients.

[0002]

[0003] This work was supported by the Ministry of Science and ICT's major projects (Project Numbers: 1711195904 and 1711177839).

[0004] Atopic dermatitis is a chronic allergic disease caused by genetic, environmental, and immunological factors. It manifests as abnormalities in the stratum corneum, the outermost layer of the skin that acts as a protective barrier. It is exacerbated in dry climates. Atopic dermatitis has a global prevalence of approximately 10-20%, most commonly affecting infants between one month and one year of age. While the condition typically improves somewhat by age three, it can recur depending on environmental factors and individual differences.

[0005] The main symptoms of atopic dermatitis include severe itching, dry skin, rash, oozing, crusting, and scaly skin (scales), often accompanied by chronic skin inflammation. The itching, which triggers the urge to scratch, is induced by itch mediators such as histamine. While scratching can easily alleviate symptoms in healthy individuals, in atopic patients, the more they scratch, the more they feel the urge to scratch, leading to even more intense scratching. Scratching in atopic patients disrupts the skin barrier, leading to secondary infections and further aggravation of inflammation. While atopic dermatitis may temporarily improve, it can recur and worsen due to environmental and dietary irritants, leading to repeated periods of exacerbation and relief.

[0006] The cause of atopic dermatitis is not well known, but it has been revealed that it is mainly related to genetic factors and immune response. In addition, it seems to be caused by a complex interaction of dry skin, the characteristic of feeling skin itching more easily than normal people, infection by bacteria, viruses, and fungi, emotional factors, and environmental factors. Inflammation accompanying atopic dermatitis is caused by excessive immune cell activity, resulting in the production of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α) and interleukin-1 (IL-1), as well as hydroxyl radicals (OH), superoxide radicals (O2). - ), and hydrogen peroxide (H2O2), which cause damage to surrounding tissues because they produce a large amount of reactive oxygen species (ROS).

[0007] Prescriptions for atopic dermatitis primarily include medications such as steroids, antihistamines, and antibiotics. While steroids (corticosteroids) have anti-inflammatory and immunosuppressive effects and are highly effective, long-term use can lead to side effects such as skin weakening, systemic hormonal symptoms, and addiction. Recent research on the treatment of atopic dermatitis focuses on immunosuppressants and newer antihistamines. However, antihistamines alone do not completely block allergic reactions. This is because histamine is not the only chemical messenger that triggers allergic reactions. Therefore, there is a pressing need to develop novel atopic dermatitis treatment compositions utilizing natural products that are effective against atopic dermatitis while minimizing side effects.

[0008] Meanwhile, Scutellaria baicalensis refers to the perennial herb Scutellaria baicalensis, which belongs to the Labiatae family. It can specifically refer to the dried root of Scutellaria baicalensis, but is not limited to the root, stem, bud, flower, or seed of Scutellaria baicalensis. Scutellaria baicalensis has the effects of dehumidifying heat and clotting, and has been used to treat hypertension and epidemic meningitis. Its pharmacological effects that have been discovered so far include antibacterial, anti-inflammatory, anti-allergic, diuretic, hyperlipidemia-improving, and intestinal motility-suppressing effects.

[0009] Nabokja is the ripened seed of the radish (Raphanus sativus L.), a plant belonging to the Cruciferae family. The dried seeds are oval or round, slightly flattened, and approximately 3 mm long and 2.5 mm wide. The surface is reddish brown, with several vertical grooves on one side and an ovule at one end. The ovule is brown and protrudes like a round dot. When observed under a magnifying glass, the entire seed has a dense mesh pattern. The texture is firm, and when split, it reveals yellowish-white or yellowish kernels containing essential oil. When the seeds ripen in summer and fall, the seed heads are harvested and dried in the sun. The seeds are then rubbed by hand to remove impurities and dried in the sun before being used as a herbal medicine. Nabokja contains antibacterial substances, exhibiting a marked inhibitory effect against Staphylococcus aureus and Escherichia coli, and possesses antifungal properties against skin fungi such as Trichophyton erythematosus and Trichophyton erythematosus. And it has the effect of treating symptoms of rising energy, calming breathing, aiding digestion, and removing phlegm, and is prescribed to treat bronchial asthma caused by seawater, stagnation of energy due to food accumulation, chest tightness and heaviness, and second-degree fever caused by dysentery. Recently, it has been reported that extracts of Nabokja or some of its components exhibit potent antimutagenic, antibacterial, antihypertensive, and anti-inflammatory effects.

[0010] As for prior art related to atopic dermatitis, Korean Patent No. 0761248 discloses a composition for treating atopic dermatitis containing bamboo and gold extracts as active ingredients, and Korean Patent Publication No. 2011-0040024 discloses a composition for treating atopic dermatitis containing bamboo and gold extracts as active ingredients, and ... TM ) and an atopic dermatitis improvement cosmetic composition containing the same are disclosed. However, the 'composition for preventing, improving or treating atopic dermatitis containing the extracts of Golden and Nabokja as active ingredients' of the present invention has not yet been disclosed.

[0011] The present invention was derived from the above needs, and the present invention provides a composition for preventing, improving, or treating atopic dermatitis, which contains extracts of Scutellaria Baicalensis and Nabokja as active ingredients, and more specifically, while the extracts of Scutellaria Baicalensis and Nabokja did not show cytotoxicity or significantly differ in the effect of inhibiting NO production, the extracts of Scutellaria Baicalensis and Nabokja mixed and extracted at a weight ratio of 2-4:8-6 of the present invention significantly inhibited NO production induced by LPS without cytotoxicity, reduced the thickness of skin induced with atopic dermatitis in an animal model, had a skin moisturizing effect, and significantly reduced the production of IgE and IL-4, which are factors related to atopic dermatitis, thereby completing the present invention.

[0012] In order to solve the above problem, the present invention provides a health functional food composition for preventing or improving atopic dermatitis, which contains extracts of gold and Nabokja as active ingredients.

[0013] In addition, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis, which contains extracts of golden and Nabokja as active ingredients.

[0014] In addition, the present invention provides an over-the-counter drug for preventing or improving atopic dermatitis, which contains extracts of gold and Nabokja as active ingredients.

[0015] In addition, the present invention provides a feed additive for preventing or improving atopic dermatitis, which contains extracts of gold and Nabokja as active ingredients.

[0016] In addition, the present invention provides a veterinary composition for preventing or treating atopic dermatitis, comprising extracts of gold and Nabokja as active ingredients.

[0017] The present invention relates to a composition for preventing, improving or treating atopic dermatitis, comprising extracts of Scutellaria baicalensis and Nabokja jasminoides as active ingredients. Specifically, while the extracts of Scutellaria baicalensis and Nabokja jasminoides show no significant difference in cytotoxicity or NO production inhibition effect, the extracts of Scutellaria baicalensis and Nabokja jasminoides extracted by mixing them in a weight ratio of 2-4:8-6 of the present invention significantly inhibit LPS-induced NO production without cytotoxicity, reduce the thickness of skin induced with atopic dermatitis in an animal model, have a skin moisturizing effect, and significantly reduce the production of IgE and IL-4, which are factors related to atopic dermatitis.

[0018] Figure 1 shows the results of confirming the cell viability of extracts of gold and rhododendron amurense prepared according to the weight ratio of gold and rhododendron amurense of the present invention. Con represents the control group that was not treated with anything.

[0019] Figure 2 shows the results of confirming the amount of NO (Nitric Oxide) produced by extracts of Scutellaria Baicalensis and Nabokja prepared according to the weight ratio of Scutellaria Baicalensis and Nabokja in RAW 264.7 cells. # indicates that NO production in the LPS-treated group significantly increased compared to the untreated control group (Con), p<0.05, and * indicates that NO production in the extracts of Scutellaria Baicalensis and Nabokja of the present invention mixed and extracted at weight ratios of 4:6, 3:7, and 2:8 significantly decreased compared to the LPS-treated group, p<0.05.

[0020] Figure 3 shows the results of comparing the NO production (A) and cell viability (B) of the Scutellaria Baicalensis and Nabokja extracts mixed and extracted at a weight ratio of 4:6 of the present invention and the Scutellaria Baicalensis extract and Nabokja extract alone. Hwang40 means 40㎍ / ㎖ of Scutellaria Baicalensis extract, Na60 means 60㎍ / ㎖ of Nabokja extract, and Hwang40 / Na60 means an extract mixed and extracted at a weight ratio of 4:6 of Scutellaria Baicalensis. ### indicates that NO production in the LPS-treated group significantly increased compared to the untreated control group (Con), or that the cell viability of the Scutellaria Baicalensis extract (Hwang40) significantly decreased compared to the untreated control group (Con), and p<0.001, and ** indicates that NO production in the Scutellaria Baicalensis extract (Hwang40) and the Scutellaria Baicalensis and Nabokja extracts (Hwang40 / Na60) mixed and extracted at a weight ratio of 4:6 of the present invention significantly decreased compared to the LPS-treated group, and p<0.01.

[0021] Figure 4 shows the results of confirming the differentiation inhibitory effect of the goldenrod and Nabokja extracts mixed and extracted at a weight ratio of 4:6 of the present invention on CD4+ T cells. A shows the degree of CD4+ T cell differentiation through flow cytometry, and B is a graph quantifying it. # indicates that the GATA3+ CD25+ T cell ratio of the Th2 treatment group significantly increased compared to the IL-2 treatment group, p<0.05, and * indicates that the GATA3+ CD25+ T cell ratio of the goldenrod and Nabokja extracts mixed and extracted at a weight ratio of 4:6 of the present invention significantly decreased compared to the Th2 treatment group, p<0.05.

[0022] Figure 5 shows the results of confirming the atopic dermatitis improvement effect when the goldenrod and Nabokja extracts (200 mg / kg, 400 mg / kg) mixed and extracted at a weight ratio of 4:6 of the present invention were administered to an animal model induced with atopic dermatitis. Sham is the normal group, dermatitis is the atopic dermatitis-induced group by DNCB (2,4-Dinitrochlorobenzene) administration, and Dex is the positive control group treated with dexamethasone.

[0023] Figure 6 shows the results of confirming the effect of reducing epidermal thickness when the extracts of Scutellaria Baicalensis and Nabokja mixed and extracted at a weight ratio of 4:6 of the present invention were administered to an animal model induced with atopic dermatitis. A is a graph showing the measured epidermal thickness, and B is a photograph taken with an optical microscope after staining the epidermal layer with H&E (Hematoxylin & eosin) solution. Sham is a normal group, and an atopic dermatitis-induced group by administration of DNCB (2,4-Dinitrochlorobenzene). HN_LD is a group treated with a low concentration (200 mg / kg) of the extracts of Scutellaria Baicalensis mixed at a weight ratio of 4:6, and HN_HD is a group treated with a high concentration (400 mg / kg) of the extracts of Scutellaria Baicalensis mixed and extracted at a weight ratio of 4:6. # indicates that the epidermal thickness of the atopic dermatitis-induced group (dermatitis) significantly increased compared to the normal group (sham), p<0.05, and * indicates that the epidermal thickness of the low- and high-concentration treatment groups of the golden and Nabokja extracts mixed and extracted at a weight ratio of 4:6 of the present invention; or the positive control group (Dex); significantly decreased compared to the atopic dermatitis-induced group, p<0.05.

[0024] Figure 7 shows the results of measuring changes in (A) body weight and (B) spleen index when the golden and Nabokja extracts mixed and extracted at a weight ratio of 4:6 of the present invention were administered to an animal model in which atopic dermatitis was induced.

[0025] Figure 8 shows the results of measuring the changes in (A) transepidermal water loss (TEWL) and (B) skin hydration when the 4:6 weight ratio mixed extract of Scutellaria Baicalensis and Nabokja extract of the present invention was administered to an animal model induced with atopic dermatitis. # indicates that the transepidermal water loss of the atopic dermatitis-induced group (dermatitis) significantly increased and the skin hydration significantly decreased compared to the normal group (sham), p<0.05, and * indicates that the high-concentration treatment group of the 4:6 weight ratio mixed extract of Scutellaria Baicalensis and Nabokja extract of the present invention or the positive control group (Dex) significantly decreased and the skin hydration significantly increased compared to the atopic dermatitis-induced group, p<0.05.

[0026] Figure 9 shows the results of measuring (A) total IgE and (B) IL-4 production in serum when the 4:6 weight ratio mixed extract of Scutellaria Baicalensis and Nabokja extract of the present invention was administered to an animal model induced with atopic dermatitis. # indicates that the total IgE and IL-4 production of the atopic dermatitis-induced group (dermatitis) was significantly increased compared to the normal group (sham), p<0.05, and * indicates that the total IgE and IL-4 production of the high-concentration treatment group of the 4:6 weight ratio mixed extract of Scutellaria Baicalensis and Nabokja extract of the present invention or the positive control group (Dex) was significantly decreased compared to the atopic dermatitis-induced group, p<0.05.

[0027] In order to achieve the purpose of the present invention, the present invention provides a health functional food composition for preventing or improving atopic dermatitis, which contains extracts of gold and Nabokja as active ingredients.

[0028] The above-mentioned golden and rhododendron extracts can be prepared by a method including, but not limited to, the following steps:

[0029] (1) For gold and jasmine, a step of extracting by adding an extraction solvent;

[0030] (2) a step of filtering the extract obtained in the above step (1); and

[0031] (3) A step of concentrating the filtered extract of step (2).

[0032] In the above step (1), the extraction solvent is preferably selected from water, a lower alcohol of C1 to C4, or a mixture thereof, more preferably water, but is not limited thereto. In the above manufacturing method, the extraction method may utilize any conventional method known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction. It is preferable to extract the extraction solvent by adding 1 to 20 times the weight of the gold and the rhizome, and more preferably by adding 5 to 15 times the weight of the gold and the rhizome.

[0033] In the above method, the concentration in step (3) is preferably performed using a vacuum rotary concentrator or a vacuum rotary evaporator, but is not limited thereto.

[0034] In the above step (1), it is preferable to mix gold and jasmine in a weight ratio of 2-4:8-6, but this is not limited thereto. When mixed in the weight ratio of 2-4:8-6, the effect of improving atopic dermatitis is superior compared to when used alone or mixed in other weight ratios.

[0035] 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.

[0036] When the health functional food composition of the present invention is used as a food additive, the extracts of Scutellaria Baicalensis and Nabokja can be added as they are or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention or improvement). 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, in the case of long-term intake for the purpose of health and hygiene or health control, the amount can be below the above range, and since there is no problem in terms of safety, the active ingredient can be used in an amount above the above range. There is no particular limitation on the type of the food. Examples of foods to which the above-mentioned golden and nutmeg extracts can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the conventional sense.

[0037] 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 cyclotensin, 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. 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 within the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

[0038] In addition, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis, which contains extracts of golden and Nabokja as active ingredients.

[0039] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is one commonly used in formulation, and includes, but is not limited to, saline solution, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In addition to the above components, it may further include antioxidants, buffers, bacteriostatic agents, diluents, surfactants, binders, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, or preservatives. The pharmaceutical composition of the present invention may be administered orally or parenterally, and parenterally may be administered by injection or by application to the skin. The appropriate dosage of the pharmaceutical composition of the present invention may be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. The composition may be prepared in any one dosage form selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols.

[0040] In addition, the present invention provides an over-the-counter drug for preventing or improving atopic dermatitis, which contains extracts of gold and Nabokja as active ingredients.

[0041] The above-mentioned over-the-counter drugs may include external skin preparations and personal hygiene products. Examples include, but are not limited to, disinfectants, shower foams, mouthwashes, wet wipes, detergent soaps, hand washes, or ointments.

[0042] When the above-described pharmaceutical composition according to the present invention is used as an additive for an over-the-counter drug, the composition may be added as is or used together with other over-the-counter drugs or over-the-counter drug ingredients, and may be used appropriately according to conventional methods. The mixing amount of the active ingredient may be appropriately determined depending on the intended use.

[0043] The quasi-drug composition of the present invention may be, for example, manufactured in the form of a general emulsified formulation or a solubilized formulation. For example, it may have a formulation such as a lotion, cream, ointment, spray, oil gel, gel, oil, or aerosol. However, any formulation that exhibits the preventive or ameliorating effect of the atopic dermatitis of the present invention may be used without limitation.

[0044] In addition, the above-mentioned pharmaceutical composition may be used by appropriately mixing, as needed, oil, water, surfactant, moisturizer, lower alcohol having 1 to 4 carbon atoms, thickener, chelating agent, pigment, preservative or fragrance, etc., which are generally mixed in the pharmaceutical composition for each formulation.

[0045] In addition, the present invention provides a feed additive for preventing or improving atopic dermatitis, which contains extracts of gold and Nabokja as active ingredients.

[0046] The feed additive of the present invention may additionally include at least one selected from organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, and malic acid; phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphate (polyphosphate); and natural antioxidants such as polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid.

[0047] Animal feed additives containing the golden and Nabokja extracts of the present invention and feeds containing the same can be used together with substances such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal enzymes, digestion and absorption enhancers, growth promoters, disease preventive agents, etc. as auxiliary ingredients.

[0048] The feed additives may be administered to animals alone or in combination with other feed additives in an edible carrier. Furthermore, the feed additives may be readily administered as a top dressing, by mixing them directly into animal feed, or separately from the feed, in a separate oral formulation, by injection or transdermally, or in combination with other ingredients. Typically, a single daily dose or divided daily doses may be used, as is well known in the art. When the feed additives are administered separately from animal feed, the dosage form of the extract may be prepared as an immediate-release or sustained-release formulation by combining them with a non-toxic, pharmaceutically acceptable edible carrier, as is well known in the art. Such edible carriers may be solid or liquid, such as corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. When a solid carrier is used, the dosage form of the extract may be a tablet, capsule, powder, torokit, saccharide tablet, or a top dressing in a microdispersible form. When a liquid carrier is used, the dosage form may be a soft gelatin capsule, or a syrup, liquid suspension, emulsion, or solution. The dosage form may also contain adjuvants, such as preservatives, stabilizers, wetting or emulsifying agents, and solution promoters. Furthermore, the animal feed containing the Scutellaria Baicalensis and Nabokja extracts as feed additives may be any protein-containing organic grain meal commonly used to meet the dietary needs of animals. Such protein-containing grain meal typically consists primarily of corn, soybean meal, or a corn / soybean meal mix.

[0049] In addition, the present invention provides a veterinary composition for preventing or treating atopic dermatitis, comprising extracts of gold and Nabokja as active ingredients.

[0050] The veterinary composition comprising the extracts of Scutellaria Baicalensis and Nabokja 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 comprising the extracts of Scutellaria Baicalensis and Nabokja 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, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil.

[0051] The veterinary composition comprising the extracts of gold and rhizome of the present invention may further comprise fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc. The veterinary composition according to the present invention may be formulated using methods 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 external preparations, etc.

[0052] The veterinary composition according to the present invention may vary depending on the age, sex, and weight of the animal, but may be administered once or several times a day in an amount of 0.1 to 100 mg / kg, and the dosage may be increased or decreased depending on the route of administration, severity of the disease, sex, body weight, age, etc. Therefore, the above dosage does not limit the scope of the present invention in any way.

[0053]

[0054] 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.

[0055]

[0056] Example 1. Preparation of Golden and Nabokja Extracts

[0057] Medicinal herbs were purchased from Gwangmyeongdang, and each sample was stored in a herbal medicine refrigerator that maintained a temperature of 5.5±0.3℃ and a humidity of 55±5%. To prepare extracts of Scutellaria Baicalensis and Nabokja, the weight ratios of Scutellaria Baicalensis and Nabokja were mixed to be 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, and 9:1, and then water corresponding to 10 times the total weight was added, and extraction was performed twice under reflux cooling for 3 hours each, and the solvent was removed from the obtained filtrate using a reduced pressure concentrator using a filter paper to prepare extracts.

[0058]

[0059] Example 2. Cell viability test for extracts of goldenrod and rhizome of Nabokja

[0060] RAW 264.7 cells, an immune cell line, were purchased from the American Type Culture Collection (ATCC). RAW 264.7 cells were cultured in DMEM (Dulbecco's Modified Eagle'Media; Gibco) containing 10% FBS (Fetal Bovine Serum; Gibco) and 1% penicillin-streptomycin at 37°C and 5% CO2.

[0061] Cultured RAW 264.7 cells were treated with extracts of Scutellaria baicalensis and Nabokja prepared according to the weight ratio of Scutellaria baicalensis to Nabokja at a concentration of 100 μg / mL, and cell viability was measured using a microplate reader (Molecular Devices, Sunnyvale, CA, USA) at 490 nm using the MTS assay (CellTiter Aqueous One Solution Cell proliferation assay kit, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophynyl)-2H-tetrazolium, inner salt; MTS, Promega Co. Madison, WI, USA) technique.

[0062] As a result, as disclosed in Fig. 1, a significant decrease in survival rate was confirmed in some ratios compared to the untreated control group (Con), and the extract obtained by mixing goldenrod and Nabokja in a weight ratio of 4-1:6-9 showed a cell survival rate of more than 85%, confirming the absence of cytotoxicity.

[0063]

[0064] Example 3. Confirmation of the anti-inflammatory efficacy of golden and rhizome extracts.

[0065] 5×10 RAW 264.7 cells were seeded in a 96-well culture plate. 3 After dividing into cells / well and stabilizing for 24 hours, LPS (1 μg / ml), M2 10 5 ~10 7 CFU / ㎖, KL1 10 5 ~10 7 Each cell was treated under the condition of CFU / ㎖. After 24 hours of culture, each cell culture was separated and the change in NO (Nitric oxide) production was analyzed using the Griess Reagent System (Promega).

[0066] As a result, as disclosed in Fig. 2, NO production was significantly reduced in the extracts obtained by mixing the extracts of golden and Nabokja in weight ratios of 4:6, 3:7, and 2:8 compared to the LPS-only treatment group, and in particular, the highest level of reduction was confirmed at the weight ratio of 4:6.

[0067]

[0068] Example 4. Comparison of cell viability and anti-inflammatory efficacy of golden extract and Nabokja extract alone and golden extract and Nabokja extract combined

[0069] The cell viability and anti-inflammatory efficacy of the golden and Nabokja extracts, which were extracted by mixing them in a weight ratio of 4:6, which showed the best anti-inflammatory efficacy, and the golden and Nabokja extracts alone were measured and compared. The experimental method was as described above.

[0070] As a result, as shown in Fig. 3A, NO production was significantly reduced in the group treated with only the gold extract (Hwang 40); and the group treated with the gold and Nabokja extracts mixed and extracted at a weight ratio of 4:6 (Hwang 40 / Na 60); compared to the group treated with only the LPS, and as shown in Fig. 3B, in the group treated with only the gold extract, cell viability was reduced by about 50% or more, indicating cytotoxicity.

[0071] Meanwhile, in the group treated with Nabokja extract alone, there was no statistically significant reduction in the NO production inhibition effect compared to the LPS alone treatment group (Fig. 3A).

[0072] Therefore, it was confirmed that the golden and Nabokja extracts extracted by mixing them in a weight ratio of 4:6 of the present invention were non-cytotoxic and had excellent anti-inflammatory effects.

[0073]

[0074] Example 5. Confirmation of the immunomodulatory effects of golden and rhizome extracts.

[0075] To verify the hypersensitivity immune response suppression effect of the extracts of Scutellaria Baicalensis and Nabokja mixed at a weight ratio of 4:6, which have the best anti-inflammatory efficacy, splenic CD4+ T cells were separated from the spleen of mice using a magnetic bead cell sorting method, and the purity of the cells was checked before use in the experiment. To induce differentiation of the separated cells, they were cultured in a medium containing beads coated with anti-CD3 / CD28 and IL2, IL4, and anti-INF-γ antibodies, and the degree of cell differentiation and activation according to the presence or absence of the test substance was confirmed through flow cytometry.

[0076] As a result, as disclosed in Fig. 4, when CD4+ T cells were induced to differentiate into type 2 helper T cells through cytokine and antibody treatment, and the extracts of goldenrod and Nabokja mixed and extracted at a weight ratio of 4:6 of the present invention were treated, it was confirmed that the activity and differentiation of acquired immune cells were suppressed in a concentration-dependent manner.

[0077]

[0078] Example 6. Confirmation of the anti-atopic efficacy of extracts of Scutellaria baicalensis and Nabokja in an animal model of atopic dermatitis.

[0079] To minimize the effects of micro-wounds caused by hair removal, a skin inflammation experiment was conducted using nude mice, and atopic dermatitis was induced by applying DNCB (2,4-Dinitrochlorobenzene) solution to the back. The experimental groups were divided into a total of five groups: a normal group (Sham), an atopic dermatitis-induced group (dermatitis), a low-concentration treatment group of Scutellaria baicalensis and Nabokja extracts (HN_LD, 200 mg / kg), a high-concentration treatment group of Scutellaria baicalensis and Nabokja extracts (HN_HD, 400 mg / kg), and a positive control group (Dex, dexamethasone). The test substances and control drugs were administered orally using mouse zones.

[0080] At the end of the test, the skin condition was visually observed, and transepidermal water loss (TEWL) and skin hydration were measured using a skin analyzer. The serum portion of the blood obtained from each experimental group was separated and the levels of inflammatory mediators contained in the blood were measured, and the secretion levels of immunoglobulin E (IgE) and interleukin 4, which are reported to be highly related to hypersensitivity skin inflammation reactions, were measured using ELISA. In addition, the skin tissue obtained from each experimental group was fixed in a 10% formalin solution, paraffin blocks were made, and tissue sections were made. After H&E staining, histopathological analysis was performed.

[0081] As a result, as disclosed in Fig. 5, it was confirmed that the animal model in which atopic dermatitis was induced by DNCB application showed improvement in atopic dermatitis symptoms (skin formation, dryness, swelling, etc.) due to administration of the extracts of Scutellaria baicalensis and Nabokja of the present invention, and when the skin thickness of the area where atopic dermatitis was induced was measured, it was confirmed that the epidermal layer thickened by DNCB application was significantly reduced by administration of the extracts of Scutellaria baicalensis and Nabokja of the present invention (Fig. 6).

[0082] In addition, it was confirmed that the increased spleen index due to the induction of atopic dermatitis was reduced by the administration of the extracts of Golden and Nabokja of the present invention (Fig. 7), and that the increased transepidermal water loss and decreased skin moisture due to the induction of atopic dermatitis were recovered by the administration of the extracts of Golden and Nabokja of the present invention (Fig. 8).

[0083] Lastly, as a result of analyzing the secretion amount of total IgE and IL-4, which are indicators related to atopic dermatitis, it was confirmed that the increased content of total IgE and IL-4 due to induction of atopic dermatitis was reduced by administration of the extracts of golden and Nabokja of the present invention (Fig. 9).

Claims

1. A health functional food composition for preventing or improving atopic dermatitis, comprising extracts of gold and Nabokja as active ingredients.

2. A health functional food composition for preventing or improving atopic dermatitis, characterized in that the extraction solvent of the extract in paragraph 1 is water, a lower alcohol of C1 to C4, or a mixture thereof.

3. A health functional food composition for preventing or improving atopic dermatitis, characterized in that the extract of gold and Nabokja in the first paragraph is a mixture of gold and Nabokja extracted in a weight ratio of 2 to 4:8 to 6.

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

5. A pharmaceutical composition for preventing or treating atopic dermatitis, comprising extracts of gold and Nabokja as active ingredients.

6. A pharmaceutical composition for preventing or treating atopic dermatitis, characterized in that, in addition to the effective ingredient in paragraph 4, it further comprises a pharmaceutically acceptable carrier, excipient or diluent.

7. A pharmaceutical composition for preventing or treating atopic dermatitis, characterized in that the composition in claim 4 is prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols.

8. A non-prescription drug for preventing or improving atopic dermatitis containing extracts of gold and Nabokja as active ingredients.

9. Feed additive for preventing or improving atopic dermatitis containing extracts of goldenrod and Nabokja as active ingredients.

10. A veterinary composition for the prevention or treatment of atopic dermatitis containing extracts of goldenrod and Nabokja as active ingredients.

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