KR20220098594A - Composition for preventing, improving or treating atopic or allergic dermatitis comprising deer antler fermentation products as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating atopic or allergic dermatitis comprising a velvet antler fermentation product as an active ingredient. The velvet antler fermentation product according to the present invention does not show cytotoxicity in weakened cells induced by inflammation while inhibiting atopic dermatitis or allergic dermatitis induced by the expression of MDC or TARC, and thus, has excellent stability and can be used as a composition for preventing, alleviating or treating the atopic dermatitis or allergic dermatitis. In addition, since the present invention is a composition derived from natural products, the composition of the present invention can be safely used without side effects, and can be usefully used for medicines or foods.

Description

A composition for preventing, improving or treating atopic or allergic dermatitis comprising deer antler fermentation products as an active ingredient, comprising fermented antler as an active ingredient

The present invention relates to a composition for preventing, improving or treating atopic dermatitis or allergic dermatitis, comprising a fermented antler as an active ingredient.

With the deepening of industrialization and urbanization, changes in diet and residential life, and exposure to chemical and biological harmful substances caused by environmental pollution have led to a significant increase in allergic diseases in various age groups. The human body shows a sensitive reaction to a harmless external environment, and it appears in respiratory diseases such as asthma or rhinitis, or skin diseases such as contact dermatitis and atopic diseases.

In particular, the skin is in direct contact with external allergens by air, food, etc., causing a sensitive immune response. Allergic dermatitis is an inflammatory disease caused by continuous exposure to external antigens such as food, bacteria, mites, and climate environmental factors. In order to improve and treat these diseases, there is a method of applying an appropriate active ingredient to the affected area of dermatitis or administering it to the whole body while avoiding exposure and contact with allergens and keeping the skin clean.

However, synthetic allergic dermatitis inhibitors are expensive, and both pharmaceutical and cosmetic methods cannot prevent the recurrence of lesions. There is an urgent need to develop therapeutic agents using natural materials for this purpose.

In addition, atopic dermatitis is a chronic inflammatory disease that occurs before the development of asthma and allergic diseases, and the symptoms differ depending on the concentration, quantitative change or activity level of chemokines, and Th2 type lymphocytes are found in the lesion area. It is also referred to as the phenomenon of invasion through the lesionalskin. Atopic dermatitis is a phenomenon in which the proliferation of keratinocytes is abnormally caused by immunological factors. When keratinocytes are exposed to IFN-γ (interferon-γ) or TNF-α (tumor necrosis factor-α), TARC , it can be confirmed that chemokines such as MDC and CTACK are abnormally expressed. This phenomenon is because an inflammatory reaction occurs and the penetration of monocytes/T cells into the skin increases.

Although the exact pathophysiology of atopic dermatitis is not yet fully understood, it is believed that immunological and non-immunological mechanisms are involved along with genetic predisposition. Exogenous atopic dermatitis, which accounts for most of atopic dermatitis, is caused by an immune mechanism related to IgE, and there are many reports that a delayed immune response caused by T cell abnormality is involved rather than an immediate immune response to a specific allergen. In addition, these days, Th2-related cytokines such as IL-4, which induce the production of IgE from B cells, are reported to be the cause of atopy (JS Kang et al , Ihhibition of atopic dermatitis by topical application of silymarin in NC). /Nga mice Int Immunopharm (2008) 8:1475-1480).

Since it is not easy to find and treat such atopic dermatitis, it is common to avoid triggering factors and control the symptoms of atopic dermatitis through appropriate treatment, rather than aiming for a cure. For atopic dermatitis, drug therapy, such as steroids, antihistamines, and antibiotics, is mainly used. Steroids (corticosteroids) have anti-inflammatory and immunosuppressive effects and have excellent effects, but steroids may cause adverse effects such as growth inhibition or side effects, urea peroxide, etc. may cause excessive skin irritation, and antibiotics such as antihistamines may cause side effects such as resistance to bacteria and photosensitivity. There is a high problem. In addition, when applied to the skin for a long period of time, there is a concern that serious side effects such as telangiectasia and/or increase or expansion of the stratum corneum may occur.

Recently, the focus has been on the search for natural substances that have an effect on atopy. Representatively, mugwort extract (Korean Patent Registration No. 10-0377262), Reishi mushroom, Eugeunpi, licorice, Baekbongryeong, white sage and baeknyeoncho extract (Korea Patent Registration No. 10-0517465), Evening primrose oil, Aloe, Wood vinegar, Violet, Extracts of jujube, matsutake mushroom, elm, ginseng, green tea, duchung, bokbunja, omija, mugwort, pogongyeong, trifolieae, astragalus, hagograss, sea pine, gold and hyssop (Korea Patent Registration No. 10-0451444), celadon leaf and red jasmine Leaflet extract (Korean Patent Registration No. 10-0454752), Geonryul, Uiin, Omija, Gilkyung, Nabokja, Maekmundong and Seokchangpo extracts (Korean Patent Registration No. 10-0483539), Lavender, Eucalyptus and Tea Tree Oil (Korean Patent) Registration No. 10-0597997), etc.), and in addition to that, it is known that the above-mentioned saengsaeng, Hapwanpi, Changjae, malt, etc. are effective.

However, the natural substances have a disadvantage in that their efficacy is insignificant, or although they are effective, they show allergic hypersensitivity reactions in sensitive skin, limiting their use.

On the other hand, antler ( Cornu cervi pantotrichum , antler ) refers to the cartilage tissue that is regenerated every year with young horns that are dense and not ossified with the hairs of plum, maroc, and closely related animals belonging to the deer family. It is called antler, and in Eastern countries such as China and Japan, including Korea, antler has been widely used as the best blood tonic for a long time.

As such, various methods have been studied for ingesting antlers having various effects, and among them, methods for improving absorption and efficacy by fermenting antlers have been developed.

As a result of repeated research on the fermented antler, the inventors of the present invention confirmed that the fermented antler exhibits a very excellent therapeutic effect on atopic dermatitis or allergic dermatitis, and completed the present invention.

KR 10-2011-0051162 A

It is an object of the present invention to provide a composition for preventing, improving or treating atopic dermatitis or allergic dermatitis, comprising a fermented deer antler as an active ingredient.

In order to achieve the above object, the present invention provides a food composition for preventing or improving atopic dermatitis or allergic dermatitis comprising a fermented deer antler as an active ingredient.

According to an embodiment of the present invention, the fermented antler may be obtained by fermenting the antler extract with lactic acid bacteria or Bacillus subtilis .

According to an embodiment of the present invention, the antler extract may be an extract of water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

According to an embodiment of the present invention, the lactic acid bacteria are Lactobacillus genus, Bifidobacterium genus, Leuconostok genus, Pediococcus genus, Weissella genus, Staphylococcus genus, Erococcus genus and Enterococcus genus among lactic acid bacteria. It may be any one or more selected lactic acid bacteria.

According to an embodiment of the present invention, the lactic acid bacteria may be any one or more selected from Leukonostok mecenteroides, Lactobacillus plantarum, and Lactobacillus paraplantarum.

According to an embodiment of the present invention, the composition can inhibit the expression of MDC (Marcrophage-derived chemokine) and TARC (Thymus and activation regulated chemokine).

According to one embodiment of the present invention, the composition can reduce the amount of IgG in serum.

In addition, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis or allergic dermatitis comprising a fermented antler as an active ingredient.

In addition, the present invention provides a cosmetic composition for preventing or improving atopic dermatitis or allergic dermatitis comprising fermented antler as an active ingredient.

The fermented deer antler according to the present invention suppresses atopic dermatitis or allergic dermatitis induced by the expression of MDC or TARC, but does not show cytotoxicity in weakened cells induced by inflammation, and thus has excellent stability against atopic dermatitis or allergic dermatitis. It can be used as a composition for prevention, improvement or treatment. In addition, since the present invention is a composition derived from a natural product, it can be safely used without side effects, and thus can be usefully used in medicines or foods.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for preventing, improving or treating atopic dermatitis or allergic dermatitis, comprising a fermented antler as an active ingredient.

Atopic dermatitis or allergic dermatitis is an inflammatory skin disease accompanied by itching. In keratinocytes (HaCaT), high concentrations of inflammation-inducing cytokines such as TNF-α and IFN-γ exist, and these inflammation-inducing cytokines are TARC (thymus). and activation-regulated chemokines) and increases MDC production. As a result, the concentration of cytokines was found to be closely related to the symptoms of atopic dermatitis or allergic dermatitis accompanied by itching, and TARC and MDC are reliable biomarkers that best reflect these diseases.

Accordingly, in the present invention, cells treated with TARC and MDC, which are biomarkers shown in inflammatory diseases such as atopic dermatitis or allergic dermatitis, such as TARC and MDC, and treated with inflammation-inducing substances TNF-α and/or IFN-γ. As a result of efforts to discover new substances that do not exhibit cytotoxicity in It was confirmed that it is possible.

Specifically, the present invention provides a food composition for preventing or improving atopic dermatitis or allergic dermatitis comprising a fermented deer antler as an active ingredient.

As used herein, "fermented antler" may be fermented with lactic acid bacteria or Bacillus subtilis ( Bacillus subtilis ) of the antler extract. Preferably, the deer antler is selected from any one or more lactic acid bacteria selected from the genus Lactobacillus, Bifidobacterium, Leuconostok, Pediococcus, Weissella, Staphylococcus, Erococcus and Enterococcus. , or may be fermented with Bacillus subtilis. More preferably, the antler is used as any one or more lactic acid bacteria or Bacillus subtilis selected from Leuconostoc mesenteroides , Lactobacillus plantarum and Lactobacillus paraplantarum . It may be fermented. Most preferably, antler leuconostoc mesenteroides SST-9962 [Accession No.: KCCM12712P], Lactobacillus paraplantarum SST-9961 [Accession No.: KCCM12711P] and Bacillus subtili It may be fermented with any one strain selected from Bacillus subtilis SST-9960 [Accession No.: KCCM 11402P].

The "fermented antler" may be used without limitation, fermented by a conventional fermentation method known in the art.

On the other hand, the antler extract includes not only the crude extract obtained by treating the extraction solvent on the antler, which is an extraction raw material, but also the processed product of the crude extract. For example, the deer antler extract may be prepared in a powder state by an additional process such as distillation under reduced pressure, freeze-drying or spray-drying. The extract also includes a fraction obtained by additionally fractionating the crude extract.

The antler extract is an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and the extraction method does not need to be particularly limited.

As a preferred embodiment of the present invention, when water is used as a solvent, hot water extraction is preferred. For example, it can be extracted in water of 80 to 130 ℃, preferably 90 to 110 ℃ 0.5 to 24 hours, preferably 1 to 6 hours. And, even if hot water is not used as the solvent, that is, the antler component can be extracted from the diluted solution obtained by diluting and mixing the antler powder with cold water or water at room temperature. When extracting the antler component using cold water or room temperature water, it may be extracted separately from fermentation, or may be inoculated with lactic acid bacteria or Bacillus subtilis so that the antler component is eluted from the antler during the fermentation process.

As a preferred embodiment of the present invention, an extract with an aqueous alcohol solution having 1 to 4 carbon atoms may be used. For example, extraction may be performed with an aqueous alcohol solution such as ethanol, methanol, isopropanol, etc., preferably 20 to 80% by weight of an aqueous alcohol solution, more preferably 50 to 70% by weight of an aqueous alcohol solution. In the case of using an alcohol extract, before inoculating lactic acid bacteria or Bacillus subtilis, the alcohol contained in the alcohol extract is vaporized to lower the alcohol content or the alcohol extract is concentrated and dried, and then dissolved in water for use.

Also, in the present invention, the antler extract includes a diluent obtained by diluting antler powder in water in a broad sense. In the test example of the present invention, atopic dermatitis or allergic dermatitis of antler fermented product obtained by fermenting antler hot water extract or antler ethanol aqueous solution extract with lactic acid bacteria or Bacillus subtilis, compared to antler fermented product fermented with lactic acid bacteria or Bacillus subtilis. Although the preventive, ameliorating or therapeutic effect of the antlers was somewhat low, it was confirmed that the effect was significantly increased compared to the unfermented deer antler extract.

Before inoculating the antler extract with lactic acid bacteria or Bacillus subtilis, a lactic acid bacteria nutrient source such as a protein source, a carbohydrate source, a vitamin or a mineral may be additionally mixed in order to promote the growth of the strain. The strain nutrient source may use a commercially available medium or may be individually added only the necessary nutrient source.

In addition, the antler extract, or the mixture of the antler extract and the strain nutrient source may be heat-sterilized before inoculation with lactic acid bacteria or Bacillus subtilis.

The solid content of the deer antler extract does not need to be particularly limited, but is 1 to 15% by weight if there is no other concentration process after extraction, and it may be used directly, concentrated, or diluted after concentration or drying.

In the present invention, the lactic acid bacteria is any one or more selected from Lactobacillus, Bifidobacterium, Leuconostok, Pediococcus, Weissella, Staphylococcus, Erococcus and Enterococcus lactic acid bacteria. It may be lactic acid bacteria. Preferably, the lactic acid bacterium may be any one or more selected from Leukonostok mecenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum, Lactobacillus saccharis and Pediococcus pentosaceus, more preferably the The lactic acid bacteria may be any one or more selected from Leukonostok mecenteroides, Lactobacillus plantarum, and Lactobacillus paraplantarum, and more preferably, the lactic acid bacteria is Leukonostok mecenteroides and Lactobacillus paraplantarum. It may be any one or more selected.

The strain may be added as a strain itself or as a culture solution. The culture solution may include the culture solution itself in which the strain is cultured in a liquid medium, and a concentrate obtained by concentrating the culture solution.

In a preferred embodiment of the present invention, the fermentation strain, particularly the lactic acid bacteria or Bacillus subtilis strain, is 1×10 ⁶ to 1×10 ⁹ cfu/mL, preferably 1×10 ⁶ to 1×10 ⁸ cfu/mL, More preferably, 0.3 to 3.0 parts by weight, preferably 1.0 to 2.5 parts by weight, more preferably 1.5 parts by weight of the culture solution contained at a concentration of 1×10 ⁷ to 1×10 ⁸ cfu/mL based on 100 parts by weight of the antler extract. to 2.5 parts by weight may be added.

In the present invention, the "fermented antler" for producing the desired effect significantly, (1) put the antler extract in a fermentation tray, and 0.3 to 3.0 weight of the lactic acid bacteria or Bacillus subtilis culture based on 100 parts by weight of the antler extract After adding (2) at 20 to 40°C for 18 to 96 hours, preferably 18 to 96 hours, more preferably 18 to 72 hours, still more preferably 18 to 48 hours, still more preferably 18 to 36 hours. Time, more preferably, it may be prepared by fermentation for 20 to 30 hours.

In particular, (3) after the fermentation at 2 to 10 ° C. for 5 to 24 hours after low-temperature aging, (4) 25 to 35 ° C., preferably 28 to 33 ° C. for 4 to 8 hours air drying step It is more preferable in terms of effect to carry out additionally. According to an embodiment of the present invention, it was specifically confirmed that the improvement effect of atopic dermatitis or allergic dermatitis is further enhanced when the fermented antler fermented product is aged at a low temperature.

In addition, the "fermented antler" prepared by the above method has a higher nutritional content than the fermented antler produced by another method, or the fermented deer antler produced and fermented by a strain other than lactic acid bacteria or Bacillus subtilis. It was specifically confirmed that the improvement effect of atopic dermatitis or allergic dermatitis was further enhanced.

In addition, since the lactic acid bacteria among the strains for the fermentation have excellent growth in the deer antler extract, there is an advantage in that the fermentation time is short compared to other strains including the Bacillus subtilis strain.

Meanwhile, the fermented deer antler may be a fermented product containing microorganisms, or may be a fermented product from which the cells are removed. The fermented product from which the cells are removed may be sterilized to contain dead cells, or may be a filtrate or centrifugation supernatant obtained by removing the cells through filtration or centrifugation. The prevention, improvement or treatment effect of atopic dermatitis or allergic dermatitis of the present invention is to bioconvert the components contained in the deer antler extract into easy-to-use components in the body while the microorganisms proliferate, or to prevent atopic dermatitis or allergic dermatitis , it is expected because a new active ingredient having an improving or therapeutic effect is produced, and it is significantly higher than the effect that can be obtained by adding a simple strain itself. The fermented antler may be used as a liquid fermented product by itself, or may be dried and powdered for use.

In the present specification, "comprising as an active ingredient" means including an amount sufficient to achieve prevention, improvement, or therapeutic efficacy or activity of atopic dermatitis or allergic dermatitis of the fermented product of the present invention.

As used herein, the term “prevention” refers to anything that inhibits or delays the atopic dermatitis or allergic dermatitis.

As used herein, the term “improvement” means at least reducing a parameter related to a condition to be treated, for example, the degree of symptoms by administration of a composition comprising the fermented antler of the present invention.

As used herein, the term “treatment” means reversing, alleviating, inhibiting the progression, or preventing the symptoms of atopic dermatitis or allergic dermatitis, unless otherwise stated.

As used herein, the term "atopic dermatitis" is a recurrent chronic skin disease accompanied by severe itching that mainly begins in infancy or childhood, and is a very common skin disease accompanied by a family history. Symptoms begin in infancy and infancy, especially around 2 months of age, and about 50% of them occur before 2 years of age, and most of them appear before 5 years of age. . In some patients, symptoms relieve or heal spontaneously with growth, and more than half of patients who develop in infancy improve before 2 years of age. The shape and distribution of skin lesions is characteristic, and it is accompanied by pruritus (itching), dry skin, and characteristic eczema. In infancy, it begins as eczema on the face and on the outstretched side of the limbs, but as you grow older, it characteristically appears in the form of eczema on the bent part of the arm and the bent part behind the knee. In adults, lichenification occurs when the skin at the fold is thickened, and eczema occurs on the face, chest, and nape of the neck other than the extremities compared to infancy and childhood.

As used herein, the term “allergic dermatitis” is a recurrent eczema lesion accompanied by a chronically progressive intense itching. The rash tends to appear on curved areas such as the face, neck, elbows and knees, and when the aggravation is severe, it may spread to the whole body. Specific symptoms or diseases include erythema, papules, lichenification, dryness, itchy skin (pruritus), pruritic urticaria, cheilitis, keratosis pilaris, pityriasis pityriasis white, and white skin feat.

The composition of the present invention does not exhibit cytotoxicity in cells such as HaCaT induced by inflammatory cytokines TNF-α and/or IFN-γ by including the fermented deer antler as an active ingredient, and TNF-α and/or IFN -γ suppresses and inhibits the expression of biomarkers of TARC and MDC in cells induced by atopic dermatitis or allergic dermatitis, thereby having a remarkable effect in preventing or improving atopic dermatitis or allergic dermatitis. The atopic dermatitis or allergic dermatitis may be induced by MDC or TARC expression.

In the following test examples, it was specifically confirmed that the fermented deer antler of the present invention can exhibit significant improvement or therapeutic effects for both atopic dermatitis or allergic dermatitis.

The food composition for the prevention or improvement of atopic dermatitis or allergic dermatitis of the present invention includes all forms of nutritional supplements, health functional foods, food additives and feeds, and human Alternatively, animals, including livestock, are intended to be eaten.

The food composition may be formulated in the form of capsules, tablets, powders, granules, liquids, pills, flakes, pastes, syrups, gels, jellies or bars of the fermented antler of the present invention. The food composition has the advantage of not having side effects that may occur during long-term administration, unlike general drugs.

In addition, the food composition can be prepared in various forms according to conventional methods known in the art. Common foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat and their processed foods (eg, ham, sausages) Corn beef, etc.), breads and noodles (eg udon noodles, soba noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (eg butter, cheese, etc.), edible vegetable oils and fats, margarine , vegetable protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.) can be prepared by adding the fermented deer antler. In addition, the nutritional supplement is not limited thereto, but may be prepared by adding the fermented antler to capsules, tablets, pills, and the like. In addition, the health functional food is not limited thereto, but for example, the fermented deer antler itself can be prepared in the form of tea, juice, and drink and liquefied for drinking (health drink), granulated, encapsulated and powder It can be eaten raw. In addition, in order to use the fermented antler fermented product in the form of a food additive, it may be prepared as a powder or extracted and then used in the form of a concentrate.

When the composition for preventing or improving atopic dermatitis or allergic dermatitis of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients, like a conventional drink. . The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as taumatine, stevia extract; A synthetic sweetener such as saccharin or aspartame may 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 mL of the composition of the present invention.

The fermented deer antler of the present invention may be contained as an active ingredient in the food composition, and the amount may be an amount effective to achieve a preventive, ameliorating or therapeutic effect for atopic dermatitis or allergic dermatitis. Specifically, the fermented deer antler is 5 to 250 μg/ml, preferably 10 to 100 μg/ml, more preferably 50 to 100 μg/ml, most preferably 100 μg/ml with respect to the total content of the cosmetic composition. may be included in the concentration of In this case, if the content of the fermented antler is less than the lower limit, the cell viability is excellent, but the improvement effect of atopic dermatitis or allergic dermatitis may not appear to a desired degree. Conversely, when the upper limit is exceeded, the improvement effect of atopic dermatitis or allergic dermatitis does not increase as the concentration increases, or it may be toxic. On the other hand, as a result of the in vitro experiment, when the concentration of the fermented antler of the present invention was within the above range, there was no side effect such as cytotoxicity while showing a significant effect on the improvement of atopic dermatitis or allergic dermatitis.

The food composition of the present invention can be prepared by mixing the antler fermented product with other active ingredients known to be effective in preventing, improving or treating atopic dermatitis or allergic dermatitis.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, It may contain glycerin, alcohol or a carbonation agent and the like. In addition, the health food of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis or allergic dermatitis comprising a fermented antler as an active ingredient. The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant other than the fermented antler, and the adjuvant includes an excipient, a disintegrant, a sweetener, a binder, a coating agent, a swelling agent, a lubricant, A lubricant or flavoring agent may be used. As used herein, the terms “fermented antler” and “atopic dermatitis or allergic dermatitis” are the same as described above.

In addition, the pharmaceutical composition for the prevention or treatment of atopic dermatitis or allergic dermatitis of the present invention may be formulated including one or more pharmaceutically acceptable carriers.

The pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration.

Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like.

Carriers for parenteral administration may also include water, suitable oils, saline, aqueous glucose and glycols, and the like. In addition, it may further include a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. As other pharmaceutically acceptable carriers, reference may be made to those described in the following literature (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The pharmaceutical composition of the present invention may be administered to mammals including humans by any method. For example, it may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration.

The pharmaceutical composition of the present invention may be formulated as a formulation for oral administration or parenteral administration according to the administration route as described above. When formulated, one or more buffers (eg, saline or PBS), antioxidants, bacteriostatic agents, chelating agents (eg, EDTA or glutathione), fillers, bulking agents, binders, adjuvants (eg, aluminum hydroxide) side), suspending agents, thickening agents, wetting agents, disintegrating agents or surfactants, diluents or excipients.

Formulations for oral administration include tablets, pills, powders, granules, solutions, gels, syrups, slurries, suspensions or capsules, and such solid preparations include at least one excipient, for example, in the pharmaceutical composition of the present invention. , Starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose , methyl cellulose, sodium carboxymethyl cellulose, and hydroxypropylmethyl-cellulose or gelatin may be mixed and prepared. For example, tablets or dragees can be obtained by combining the active ingredient with solid excipients, grinding them, adding suitable auxiliaries, and processing them into a mixture of granules.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions or syrups, and various excipients, such as wetting agents, sweetening agents, fragrances, or preservatives, in addition to water or liquid paraffin, which are commonly used simple diluents, may be included. .

In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, and an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifier and a preservative may be further included. .

When administered parenterally, the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations and nasal inhalants together with suitable parenteral carriers. In the case of the injection, it must be sterilized and protected from contamination of microorganisms such as bacteria and fungi. For injection, examples of suitable carriers include, but are not limited to, water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and/or a solvent or dispersion medium containing vegetable oil. can More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with triethanolamine or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. etc. can be used. In order to protect the injection from microbial contamination, it may further include various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, in most cases, the injection may further contain an isotonic agent such as sugar or sodium chloride.

In the case of transdermal administration, forms such as ointment, cream, lotion, gel, external solution, pasta, liniment, and air are included. As used herein, "transdermal administration" means that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin by topically administering the pharmaceutical composition to the skin.

In the case of administration by inhalation, using a propellant suitable for the composition according to the invention, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas, it can be dispensed from a pressurized pack or nebulizer. It can be conveniently delivered in the form of an aerosol spray. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges used in inhalers or insufflators may be formulated to contain a powder mixture of a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a recipe commonly known to all pharmaceutical chemistry.

When the pharmaceutical composition of the present invention contains the fermented antler in an effective amount, it is possible to provide a desirable preventive, ameliorating or therapeutic effect of atopic dermatitis or allergic dermatitis. In the present specification, the term 'effective amount' refers to an amount that exhibits a higher reaction than that of a negative control group, and preferably refers to an amount sufficient to improve or treat atopic dermatitis or allergic dermatitis. The fermented deer antler is at a concentration of 5 to 250 μg/ml, preferably 10 to 100 μg/ml, more preferably 50 to 100 μg/ml, and most preferably 100 μg/ml with respect to the total content of the pharmaceutical composition. may be included. In this case, if the content of the fermented antler is less than the lower limit, the cell viability is excellent, but the improvement effect of atopic dermatitis or allergic dermatitis may not appear to a desired degree. Conversely, when the upper limit is exceeded, the improvement effect of atopic dermatitis or allergic dermatitis does not increase as the concentration increases, or it may be toxic. On the other hand, as a result of the in vitro experiment, when the concentration of the fermented antler of the present invention was within the above range, there was no side effect such as cytotoxicity while showing a significant effect on the improvement of atopic dermatitis or allergic dermatitis. The effective amount of the fermented deer antler contained in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.

The total effective amount of the pharmaceutical composition of the present invention may be administered to a patient as a single dose, and may be administered by a fractionated treatment protocol in which multiple doses are administered for a long period of time. . The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease.

A suitable dosage of the pharmaceutical composition varies depending on factors such as formulation method, administration mode, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate, and response sensitivity of the patient, usually A skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment, the daily dose of the pharmaceutical composition is 0.001 to 10 g/kg.

The pharmaceutical composition for the prevention and treatment of atopic dermatitis or allergic dermatitis of the present invention may also be provided in the form of an external preparation containing fermented antler as an active ingredient.

When the pharmaceutical composition for the prevention and treatment of atopic dermatitis or allergic dermatitis of the present invention is used as an external preparation for skin, a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, and a stabilizer , foaming agent, fragrance, surfactant, water, ionic emulsifier, non-ionic emulsifier, filler, sequestering agent, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic It may contain adjuvants commonly used in the field of dermatology, such as active agents, lipophilic active agents, or any other ingredients commonly used in external preparations for skin, such as lipid vesicles. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.

When the pharmaceutical composition for the prevention and treatment of atopic dermatitis or allergic dermatitis of the present invention is provided as an external preparation for the skin, the present invention is not limited thereto, but may be in the form of an ointment, a patch, a gel, a cream, or a spray.

In addition, the present invention provides a cosmetic composition for preventing or improving atopic dermatitis or allergic dermatitis comprising fermented antler as an active ingredient. In the present invention, the terms "fermented green tea" and "atopic dermatitis or allergic dermatitis" are as described above.

In a preferred embodiment of the present invention, the fermented deer antler may be present in the composition in any amount desired to provide effective prevention or improvement efficacy against atopic dermatitis or allergic dermatitis. According to one embodiment, the fermented deer antler is 5 to 250 μg/ml, preferably 10 to 100 μg/ml, more preferably 50 to 100 μg/ml, most preferably 100 μg, based on the total content of the cosmetic composition. It may be included at a concentration of /ml. In this case, if the content of the fermented antler is less than the lower limit, the cell viability is excellent, but the improvement effect of atopic dermatitis or allergic dermatitis may not appear to a desired degree. Conversely, when the upper limit is exceeded, the improvement effect of atopic dermatitis or allergic dermatitis does not increase as the concentration increases, or it may be toxic. That is, since the fermented deer antler of the present invention is included in the above range in the composition, an excellent preventive or ameliorating effect can be realized for atopic dermatitis or allergic dermatitis, it has formulation and product stability, and has an excellent effect with an optimal content in terms of economy. can perform

Components included in the cosmetic composition include components commonly used in cosmetic compositions in addition to the fermented deer antler as an active ingredient, for example, conventional adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, and fragrances. may further include.

The cosmetic composition is a suspension, emulsion, paste, gel, cream, atopic soap, cleansing foam, cleansing cream, cleansing water, bath agent, skin softener, skin toner, lotion, cream, essence, astringent, emulsion, gel, lipstick, It may be in any one formulation selected from the group consisting of spray, shampoo, rinse, treatment, body cleanser, pack, massage agent, face powder, compact, foundation, two-way cake, and makeup base.

When the formulation of the cosmetic composition is a paste, cream or gel, vegetable oil, wax, starch, cellulose, silicon, bentonite, silica, zinc oxide, etc. may be further included as a carrier component, and when the formulation is a powder, lactose as a carrier component , silica, aluminum hydroxide, may further include powders such as calcium silicate. When the formulation of the cosmetic composition is an emulsion, a solvent, solubilizer or emulsifier may be further included as a carrier component, for example, water, ethanol, glycerin, ethyl carbonate, ethyl acetate, propylene glycol, butylene glycol. , 1,2-hexanediol, glycerol fatty esters, and sorbitan fatty acid esters.

When the formulation of the cosmetic composition is a suspension, as a carrier component, a liquid diluent such as water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and a suspending agent such as polyoxyethylene sorbitan ester , microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, and the like may be further included.

The cosmetic composition of the present invention may be used alone or in overlapping application, or may be used in overlapping application with other cosmetic compositions other than the present invention. In addition, the cosmetic composition according to the present invention can be used according to a conventional method of use, and the number of times of use can be changed according to the skin condition or taste of the user.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these Examples are intended to illustrate the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereby.

<Example>

Preparation Example 1: Preparation of deer antler hot water extract

After selecting and washing with water, domestic antler powder was uniformly pulverized with a grinder to prepare an antler powder with an average particle size of 300 μm, which was used while refrigerated.

The deer antler powder and 10 times the weight of distilled water were put in a reflux extractor, extracted twice at 100° C. for 3 hours, cooled and concentrated under reduced pressure to obtain a hot water extract of deer antler. The antler hot water extract had a sugar content of 1.6 brix, a reducing sugar content of 0.24%, and a protein content of 0.31%.

Preparation Example 2: Preparation of deer antler ethanol aqueous solution extract

The antler antler powder of Preparation Example 1 and 4 times the weight of 50% by weight of an aqueous ethanol solution were added, and the mixture was extracted at room temperature for 3 days, and then concentrated under reduced pressure to obtain an aqueous ethanol solution of antler extract. The sugar content of the ethanol aqueous solution extract of the deer antler was 1.7 Brix.

Preparation Example 3: Preparation of antler water diluted solution

Water was mixed with water so that the antler powder of Preparation Example 1 was 2.0% by weight to prepare a water dilution solution for antler.

Examples 1-2: Preparation of fermented deer antler

The antler extract obtained by diluting the deer antler extract of Preparation Example 1 with distilled water 4 times the weight, or the diluted solution of the deer antler water of Preparation Example 3 was sterilized at 121° C., 1.5 atmospheres, and 15 minutes. In addition, each of the lactic acid bacteria of Table 1 was inoculated into the MRS broth medium and activated by stationary culture at 37 ° C. for 24 hours to prepare a lactic acid bacteria culture solution.

Then, 1.5% by weight of glucose was added to the deer antler extract, and each of the prepared lactic acid bacteria cultures (1×10 ⁸ CFU/mL) was inoculated at 1.0% by weight, followed by stationary culture at 37° C. for 24 hours to obtain a fermented product. .

division strain name deposit number Example 1 Leuconostoc mesenteroides SST-9962 KCCM12712P Example 2 Lactobacillus paraplantarum SST-9961 KCCM12711P

Example 3: Preparation of fermented antler fermented with Bacillus subtilis

The antler extract obtained by diluting the antler extract of Preparation Example 1 with 4 times the weight of distilled water was sterilized at 121° C., 1.5 atmospheres, and 15 minutes.

In addition, Bacillus subtilis SST-9960 [Accession No.: KCCM 11420P] strain was inoculated in LB liquid medium and then cultured for 2 days at 30 ° C. and activated to prepare a Bacillus subtilis culture.

After that, 1.5 wt% of glucose was added to the deer antler extract, and 1.0 wt% of the Bacillus subtilis culture solution (1×10 ⁸ CFU/mL) was inoculated, followed by aerobic conditions with agitation speed of 100 rpm at 30°C for 3 days. cultured to obtain a fermented product.

Comparative Example: Deer antler hot water extract

The hot water extract of deer antler of Preparation Example 1 was used as a comparative example in the following test.

Test Example 1: Fermentation characteristics of strains in deer antler extract - Change in number of viable cells

In order to confirm the fermentation characteristics of lactic acid bacteria and Bacillus subtilis in the deer antler extract, it was attempted to confirm the change in the number of viable cells at 12 and 24 hours after inoculation.

Specifically, 1.5% by weight of glucose was added to the deer antler extract obtained by diluting the deer antler extract of Preparation Examples 1 to 3 with distilled water 4 times the weight, and the lactic acid bacteria culture solution (1×10 ⁸ CFU/mL) or Bacillus subtilis culture solution. (1×10 ⁸ CFU/mL) was inoculated at 0.5% by weight and then fermented at 37° C. for 24 hours, the number of viable cells was measured, and the results are shown in Table 2 below.

For live cell water, dilute 1 mL of sample in 9 mL of sterile physiological saline, mix well, dilute step by step, take 0.1 mL, use an anaerobic jar (BBL Gas Pak Anaerobic System), and store in anaerobic condition as much as possible and incubate at 37 °C. Then, the number of colonies generated was counted, and the average number of colonies was multiplied by a dilution factor to calculate colonies per culture.

division Number of viable cells (CFU/mL) before fermentation After fermentation (Preparation Example 1) After fermentation (Preparation Example 2) After fermentation (Preparation Example 3) Leukonostok mecenteroides
SST-9962 5.0×10 ⁵ 7.2×10 ⁷ 6.8×10 ⁷ 3.3×10 ⁷ Lactobacillus paraplantarum SST-9961 5.0×10 ⁵ 2.6×10 ⁷ 2.2×10 ⁷ 1.0×10 ⁷ Bacillus subtilis
SST-9960 5.0×10 ⁵ 0.8×10 ⁷ 0.6×10 ⁷ 0.4×10 ⁷

As shown in Table 2, there was little difference between the use of the hot water extract of Preparation Example 1 and the ethanol aqueous extract of Preparation Example 2 as a fermentation substrate. The number of viable cells was slightly less than half the number of viable cells in Preparation Examples 1 and 2.

In addition, it was confirmed that the number of viable cells in the culture medium prepared using the Leuconostoc mesenteroides SST-9962 strain was much (about 3 times) higher than the number of viable cells in the culture medium prepared using the Lactobacillus paraplant arum SST-9961 strain.

Test Example 2: Comparison of growth in antler extracts of lactic acid bacteria and Bacillus subtilis strains

To the lactic acid bacteria and Bacillus subtilis strains of the present invention, it was attempted to compare the growth of the strain in the deer antler extract.

To this end, 1.5% by weight of glucose was added to the antler extract obtained by diluting the antler extract of Preparation Example 1 with distilled water of 2, 4, and 10 times the weight, and the lactic acid bacteria culture solution (1×10 ⁸ CFU/mL) was added to 0.5 After inoculation in wt% and fermented at 37 ° C. for 24 hours, the growth of the strain was measured with a spectrophotometer (Optizen 2120UV, Mecasys co., Ltd., Korea) at 600 nm, and is shown in Table 3 below.

division Viability (OD ₆₀₀ ) before dilution 2x dilution 4-fold dilution 10-fold dilution Example 1 (SST-9962) 0.597 1.112 1.624 0.784 Example 2 (SST-9961) 0.612 1.256 1.823 0.767 Example 3 (SST-9960) 0.609 0.615 0.224 0.110

Looking at Table 3, the growth rate of the Leuconostoc mesenteroides SST-9962 strain and the Lactobacillus paraplant arum SST-9961 strain with respect to the 4-fold diluted antler extract did not differ significantly, but Bacillus against the 4-fold diluted antler extract. The viability of the subtilis strain was relatively low compared to the viability of the SST-9962 strain and the SST-9961 strain.

In addition, in the case of SST-9962 strain and SST-9961 strain, the highest growth was shown in the antler extract diluted 4 times compared to the antler extract diluted 2 times or 10 times before dilution, whereas in the case of Bacillus subtilis, the dilution The highest growth rate was found in the antler extract diluted before or twice.

Test Example 3: Analysis of Cell Viability according to Fermented Deer Antler-1

3-1: Analysis of cell viability in keratinocytes (1)

Cytotoxicity was measured to determine the effect of the fermented antlers prepared in Examples 1 to 3 on the cell viability of HaCaT keratinocytes.

Specifically, under the conditions of 37° C. and 5% CO ₂ , the HaCaT cell line, a human-derived keratinocyte, was treated with DMEM (Dulbecco's Modified) containing 1% penicillin-streptomycin and 10% fetal bovine serum, respectively. Eagle's Medium) medium (Gibco BRL, USA). The HaCaT cell line cultured under the above conditions was aliquoted in a 96-well plate or 10 mL culture dish at 1×10 ⁵ CFU/mL and cultured for 24 hours, then the medium was removed and a fresh medium was added. The control group did not add any inflammatory factors and fermented antlers, and the experimental group added the fermented antlers (10, 50, 100, 200, 500 μg/mL) prepared in Examples 1 to 3 to each of the cells. did it The control group and the experimental group were cultured for 24 hours, and then cell viability was measured.

In order to compare the cell viability according to the concentration of the fermented antler, after recovering each of the prepared cells, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium) solution (0.05) mg/mL concentration) was added, and the reduction reaction was induced while culturing in a cell incubator for 4 hours, then the MTT solution was removed and DMSO was added to completely dissolve formazan crystals. 200 μl of this was put into each well and the absorbance was measured at 570 nm using a microplate reader. Cell viability was calculated as in Equation 1 below, and is shown in Table 4 below.

[Equation 1]

Cell viability (%) = (absorbance of sample treatment group / absorbance of control group) × 100

division Cell viability (%) Sample treatment concentration (㎍/mL) 10 50 100 200 500 Control group (untreated group) 100 Example 1 113.1 132.3 116.5 99.7 94.9 Example 2 108.6 124.5 110.2 101.2 95.4 Example 3 108.7 126.1 109.9 102.4 95.8

As shown in Table 4, it was confirmed that the fermented deer antler according to the embodiment of the present invention did not show cytotoxicity up to a concentration of 500 μg/mL.

3-2: Analysis of cell viability in inflammation-induced keratinocytes (2)

The purpose of this study was to determine the effect of the fermented deer antlers prepared in Examples 1 to 3 on the cell viability of HaCaT keratinocytes induced by inflammation with IFN-γ and TNF-α cytokines.

Specifically, under the conditions of 37° C. and 5% CO ₂ , the HaCaT cell line, a human-derived keratinocyte, was treated with DMEM (Dulbecco's Modified) containing 1% penicillin-streptomycin and 10% fetal bovine serum, respectively. Eagle's Medium) medium (Gibco BRL, USA). The HaCaT cell line cultured under the above conditions was aliquoted in a 96-well plate or 10 mL culture dish at 1×10 ⁵ CFU/mL and cultured for 24 hours, then the medium was removed and a fresh medium was added. The control group was the addition of 10 ng/mL of TNF-α, an inflammatory factor, and the experimental group was carried out with IFN-γ and TNF-α (1: 1 (v/v), 10 ng/mL) to each of the cells. The fermented antler antlers prepared in Examples 1 to 3 (10, 50, 100, 200, 500 μg/mL) were added. After the control and experimental groups were cultured for 24 hours, cell viability was measured in the same manner as in Test Example 3-1, and is shown in Table 5 below.

division Cell viability (%) Sample treatment concentration (㎍/mL) 10 50 100 200 500 Control group (untreated group) 100 Example 1 112.4 96.7 103.4 92.3 93.7 Example 2 107.5 94.6 99.7 89.7 92.5 Example 3 102.2 94.3 96.2 86.1 89.6

As shown in Table 5, it was confirmed that cytotoxicity was not observed in the fermented deer deer antler of the present invention even in cells induced by inflammation by treatment with IFN-γ, an inflammation-inducing substance, up to a concentration of 500 μg/mL. Therefore, even if the inflamed cells are treated with the fermented antler, cytotoxicity does not occur, so it will be very stable.

Test Example 4: Analysis of mRNA expression level of atopic dermatitis-related chemokines-1

Atopic dermatitis is a complex disease with various etiologies and mechanisms, and the pathophysiology is related to dendritic cell-specific T-cell lymphocytes, a network of type 1 and type 2 cytokines, and inflammatory chemokines. Macrophage-derived chemokine (MDC/CCL 22) plays a role in supplying Th2 cells to allergic inflammatory sites during the pathogenesis of atopic dermatitis, and thymus and activation regulated chemokine (TARC) /CCL 17) attracts CCR4+ or CCR8+ cells. In the present invention, the activity of atopic dermatitis was confirmed by measuring the mRNA expression levels of MDC and TARC, which are atopic dermatitis inducing factors that play a role in moving Th2 cells to the site of allergic inflammation, and this was used as an inflammatory index.

To determine whether the MDC and TARC production of the fermented antler of the present invention as a sample was inhibited, mRNA expression levels were measured using RT-PCR. HaCaT cells were inoculated into a 6-well plate at 2.5×10 ⁵ cells per well and 24 hours later, IFN-γ and TNF-α (1 : 1 (v/v)), which are inflammation-inducing substances, were inoculated at 10 ng/ mL, and at the same time treated with various concentrations of fermented deer antlers, and then cultured in a cell incubator at 37° C. supplied with 5% CO ₂ for 24 hours. Total RNA was isolated from cultured cells using TRI (TRIzol) reagent, and complementary DNA was synthesized using cDNA synthesis kit (TOYOBO, Japan). The synthesized cDNA is mixed with the prepared MDC or TARC Primer (BIONEER, Korea), and heated at 60°C for 2 minutes and 95°C for 10 minutes using SYBR Green PCR Master Mix (BIO-RAD, USA) reagent. After the treatment, the gene expression pattern of atopic dermatitis-related factors when an inflammatory factor and antler fermented product were added by reacting under the conditions of a total of 40 cycles for 30 seconds at 95 °C, 30 seconds at 60 °C, and 30 seconds at 72 °C. was analyzed. In the control group, 10 ng/mL of inflammatory factors IFN-γ and TNF-α (1 : 1 (v/v)) were added, and the experimental group was IFN-γ and TNF-α (10 ng/mL) in each of the cells. mL) and the fermented antler antlers prepared in Examples 1 to 3 (10, 50, 100, 200 μg/mL) were added.

Table 6 below shows MDC and TARC expression levels in HaCaT cells treated with samples according to Examples and Comparative Examples, respectively, and compared as relative band densities (% of control).

sample (μg/mL) TARC
(% of control) MDC
(% of control) control 100 100 Example 1 10 8.3±0.3 25.3±1.4 50 6.2±0.4 19.1±1.6 100 4.1±0.7 15.1±2.1 200 4.0±0.5 11.5±0.7 Example 2 10 18.7±1.1 33.8±2.1 50 15.2±0.8 27.6±1.3 100 11.0±0.9 22.5±2.4 200 9.2±0.3 17.9±1.0 Example 3 10 26.2±0.6 43.5±3.2 50 21.3±1.2 35.2±1.6 100 16.2±0.7 28.4±2.2 200 14.3±2.1 21.0±1.3 comparative example 10 56.3±1.3 73.4±2.6 50 50.1±2.7 62.1±1.6 100 43.5±0.5 52.1±0.7 200 34.6±1.8 47.5±1.1

Referring to Table 6, it was confirmed that the fermented deer antler according to the example of the present invention very effectively inhibited the expression of MDC and TARC in keratinocytes compared to the comparative example. This means that the fermented antler of the present invention is remarkably excellent in preventing, improving or treating atopic dermatitis or allergic dermatitis.

Hereinafter, in the case of lactic acid bacteria other than the strains of Examples 1 and 2, it was tried to confirm whether the same effect as above appears.

Examples 4-42: Preparation of fermented deer antler

It was carried out in the same manner as in Examples 1 and 2, except that the lactic acid bacteria of Examples 1 and 2 were replaced with the lactic acid bacteria of Table 7 below to prepare a fermented deer antler. The lactic acid bacteria in Table 7 below are isolated from traditional foods such as yeast and kimchi and commercially available yogurt, and are strains owned by the applicant. The following strain names are an abbreviation of the scientific name of each strain, or an abbreviation of its own number.

division strain name Example 4 Leuconostoc mesenteroides LM-2 Example 5 Leuconostoc mesenteroides LM-3 Example 6 Leuconostoc lactis LL Example 7 Lactobacillus plantarum LP-2 Example 8 Lactobacillus paraplantarum LP-3 Example 9 Lactobacillus plantarum LP-4 Example 10 Lactobacillus plantarum LP-5 Example 11 Lactobacillus sakei LS-1 Example 12 Lactobacillus sakei LS-2 Example 13 Lactobacillus sakei LS-3 Example 14 Lactobacillus sakei LS-4 Example 15 Lactobacillus sakei LS-5 Example 16 Lactobacillus arizonensis LA-1 Example 17 Lactobacillus arizonensis LA-2 Example 18 Lactobacillus johnsonii LJ Example 19 Lactobacillus pentosus LPE Example 20 Lactobacillus brevis LB Example 21 Lactobacillus hayakitensis LH-1 Example 22 Lactobacillus hayakitensis LH-2 Example 23 Lactococcus taiwanensis LT Example 24 Pediococcus pentosaceus PP-1 Example 25 Pediococcus pentosaceus PP-2 Example 26 Pediococcus pentosaceus PP-3 Example 27 Pediococcus pentosaceus PP-4 Example 28 Pediococcus pentosaceus PP-5 Example 29 Pediococcus pentosaceus PP-6 Example 30 Pediococcus pentosaceus PP-7 Example 31 Pediococcus pentosaceus PP-8 Example 32 Pediococcus pentosaceus PP-9 Example 33 Pediococcus pentosaceus PP-10 Example 34 Pediococcus pentosaceus PP-11 Example 35 Weissella confusa WC Example 36 Enterococcus raffinosus ER-1 Example 37 Enterococcus raffinosus ER-2 Example 38 Enterococcus faecalis EF Example 39 Staphylococcus warneri SW Example 40 Staphylococcus xylosus SX Example 41 Bifidobacterium bifidum BB Example 42 Aerococcus urinaeequi AU

Test Example 5: Analysis of mRNA expression level of atopic dermatitis-related chemokines-2

In order to confirm the effect of inhibiting the expression of MDC and TARC in HaCaT cells of lactic acid bacteria other than the strains of Examples 1 and 2, an experiment was conducted in the same manner as in Test Example 4.

Table 8 below shows MDC and TARC expression levels in HaCaT cells after treatment with 100 μg/mL of a sample according to Example, respectively, and compared as relative band density (% of control).

division strain TARC
(% of control) MDC
(% of control) control - 100 100 comparative example - 43.5±0.5 52.1±0.7 Example 4 Leuconostoc mesenteroides LM-2 16.2±1.3 24.4±1.2 Example 7 Lactobacillus plantarum LP-2 20.4±0.9 32.7±0.6 Example 8 Lactobacillus paraplantarum LP-3 17.1±0.6 30.5±0.7 Example 12 Lactobacillus sakei LS-2 24.4±1.4 33.2±1.0 Example 13 Lactobacillus sakei LS-3 26.1±0.8 33.5±0.9 Example 21 Lactobacillus hayakitensis LH-1 30.6±1.7 39.5±1.7 Example 22 Lactobacillus hayakitensis LH-2 28.3±1.4 37.2±0.8 Example 23 Lactococcus taiwanensis LT 27.5±0.6 41.2±1.6 Example 28 Pediococcus pentosaceus PP-5 22.4±0.7 33.6±1.5 Example 33 Pediococcus pentosaceus PP-10 21.6±0.4 34.1±0.3 Example 34 Weissella confusa WC 29.4±1.1 38.0±0.7 Example 40 Staphylococcus xylosus SX 26.7±2.6 36.8±1.3

Looking at Table 8, it was confirmed that the fermented deer antler fermented with lactic acid bacteria other than the strains of Examples 1 and 2 of the present invention further inhibited the expression of MDC and TARC in keratinocytes compared to the Comparative Example. This means that even in the case of fermented deer antlers fermented with lactic acid bacteria other than the strains of Examples 1 and 2, the effect of inhibiting MDC and TARC expression in keratinocytes is remarkably excellent.

Test Example 6: Growth of other lactic acid bacteria in deer antler extract

In order to confirm that other lactic acid bacteria other than the lactic acid bacteria of Examples 1 and 2 also exhibit high growth in the deer antler extract (4-fold dilution), the lactic acid bacteria growth was measured in the same manner as in Test Example 2, and is shown in Table 9 below. .

division strain name Viability (O.D. 600nm) Example 4 Leuconostoc mesenteroides LM-2 1.623 Example 5 Leuconostoc mesenteroides LM-3 1.522 Example 6 Leuconostoc lactis LL 1.526 Example 7 Lactobacillus plantarum LP-2 1.700 Example 8 Lactobacillus paraplantarum LP-3 1.550 Example 9 Lactobacillus plantarum LP-4 1.646 Example 10 Lactobacillus plantarum LP-5 1.544 Example 11 Lactobacillus sakei LS-1 1.695 Example 12 Lactobacillus sakei LS-2 1.584 Example 13 Lactobacillus sakei LS-3 1.676 Example 14 Lactobacillus sakei LS-4 1.584 Example 15 Lactobacillus sakei LS-5 1.637 Example 16 Lactobacillus arizonensis LA-1 1.649 Example 17 Lactobacillus arizonensis LA-2 1.530 Example 18 Lactobacillus johnsonii LJ 1.851 Example 19 Lactobacillus pentosus LPE 1.570 Example 20 Lactobacillus brevis LB 1.564 Example 21 Lactobacillus hayakitensis LH-1 1.595 Example 22 Lactobacillus hayakitensis LH-2 1.563 Example 23 Lactococcus taiwanensis LT 1.643 Example 24 Pediococcus pentosaceus PP-1 1.635 Example 25 Pediococcus pentosaceus PP-2 1.631 Example 26 Pediococcus pentosaceus PP-3 1.618 Example 27 Pediococcus pentosaceus PP-4 1.602 Example 28 Pediococcus pentosaceus PP-5 1.560 Example 29 Pediococcus pentosaceus PP-6 1.560 Example 30 Pediococcus pentosaceus PP-7 1.524 Example 31 Pediococcus pentosaceus PP-8 1.516 Example 32 Pediococcus pentosaceus PP-9 1.548 Example 33 Pediococcus pentosaceus PP-10 1.515 Example 34 Pediococcus pentosaceus PP-11 1.537 Example 35 Weissella confusa WC 1.918 Example 36 Enterococcus raffinosus ER-1 1.792 Example 37 Enterococcus raffinosus ER-2 1.518 Example 38 Enterococcus faecalis EF 1.606 Example 39 Staphylococcus warneri SW 1.710 Example 40 Staphylococcus xylosus SX 1.654 Example 41 Bifidobacterium bifidum BB 1.574 Example 42 Aerococcus urinaeequi AU 1.546

Looking at Table 9, the lactic acid bacteria growth (OD ₆₀₀ ) of the fermented antler was in the range of 1.515 to 1.918, confirming that the lactic acid bacteria had a very high growth in the antler extract.

Test Example 7: Fermentation characteristics of fermented antler ?? pH change

The pH change according to the type of lactic acid bacteria and the culture time is shown in Table 10 below. The pH was measured at room temperature using a pH meter (Seven Compact, Mettler Toledo, Switzerland) after taking 10 mL of the sample.

division strain name 0 hours 24 hours Example 2 Leuconostoc mesenteroides SST-9962 6.42 5.28 Example 6 Leuconostoc lactis LL 6.42 5.40 Example 7 Lactobacillus plantarum LP-2 6.42 5.22 Example 11 Lactobacillus sakei LS-1 6.41 5.38 Example 16 Lactobacillus arizonensis LA-1 6.42 5.47 Example 17 Lactobacillus johnsonii LJ 6.42 5.50 Example 18 Lactobacillus pentosus LPE 6.42 5.46 Example 20 Lactobacillus brevis LB 6.42 5.41 Example 21 Lactobacillus hayakitensis LH-1 6.42 5.51 Example 23 Lactococcus taiwanensis LT 6.41 5.42 Example 28 Pediococcus pentosaceus PP-5 6.42 5.45 Example 35 Weissella confusa WC 6.42 5.49 Example 36 Enterococcus raffinosus ER-1 6.42 5.44 Example 38 Enterococcus faecalis EF 6.42 5.47 Example 39 Staphylococcus warneri SW 6.42 5.64 Example 40 Staphylococcus xylosus SX 6.42 5.58 Example 41 Bifidobacterium bifidum BB 6.42 5.65

Looking at Table 10, when comparing the pH change for each lactic acid bacteria, Leuconostoc mesenteroides SST-9962 and Lactobacillus plantarum LP-2 were significantly decreased compared to other strains.

On the other hand, when the lactic acid bacteria are the same, there was little difference in the pH of the culture medium using the hot water extract of Preparation Example 1, the ethanol extract of Preparation Example 2, and the dilution of the antler water of Preparation Example 3 as substrates, and in Preparation Example 3, the remaining Compared to that, the pH at 24 hours was slightly higher.

<Animal Experiment>

Experimental animals : SPF male NC/Nga mice were used.

Manufacture of atopic dermatitis animal model : After cleaning the back of an 8-week-old male NC/Nga mouse, it was left for 24 hours to heal fine wounds on the skin that may occur during the hair removal process. After 24 hours, 200 μl of 1% DNCB was applied to the depilated skin in the first induction stage. In the second induction stage, which is the second day after the first sensitization, 200 μl of 1% DNCB was applied to the depilated skin. At the challenge stage, which is the fourth day after the second sensitization, 150 μl of 0.4% DNCB was applied to the depilated skin and repeated twice a week (Mon/Thu) for a total of 4 weeks to induce dermatitis.

Sample administration : The test substance was administered orally forcibly from the start of the experiment to the end of the experiment using a sonde for oral administration. The frequency and time of administration were 1 time/day, 7 days/week, 4 weeks, and before 16:00 on the day of administration. The dose was calculated as a dose of 10 ml/kg based on the most recently measured body weight. In order to compare the efficacy, Prednisolone, known as an anti-itch standard, was used. Prednisolone was administered at 5 mg/kg for 4 weeks. The composition of the test group is shown in Table 11 below.

test group Number of animals (number of animals) Dosage G1 normal control group 10 Oral administration of physiological saline G2 AD control 10 Oral administration of physiological saline G3 AD Example 1 10 Sample of Example 1, 200 mg/kg/day oral administration G4 AD Example 2 10 Sample of Example 2, 200 mg/kg/day oral administration G5 AD Example 3 10 Sample of Example 3, 200 mg/kg/day oral administration G6 AD comparative example 10 Sample of Comparative Example, 200 mg/kg/day oral administration G7 Prednisolone 10 Prednisolone, 5 mg/kg/day oral administration * G1: Normal control group, G2: Induction control group, G3~5: Induction test group, G6: Induction comparison group, G7: Positive control group
* AD (atopic dermatitis)

Test Example 8: Measurement of skin thickness

In an atopic dermatitis animal model (NC/Nga mice), the effect of the fermented deer antler according to the present invention on the increase in skin thickness due to skin inflammation was investigated.

After sacrificing the experimental animals of each experimental group and cutting the back skin tissue, the thickness of the back skin tissue of each experimental animal was measured using a digital caliper (Mitutoyo, Japan), and the results are shown in Table 12 below.

division Skin thickness (mm) G1 normal control group 0.75±0.03 G2 AD control 1.96±0.04 G3 AD Example 1 1.09±0.07 G4 AD Example 2 1.18±0.09 G5 AD Example 3 1.27±0.05 G6 AD comparative example 1.62±0.10 G7 Prednisolone 1.33±0.06

As shown in Table 12, it can be seen that the skin thickness of the G3 to G5 experimental groups administered with the fermented antler according to the present invention was significantly reduced compared to G2 (control group) and G6 (comparative group).

Test Example 9: Measurement of the number of scratching

The most symptomatic feature of atopic dermatitis or allergic dermatitis is scratching due to severe itching. This continuous scratching injures the skin, intensifies the inflammatory response and exacerbates the symptoms. Therefore, the method of suppressing such scratching may be an effective treatment method capable of alleviating the symptoms of atopic dermatitis or allergic dermatitis.

At the 3rd week of the experiment, the atopic dermatitis animal model (NC/Nga mice) was acclimatized in a transparent cage for 30 minutes, and the cages of each experimental group were recorded for about 10 minutes using a camcorder, The number of scratching motions was measured and shown in Table 13 below.

division total number of scratches Number of scratches per hour (number of times/min) G1 normal control group 5 0.5 G2 AD control 99 9.9 G3 AD Example 1 12 1.2 G4 AD Example 2 25 2.5 G5 AD Example 3 43 4.3 G6 AD comparative example 79 7.9 G7 Prednisolone 37 3.7

Referring to Table 13, it can be seen that the G3 ~ G5 experimental group administered with the fermented antler according to the present invention significantly reduced the number of scratching compared to G2 (control group) and G6 (comparative group).

Test Example 10: Measurement of blood IgE concentration

The purpose of this study was to examine the effect of fermented deer antler on IgE production in relation to increase in blood IgE due to activation of Th2 cells in peripheral blood, an immunological characteristic of the pathogenesis of atopic dermatitis.

At the 4th week of the experiment, 5 hours after the end of administration of the fermented antler, blood was collected from atopic dermatitis animal models (NC/Nga mice), and then plasma was separated by centrifugation at 10,000 rpm and 4°C for 10 minutes. 100 μL of plasma from each experimental group obtained above was put in a 96-well plate with capture IgE antibody to induce an antigen-antibody reaction at room temperature for 1 hour. At this time, the plate with immunoglobulin E attached was After blocking for 1 hour using 5% skim milk powder in advance, it was washed three times with a washing solution (PBS + 0.1% tween 20). After the antigen-antibody reaction was completed, the cells were washed three more times, and 100 μL of IgE antibody for detection coupled with horseradish peroxidase was put into each well and reacted for 1 hour. After the reaction, after washing with a washing solution 5 times, a chromogenic substrate was injected and reacted, and measured with an ELISA reader (Molecular Devices, USA). For the quantification of each substance, a standard curve was determined by treating each substance by concentration and reacting it to the serum. The amount of IgE contained was calculated. The measured values are shown in Table 14 below.

division IgE (ng/mL) G1 normal control group 11.3 G2 AD control 82.2 G3 AD Example 1 51.3 G4 AD Example 2 55.4 G5 AD Example 3 61.5 G6 AD comparative example 76.5 G7 Prednisolone 62.7

Referring to Table 14, it can be seen that the G3 to G5 experimental groups administered with the fermented antler according to the present invention have superior IgE inhibitory effects in the blood compared to G2 (control) and G6 (comparative). In addition, the G3~G5 experimental group of the present invention had a similar or superior blood IgE inhibitory effect compared to the standard prednisolone.

Hereinafter, a manufacturing example of a food or drug containing the sample of the above example according to the present invention as an active ingredient will be described, but the present invention is not intended to limit the present invention, but merely to describe it in detail. Foods, pharmaceuticals and cosmetics were prepared in Preparation Examples 4 to 6 according to the following composition ingredients and composition ratios with the fermented deer antler having excellent preventive and therapeutic effects on the muscle disease or muscle damage according to a conventional method.

<Production Example>

Preparation Example 4. Preparation of food

Preparation Example 4-1. Manufacturing of health functional food

100 mg of fermented deer antler of Example 1

appropriate amount of vitamin mixture

70 μg vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

0.2 μg of vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

50 μg of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health functional food, but the composition is mixed in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, the granules can be prepared and used in the preparation of a health functional food composition according to a conventional method.

Preparation Example 4-2. Production of functional beverages

100 mg of fermented deer antler of Example 1

1,000 mg citric acid

100 g of oligosaccharides

2 g of plum concentrate

1 g taurine

Add purified water to total 900 mL

After mixing the above ingredients according to the usual health drink manufacturing method, after stirring and heating at 85°C for about 1 hour, the resulting solution is filtered and acquired in a sterilized 2L container, sealed and sterilized, then refrigerated. It is used to prepare the functional beverage composition of the present invention.

Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, demanding country, and use.

Preparation Example 4-3. candy

Sugar 60% by weight, starch syrup 39.8% by weight, and flavoring 0.1% by weight and 0.1% by weight of the fermented antler antler of Example 1 were mixed to prepare a candy in a conventional manner.

Preparation 5. Preparation of pharmaceuticals

Preparation Example 5-1. powder

200 mg of fermented deer antler of Example 1

Lactose 100 mg

talc 10 mg

The above ingredients are mixed and filled in an airtight bag to prepare a powder.

Preparation Example 5-2. refine

500 mg of fermented deer antler of Example 1

100 mg cornstarch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional manufacturing method of tablets.

Preparation Example 5-3. capsules

500 mg of fermented deer antler of Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

0.2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled in a gelatin capsule to prepare a capsule.

Preparation Example 5-4. liquid

100 mg of fermented deer antler of Example 1

10 g isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to a conventional liquid preparation method, each component is added to purified water to dissolve, an appropriate amount of lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 by adding purified water, and then filled in a brown bottle to sterilize to prepare a liquid.

Preparation Example 6. Preparation of cosmetics

Preparation Example 6-1. Manufacture of nourishing lotion (milk lotion)

Nutrient lotion containing the fermented deer antler of the present invention with the ingredients and contents shown in Table 15 below was prepared according to a conventional method.

Ingredients Production Example 6-1
(weight%) Fermented deer antler of Example 1 100 μg/mL squalane 5.0 beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 liquid paraffin 0.5 Caprylic/Capric Triglycerides 5.0 glycerin 3.0 butylene glycol 3.0 propylene glycol 3.0 Carboxyvinyl Polymer 0.1 triethanolamine 0.2 Preservatives, Colors, Flavors appropriate amount Purified water to 100

Preparation Example 6-2. Manufacture of softening lotion (skin lotion)

Nutrient lotion containing the fermented deer antler of the present invention with the ingredients and contents shown in Table 16 below was prepared according to a conventional method.

Ingredients Preparation 6-2
(weight%) Fermented deer antler of Example 1 100 μg/mL glycerin 3.0 butylene glycol 2.0 propylene glycol 2.0 Carboxyvinyl Polymer 0.1 PEG 12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 triethanolamine 0.1 Preservatives, Colors, Flavors appropriate amount Purified water to 100

Preparation Example 6-3. production of nourishing cream

A nutritional cream containing the fermented antler of the present invention with the ingredients and contents shown in Table 17 below was prepared according to a conventional method.

Ingredients Production Example 6-3
(weight%) Fermented deer antler of Example 1 100 μg/mL Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 PEG60 hydrogenated castor oil 2.0 liquid paraffin 10 squalane 5.0 Caprylic/Capric Triglycerides 5.0 glycerin 5.0 butylene glycol 3.0 propylene glycol 3.0 triethanolamine 0.2 antiseptic appropriate amount pigment appropriate amount Spices appropriate amount Purified water to 100

Preparation Example 6-4. manufacture of massage cream

A massage cream comprising the fermented deer antler of the present invention in the amounts and ingredients shown in Table 18 below was prepared according to a conventional method.

Ingredients Preparation 6-4
(weight%) Fermented deer antler of Example 1 100 μg/mL beeswax 10.0 Polysorbate 60 1.5 PEG 60 hydrogenated castor oil 2.0 Sorbitan sesquioleate 0.8 liquid paraffin 40.0 squalane 5.0 Caprylic/Capric Triglycerides 4.0 glycerin 5.0 butylene glycol 3.0 propylene glycol 3.0 triethanolamine 0.2 Preservatives, Colors, Flavors appropriate amount Purified water to 100

Preparation Example 6-5. Preparation of emulsion for mask pack

An emulsion for a mask pack containing the fermented deer antler of the present invention in the amounts and ingredients shown in Table 19 below was prepared according to a conventional method.

Ingredients Preparation 6-5
(weight%) Fermented deer antler of Example 1 100 μg/mL polyvinyl alcohol 13.0 Sodium Carboxymethyl Cellulose 0.2 glycerin 5.0 allantoin 0.1 ethanol 6.0 PEG 12 nonylphenyl ether 0.3 Polysorbate 60 0.3 Preservatives, Colors, Flavors appropriate amount Purified water to 100

Although preferred embodiments of the present invention have been described above, the present invention is not limited thereto, and various modifications can be made within the scope of the technical spirit of the present invention, which also falls within the scope of the appended claims. it is natural

Claims (9)

A food composition for preventing or improving atopic dermatitis or allergic dermatitis comprising fermented antler as an active ingredient. According to claim 1,
The fermented antler is a food composition for preventing or improving atopic dermatitis or allergic dermatitis, characterized in that it is obtained by fermenting an antler extract with lactic acid bacteria or Bacillus subtilis . 3. The method of claim 2,
The antler extract is a food composition for preventing or improving atopic dermatitis or allergic dermatitis, characterized in that the antler is an extract of water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. 3. The method of claim 2,
The lactic acid bacteria are Lactobacillus genus, Bifidobacterium genus, Leuconostok genus, Pediococcus genus, Weissella genus, Staphylococcus genus, Erococcus genus and Enterococcus genus Lactobacillus selected from any one or more lactic acid bacteria A food composition for preventing or improving atopic dermatitis or allergic dermatitis. 5. The method of claim 4,
The lactic acid bacteria is Leuconostoc mesenteroides ( Leuconostoc mesenteroides ), Lactobacillus plantarum ( Lactobacillus plantarum ) and Lactobacillus paraplantarum ( Lactobacillus paraplantarum ) Atopic dermatitis or allergy, characterized in that any one or more lactic acid bacteria selected from A food composition for preventing or improving sexual dermatitis. According to claim 1,
The composition is a food composition for preventing or improving atopic dermatitis or allergic dermatitis, characterized in that it suppresses the expression of MDC (Marcrophage-derived chemokine) and TARC (Thymus and activation regulated chemokine). According to claim 1,
The composition is a food composition for preventing or improving atopic dermatitis or allergic dermatitis, characterized in that it reduces the amount of IgG in serum. A pharmaceutical composition for preventing or treating atopic dermatitis or allergic dermatitis comprising fermented antler as an active ingredient. A cosmetic composition for preventing or improving atopic dermatitis or allergic dermatitis comprising fermented antler as an active ingredient.