KR20200120556A - Composition for improving, preventing or treating skin disease comprising Thymus plant extract - Google Patents

Abstract

The present invention relates to a composition for alleviating, treating, or preventing allergic or inflammatory skin diseases. More specifically, the composition contains a Thymus vulgaris extract, or the Thymus vulgaris extract and an Echinacea purpurea juice as an active component to reduce an ear edema in an inflammatory skin disease animal model, reduce the levels of total leukocytes, neutrophils, eosinophils, and blood IgE and the like in blood tests, and thus to reduce histopathologically, the levels of inflammatory cell infiltration and mast cell infiltration in the epidermis, thereby being able to be used as a pharmaceutical composition for treating or preventing allergic or inflammatory skin diseases, a pharmaceutical composition for use in animals for treating or preventing allergic or inflammatory skin diseases, a food composition for alleviating allergic or inflammatory skin diseases, or a feed composition for alleviating allergic or inflammatory skin diseases.

Description

Composition for improving, preventing or treating skin disease comprising Thymus plant extract}

The present invention relates to a composition for improving, treating or preventing skin diseases, in particular allergic or inflammatory skin diseases, and more particularly, allergic or inflammatory skin comprising thyme extract, or thyme extract and Jasper Cheon Inguk juice as an active ingredient It relates to a pharmaceutical composition for the treatment or prevention of diseases, a pharmaceutical composition for animals for the treatment or prevention of allergic or inflammatory skin diseases, a food composition for improving allergic or inflammatory skin diseases, or a feed composition for improving allergic or inflammatory skin diseases.

Human skin consists of the dermis and epidermis. In particular, the epidermis, located at the outermost part of the skin, protects against various stimuli from the outside, such as physicochemical stimuli such as chemicals, air pollutants, dry environments, and ultraviolet rays, and prevents excessive release of body moisture through the skin. It performs a protective function of preventing.

Among the epidermis, the outermost stratum corneum is formed from keratinocytes, and consists of keratinocytes that have been differentiated and the lipid layer surrounding them. Keratinocytes are eliminated from the skin after a certain period of time, and new keratinocytes raised from the lowermost layer of the epidermis take over their function.These repetitive processes of change are called epidermis differentiation or keratinization. . In the process of keratinization, keratinocytes form a stratum corneum while generating natural moisturizing factor (NMF) and intercellular lipids, making the stratum corneum firm and flexible, acting as a barrier to the outside as a skin barrier. You will have the function.

The stratum corneum may easily lose its function due to excessive washing, lifestyle factors such as bathing, environmental factors such as dry air pollutants, and endogenous diseases such as atopic skin or senile skin. In fact, in modern times, as harmful factors to the skin are gradually increasing, the rate of formation and dropout of the stratum corneum is slowed due to changes in dietary habits, and the amount of moisturizing factors and lipids in the stratum corneum decreases due to the decline in the function of keratinocytes. There is an increasing trend of people with skin whose stratum corneum does not function as a normal skin barrier.

In particular, atopic dermatitis is an allergic or inflammatory skin disease that has a high risk as the prevalence of atopic dermatitis increases rapidly in recent years.It is estimated that it is related to genetic factors and immune system deficiency, but the exact cause has not yet been identified. It is expected to be somewhat alleviated through improvement of diet, but there is no fundamental treatment method.

It has been found that atopic dermatitis causes IgE to be produced in mast cells in the process of removing allergens that have come into contact with or in the body, and if it comes into contact with or comes in with the same allergens, the human body becomes hypersensitive to allergens. As a result, atopic dermatitis develops by generating histamine.

As the treatment for atopic dermatitis, steroid topical drugs, antihistamines, calcineurin inhibitors, antibiotics, and ultraviolet therapy have been mainly used.However, in the case of long-term use, skin atrophy, capillary dilation, etc., as steroids are absorbed through the skin layer and into blood vessels. A reaction has occurred and there is a limitation due to various side effects such as a risk of inducing adrenal suppression, so there are relatively few side effects and interest in new treatments capable of treating atopy is increasing.

On the other hand, thyme ( Thymus vulgaris ) is a short woody plant of the Lamiaceae family, distributed in Europe and Africa, and is known to have anti-inflammatory, expectorant, antibacterial, bad breath, soothing and sweating effects as a folk remedy.

Regarding Thymus vulgaris , a 2016 paper by Rashid Ismaili et al. discloses that it is effective in treating contact dermatitis by applying essential oil of thyme to the skin, and Japanese Patent Publication No. 2015-107953 discloses dermal extract and thyme extract. And bathroom products mixed with rooibos tea extract have been disclosed to moisturize and protect skin barriers, but the atopic treatment effect of oral administration of thyme extract is not disclosed at all.

In addition, Echinacea purpurea is a perennial plant of the Asteraceae family, native to North America, and is widely cultivated for ornamental purposes all over the world because of its gorgeous flowers, and is also used as a remedy for colds and various viral infections by enhancing immunity.

Regarding Echinacea purpurea , Korean Patent Publication No. 2010-0074688 discloses that tissue cultured Echinacea adventitious root extract can be used as a cosmetic composition because it has a skin moisturizing effect without causing skin irritation. No disclosure has been made on the effect of oral administration of Jajucheonin-guk juice to treat atopy.

Japanese Patent Publication No. 2015-107953 Korean Patent Publication No. 2010-0074688

Rachid Ismaili et al., International Journal of Innovation and Applied Studies 2016, 14(1): p.113-120 Attila Olahar et al., Journal of Dermatological Science 2017, 88: p.67-77

The problem to be solved by the present invention is to provide a pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases comprising thyme extract, or thyme extract, and Japonica Cheonin Guk juice as an active ingredient.

Another problem to be solved by the present invention is to provide a pharmaceutical composition for animals for the treatment or prevention of allergic or inflammatory skin diseases comprising thyme extract, or thyme extract, and Japonica Cheonin Guk juice as an active ingredient.

Another problem to be solved by the present invention is to provide a food composition for improving allergic or inflammatory skin diseases comprising thyme extract, or thyme extract, and Japonica Cheonin Guk juice as an active ingredient.

Another problem to be solved by the present invention is to provide a feed composition for improving allergic or inflammatory skin diseases comprising thyme extract, or thyme extract, and Japonica Cheonin Guk juice as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for treating or preventing allergic or inflammatory skin diseases comprising thyme extract as an active ingredient.

According to an embodiment of the present invention, the allergic or inflammatory skin disease is skin inflammation, eczema, contact dermatitis, atopic dermatitis, seborrheic dermatitis, chronic lichen planus, intermittent dermatitis, deprived dermatitis, papular urticaria, psoriasis, psoriatic arthritis. , Sun dermatitis, sunburn and acne may be any one of the skin diseases selected from.

According to an embodiment of the present invention, the thyme may be a stem, leaf, or above-ground outpost of thyme.

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

According to an embodiment of the present invention, the pharmaceutical composition may further include a juice juice of Jajucheonin-guk.

According to an embodiment of the present invention, the mixing ratio of the thyme extract and the juice of Jajucheonin Guk may be a solid content-based mixing ratio of 1:99 to 99:1 by weight.

According to an embodiment of the present invention, the self-made heaven-in-guk may be a stem, leaf, or above-ground outpost of the self-made heaven-in-guk.

According to an embodiment of the present invention, the pharmaceutical composition may be an oral formulation.

According to an embodiment of the present invention, the oral preparation may be a powder, granule, tablet, capsule, troche, suspension, emulsion, syrup or aerosol.

In addition, the present invention provides an animal pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases comprising thyme extract as an active ingredient.

According to an embodiment of the present invention, the pharmaceutical composition for animals may additionally contain a juice juice of Jasper Cheon In Guk.

In addition, the present invention provides a food composition for improving allergic or inflammatory skin diseases comprising thyme extract as an active ingredient.

According to an embodiment of the present invention, the food composition may additionally include a juice juice of Jajucheoninguk.

According to an embodiment of the present invention, the food composition may be a powder, granule, tablet, capsule, pill, extract, jelly formulation, tea bag formulation, or beverage formulation.

According to an embodiment of the present invention, the food composition may be a health functional food for improving allergic or inflammatory skin diseases.

In addition, the present invention provides a feed composition for improving allergic or inflammatory skin diseases comprising thyme extract as an active ingredient.

According to an embodiment of the present invention, the feed composition may additionally contain a juice juice of Jinghuicheoninguk.

In addition, the present invention provides a method for treating allergic or inflammatory skin diseases in which the composition is orally administered to humans or animals other than humans.

In addition, the present invention provides a novel use of a thyme extract, or thyme extract, and Jasper Cheonin Guk juice for the manufacture of a medicine for the treatment of allergic or inflammatory skin diseases, or an animal medicine.

The composition for the improvement, treatment or prevention of allergic or inflammatory skin diseases comprising thyme extract, or thyme extract and Japonica Cheonguk juice of the present invention as active ingredients, reduces swelling of the ears of inflammatory skin diseases animal models, Leukocytes, neutrophils, eosinophils, reduce the level of IgE in the blood, and histopathologically reduce the level of infiltration of inflammatory cells in the epidermis and the level of infiltration of mast cells, so a pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases, allergic or inflammatory skin It may be used as an animal pharmaceutical composition for treating or preventing diseases, a food composition for improving allergic or inflammatory skin diseases, or a feed composition for improving allergic or inflammatory skin diseases.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention orally administer thyme extract, thyme extract, and Jasper Cheonin Guk juice in an animal model of an allergic or inflammatory skin disease, and the level and tissue of total leukocytes, neutrophils, eosinophils, blood IgE, etc. Pathologically, the level of inflammatory cell infiltration and mast cell infiltration in the epidermis were evaluated. As a result, it was confirmed that the thyme extract, or thyme extract, and the oral administration of Jajucheonin-guk juice, had a remarkable therapeutic effect on allergic or inflammatory skin diseases.

The present invention relates to a pharmaceutical composition for treating or preventing allergic or inflammatory skin diseases comprising thyme extract as an active ingredient.

In addition, the present invention relates to an animal pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases comprising thyme extract as an active ingredient.

The thyme (Thyme) is known as thyme, thymus vulgaris ( Thymus vulgaris ), Timus zygis ( Thymus zygis ) or a mixture thereof, and is distinguished from thyme ( Thymus quinquecostatus ). Thyme has remarkably superior anti-inflammatory activity compared to thyme, so it has a remarkably superior treatment effect for allergic or inflammatory skin diseases than thyme.

The thyme may be an extract of the leaves, stems of thyme, or an above-ground outpost containing the same, but the extract of the above-ground outpost is excellent in treating allergic or inflammatory skin diseases by oral administration.

The thyme extract may be an extract using water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

When the water is suitable for food production, there is no need to specifically limit it, but, for example, ground water, purified water, distilled water, deionized water, and the like may be used.

The alcohol having 1 to 4 carbon atoms is not particularly limited, but, for example, methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol or normal-butanol may be used, preferably ethanol.

The mixed solvent is not particularly limited, but for example, in the case of a mixed solvent of water and ethanol, 5 to 95% by weight aqueous ethanol solution, 10 to 90% by weight aqueous ethanol solution, 20 to 80% by weight aqueous ethanol solution, 30 to 70% by weight An aqueous ethanol solution can be used.

The preparation of the water extract is not particularly limited, but can be prepared by extracting the time with water at 10 to 100° C. for 2 to 60 hours.

The preparation of the alcohol extract or the extract of a mixed solvent of water and alcohol is not particularly limited, but may be prepared by extracting the time at 20 to 60° C. for 2 to 48 hours with an aqueous ethanol solution of 30 to 70% by weight. have.

The extract with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof includes a fraction obtained by re-fractionating the extract with water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof with an organic solvent. The organic solvent may be one or more organic solvents selected from alcohol having 1 to 4 carbon atoms, hexane, acetone, ethyl acetate, chloroform and diethyl ether, and preferably hexane or ethyl acetate.

The term'extract' as used in the present invention is an extract obtained by extracting components contained in Thyme using the solvent, a fraction fractionated therefrom, a concentrate obtained by additionally concentrating these extracts or fractions, and purifying or separating the extract. Water is also included, and the extract, fraction, concentrate, or purified product is used in a sense to include a dried product or a powder obtained by pulverizing it.

For the preparation of the purified product, various additionally carried out, such as passing through an ultrafiltration membrane having a molecular weight cut-off value, or separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity) Purification methods can be added.

The term'allergic or inflammatory skin disease' used in the present invention may include, without limitation, skin diseases that moisturize an allergic or inflammatory reaction occurring in the skin, for example, skin inflammation, eczema, contact dermatitis, atopic dermatitis. , Seborrheic dermatitis, chronic lichen planus, interstitial, deprived dermatitis, papular urticaria, psoriasis, psoriatic arthritis, sun dermatitis, sunburn and acne, preferably any one skin disease, preferably chronic refractory allergic or It may be atopic dermatitis, an inflammatory skin disease.

When the thyme extract is mixed with the juice of Jajucheonin-guk and administered orally as a complex, the synergistic effect of the treatment of allergic or inflammatory skin diseases is remarkably superior compared to the case of oral administration of the thyme extract alone.

When administered orally in combination with the thyme extract, the Jagalcheonin Guk juice is compared to an aqueous extract of Jagalcheonin Guk using water, alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, or a fat-soluble extract fractionated with hexane, ethyl acetate, etc. It is remarkably effective in treating allergic or inflammatory skin diseases.

The juice of the Jasper Cheonin Guk may be an extract of the leaves, stems of Echinacea purpurea , or an above-ground outpost containing the same, but the effect of reducing allergic or inflammatory skin diseases by oral administration is excellent.

The term'juice liquor' as used in the present invention refers to a juice obtained by compressing, pulverizing and compressing the stem, leaves, or above-ground outposts, fractions fractionated from them, concentrates obtained by additionally concentrating these juices or fractions, Purified or separated purified product is also included, and the juice, fraction, concentrate, or purified product is dried or dried or pulverized.

In order to prepare 1 part by weight of the Jasper Cheonin Guk juice powder, about 20 to 100 parts by weight, preferably 30 to 60 parts by weight of undried Jasper Cheoninguk juice, and 1 part by weight of the Jasper Cheon In Guk juice powder 10 to 50 parts by weight of the liquid, preferably 15 to 30 parts by weight, are required.

The mixing ratio based on the solid content of the thyme extract and the Jasper Cheonin Guk juice is from 1: 99 to 99: 1 weight ratio, preferably from 5: 95 to 95: 5 weight ratio, and does not need to be particularly limited, but 10: 90 weight ratio, 20 : 80 weight ratio, 30:70 weight ratio, 40:60 weight ratio, 50:50 weight ratio, 60:40 weight ratio, 70:30 weight ratio, 80:20 weight ratio, 90:10 weight ratio, or any range therebetween.

The'pharmaceutical composition','pharmaceutical','veterinary pharmaceutical composition', or'veterinary medicine' is an active ingredient, in addition to thyme extract, thyme extract, and juice juice, suitable for conventional use in the manufacture of pharmaceutical compositions, etc. It may further include a carrier, excipient and diluent.

The'carrier' is a compound that facilitates the addition of the compound into cells or tissues. The'excipient' is a compound added to provide an appropriate form to the active ingredient to be formulated or to increase the amount for convenient use. The'diluent' is a compound that is diluted in water to dissolve the compound as well as stabilize the biologically active form of the target compound.

The carrier, excipient, and diluent are not particularly limited, but for example, lactose, glucose, sugar, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

The amount of use of the pharmaceutical composition, medicament, veterinary pharmaceutical composition or veterinary medicament may vary depending on the age, sex, and weight of the patient or the animal to be treated, and above all, the condition of the subject to be treated, a specific category of the disease to be treated, or It will depend on the type, route of administration and the nature of the therapeutic agent used.

The pharmaceutical composition, medicament, veterinary pharmaceutical composition or veterinary medicament is appropriately selected according to the absorption of the active ingredient in the body, the excretion rate, the age and weight, sex and condition of the patient or the animal to be treated, and the severity of the disease to be treated. , In general, it is preferred to administer 10 to 5,000 mg per day, preferably 50 to 4,000 mg, more preferably 100 to 3,000 mg, and most preferably 200 to 2,000 mg per day. The unit dosage form formulation thus formulated may be administered several times at regular time intervals as needed.

The pharmaceutical composition, medicament, veterinary pharmaceutical composition, or veterinary medicament may be individually administered as a prophylactic or therapeutic agent, or may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent.

The pharmaceutical composition, medicament, veterinary pharmaceutical composition, or veterinary medicament is formulated and used in oral preparations such as powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, aerosols, etc., according to a conventional method. I can. In the case of formulation, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, and the like, and such solid preparations include at least one excipient for the compound, such as starch, calcium carbonate, sugar or lactose, gelatin. It can be prepared by mixing and the like. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. .

The treatment method of the allergic or inflammatory skin disease is to orally administer the composition to humans or non-human animals, especially mammals, for example, oral administration of the composition to an individual to be treated with an allergic or inflammatory skin disease. It is to administer.

The dosage, administration method, and number of administrations for the treatment may refer to the dosage, administration method, and number of administrations of the pharmaceutical composition, medicine, veterinary pharmaceutical composition or animal medicine.

The present invention relates to a food composition for improving allergic or inflammatory skin diseases comprising thyme extract, or thyme extract, and Japonica Cheonin Guk juice as an active ingredient.

The'food composition' is an active ingredient, in addition to thyme extract, thyme extract, and Japonica Cheoninguk juice, as well as food ingredients and foods that can be used as foods described in the standards and standards of foods commonly used in food manufacturing ('Food Code'). It may contain food additives described in the additive code.

The food composition is not particularly limited, but may include, for example, carbohydrates and flavoring agents. The carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sugar, lactose, and the like; Oligosaccharides or polysaccharides such as dextrin, starch syrup, cyclodextrin, and the like; Sugar alcohols such as xylitol, sorbitol, erythritol, and the like can be used. The flavoring agent may be a natural flavoring agent [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)] and a synthetic flavoring agent (saccharin, aspartame, etc.).

In the case of preparing a food composition using the thyme extract, or thyme extract and Jasmine Cheonin Guk juice as an active ingredient, the thyme extract, or thyme extract and Jasmine Cheonin Guk juice should be specifically limited as long as the content exhibits an allergic or inflammatory skin disease improvement effect. There is no, for example, 0.1 to 99% by weight, 0.5 to 95% by weight, 1 to 90% by weight, 2 to 80% by weight, 3 to 70% by weight, 4 to 60% by weight, may be included in 5 to 50% by weight have.

The thyme extract, or thyme extract, and the juice of Jasper Cheonin Guk, which are active ingredients in the food composition, vary depending on the condition, weight, and the presence or absence of disease, and the duration of the disease, but may be appropriately selected by a person skilled in the art. For example, it may be 10 to 5,000 mg per day, preferably 50 to 4,000 mg, more preferably 100 to 3,000 mg, most preferably 200 to 2,000 mg, based on the daily dose, and the number of administration is particularly Although it does not need to be limited, it can be adjusted by a person skilled in the art within the range of 3 times a day to once a week. In the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, it may be less than the above range.

The food composition is not particularly limited, but may be, for example, a powder, a granule, a tablet, a capsule, a pill, an extract, a jelly formulation, a tea bag formulation, or a beverage formulation.

In addition, the thyme extract, or thyme extract and Jasper Cheonin Guk juice may be added in order to impart the function of improving allergic or inflammatory skin diseases to general food. Foods that can be added do not need to be particularly limited, but for example, confectionery, bread or rice cakes, processed cocoa products or chocolates, meat, etc. as exemplified in the food standards and standards ('Food Code') according to Article 7 of the Food Sanitation Act. Egg processed products, processed fish products, tofu or jelly, noodles, tea, coffee, beverages, special purpose foods, pastes, seasoned foods, dressings, kimchi, salted fish, pickles, stewed foods, alcoholic beverages, raisins, and other foods. Can be. In addition, it may be added to dairy products, processed meat products and packaged meats, and egg products exemplified in the processing standards and ingredient specifications of livestock products according to Article 4 of the Livestock Hygiene Management Act ('Livestock Products Code').

On the other hand, the thyme extract, or thyme extract, and a food composition comprising the juice of Jasper Cheonin Guk as active ingredients is a health functional food for improving allergic or inflammatory skin diseases, for example, a health functional food for improving atopic dermatitis, "for alleviating hyperimmune reactions. It can be used as a helpful health functional food" or as a "health functional food that helps improve skin conditions due to immune hypersensitivity reactions".

The'health functional food' refers to a food manufactured (including processing) in accordance with legal standards using raw materials or ingredients having useful functions for the human body (Article 3, No. 1 of the Health Functional Food Act). The'health functional food' may differ in terms or ranges from country to country, but'Dietary Supplement' in the United States,'Food Supplemnet' in Europe,'Health Functional Food' in Japan or ' It may correspond to'Food for Special Health Use (FoSHU)' and'Health Food' in China.

The food composition or health functional food may additionally contain food additives, and the suitability as a food additive shall be determined according to the standards and standards for the relevant item in accordance with the general rules and general test methods of the'Food Additive Code' unless otherwise specified. Follows.

In addition, the health functional food includes thyme extract, or thyme extract, and "health functional food that helps alleviate hyperimmune reactions" or "health functional food that helps improve skin condition due to immune hypersensitivity reactions" together with thyme extract, or thyme extract, and Jajucheoninguk juice. As a raw material notified as a'functional raw material' or individually recognized as a'functional raw material' used in the process, Enterococcus feacalis heat-treated dry powder, guava leaf extract, etc., stalk extract, leaflet extract, picao preto powder, etc., synthetic PLAG, L. sakei Probio 65, gamma-linolenic acid-containing oil, fruit and vegetable derived lactic acid bacteria ( L. plantarum CJLP133), probiotics ATP and other health functional food materials related to alleviation of hyperimmune reactions can be used in combination.

The present invention relates to a feed composition for improving allergic or inflammatory skin diseases comprising thyme extract, or thyme extract, and Japonica Cheonin Guk juice as an active ingredient.

The'feed composition' is a food ingredient that can be used as a food described in the standards and standards of food ('Food Code'), in addition to thyme extract, thyme extract, and Jasmine Cheonin Guk juice as an active ingredient, food additives listed in the Code of Food Additives. Even if it is not a food raw material or food additive that can be used and can be used as a food, raw materials that fall within the range of sweetened feed in Attached Table 1 of'Standards and Specifications for Feed, etc.', and raw materials that fall within the range of supplementary feeds in Attached Table 2 may be used. .

The'feed composition' may be an extractant among auxiliary feeds according to'standards and standards for feed, etc.', and may be a blended feed including the auxiliary feed.

In the case of preparing a feed composition using the thyme extract, or thyme extract and Jasmine Cheonin Guk juice as active ingredients, the thyme extract, or thyme extract and Jasmine Cheonin Guk juice should be specifically limited as long as the content exhibits allergic or inflammatory skin disease improvement efficacy. There is no, for example, 0.1 to 99% by weight, 0.5 to 95% by weight, 1 to 90% by weight, 2 to 80% by weight, 3 to 70% by weight, 4 to 60% by weight, may be included in 5 to 50% by weight have.

In the feed composition, thyme extract, thyme extract or thyme extract, and the juice of Jagal Cheonin Guk vary depending on the condition, weight, and the presence or absence of disease, and the duration of the disease, but may be appropriately selected by a person skilled in the art. For example, 1 to 5,000 mg, preferably 5 to 2,000 mg, more preferably 10 to 1,000 mg, even more preferably 20 to 800 mg, most preferably 50 to 500 mg, based on daily dosage It may be, the number of administration is not particularly limited, but can be adjusted by a person skilled in the art within the range of 3 times a day to once a week. In the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, it may be less than the above range.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. However, these examples are for describing 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 thereto.

Preparation Example 1: Thyme water extract (TVWE) powder

Fingelberg's "Model No.: 0 196 120; Thyme dry extract" was purchased and used as thyme water extract (TVWE) powder. The thyme water extract (TVWE) powder was a water extract of thyme outpost, and the solid content of thyme water extract was 70% by weight.

Preparation Example 2: Thyme ethanol extract (TVEE) powder

Fingelberg's "Model Number: 0 196 360; Thyme dry extract" was purchased and used as thyme ethanol extract (TVEE) powder. The thyme ethanol extract (TVEE) powder was a 70 wt% ethanol aqueous solution extract of thyme outpost, and the thyme ethanol extract solid content was 80 wt%.

Preparation Example 3: Jajucheoninguk juice (EPJ) powder

Fingelberg's "Model No.: 0 347 318; Echinacea dried pressed juice (95% native)" was purchased and used as an EPJ powder. The Judge Cheonin Guk juice (EPJ) powder was dried from the juice obtained by extracting the outpost, and the JJJJJJ juice juice had a solid content of 95% by weight.

Preparation Example 4: Preparation of thyme extract and juice juice mixture (TEmix1) powder

The thyme water extract (TVWE) powder of Preparation Example 1 and the EPJ powder of Preparation Example 3 were mixed in a ratio of 1:1 by weight based on the solid content of the thyme water extract and the Jagal Cheonin Guk juice, A liquid mixture (TEmix1) powder was prepared.

Preparation Example 5: Preparation of thyme extract and juice juice mixture (TEmix2) powder

The thyme ethanol extract (TVEE) powder of Preparation Example 2 and the EPJ powder of Preparation Example 3 were mixed in a ratio of 1:1 by weight based on the solid content of the thyme ethanol extract and the Jagal Cheonin Guk juice, A liquid mixture (TEmix2) powder was prepared.

Experimental Example 1

1. Preparation of experimental animals and samples

As a positive control group 1,'Styria extract', which is notified as a'functional raw material' used for "health functional food that helps alleviate hyperimmune reaction," was purchased and used. The edible extract was a brown powder, and the solid content of the extract was 41% by weight.

'Dexamethasone Tablet (DEX)' was purchased and used as a positive control group 2. The dexamethasone tablet is a white round tablet containing 0.5 mg of dexamethasone per sugar.

Experimental animals were supplied with Nc/Nga mice (4 weeks old, male) from Japan SLC, Inc., Japan and used. NC/Nga mice show a pattern of atopic dermatitis accompanied by overproduction of IgE, clinically and histologically, very similar to the lesions of human atopic dermatitis, and there are many cases that have been used as atopic dermatitis animal models. I chose.

Sufficient supply of solid feed and water, temperature 23±2 ℃, relative humidity 55±10%, ventilation frequency 10 to 20 times/hr, 12 hours to 12 hours (light-dark cycle) and illuminance 150 to 300 Lux. Breed.

Before the start of breeding, the auricle and the back of the neck of NC/Nga mice were depilated with a razor, and then, an appropriate amount of a depilatory agent was applied to remove the hair. Before administration of the test substance, atopic dermatitis-inducing reagent (Biostir AD, Biostir Inc., Japan) was applied to the back of the auricle, neck, and forelimbs 4 times in total on days 1, 5, 8 and 12 to induce atopic dermatitis. Evenly applied. On the 14th day, the skin clinical index was evaluated and distributed in a random method so that the average level of each group was distributed as uniformly as possible according to the ranked levels of atopic dermatitis.

In order to continuously induce atopic dermatitis even after the start of administration of the test substance, atopic dermatitis-inducing reagent was applied to the auricle, neck, and forelimb locations a total of 10 times on days 15, 19, 22, 26, 29, 33, 36, 40, 43 and 47. It was evenly applied to the back of the body.

All animal testing procedures were performed in compliance with NIH (National Institutes of Health)'s Principle of Laboratory Animal Care.

As shown in Table 1 below, group 1 of them induces atopic dermatitis for 2 weeks in the normal control group, and the remaining 7 groups induce atopic dermatitis for 2 weeks, and then start administering the test substance and induce atopic dermatitis for 5 weeks. Groups 3 to 1-6 are based on the solid content of thyme alone extract or thyme extract and Jajucheoninguk juice mixture excluding excipients, and groups 1-7 are based on the solid content of the extract of'Scythe extract' 200 mg/kg, groups 1-8 were orally administered 2 mg/kg based on the active ingredient of'dexamethasone' once a day based on a 10 mL/kg dose. The normal control group and the negative control group were orally administered 10 mL/kg of physiological saline as a vehicle.

division Explanation Group 1-1 Normal control, does not cause atopic dermatitis, physiological saline Group 1-2 Negative control, atopic dermatitis induced, physiological saline Group 1-3 Inducing atopic dermatitis, Preparation Example 1 TVWE 200 mg/kg Group 1-4 Inducing atopic dermatitis, Preparation Example 2 TVEE 200 mg/kg Group 1-5 Inducing atopic dermatitis, Preparation Example 4 TEmix1 200 mg/kg Group 1-6 Inducing atopic dermatitis, Preparation Example 5 TEmix2 200 mg/kg Group 1-7 Positive control group 1, induces atopic dermatitis, Styria extract 200 mg/kg Group 1-8 Positive control group 2, induced atopic dermatitis, dexamethasone 2 mg/kg

2. Weight measurement result

The body weights (g) on days 1, 14, 28, 42 and 49 of the onset of atopic dermatitis induction were measured and shown in Tables 2 and 3.

division Group 1-1 Group 1-2 Group 1-3 Group 1-4 Day 1 22.34±1.52 22.10±0.77 21.02±1.47 22.01±1.38 Day 14 25.90±1.85 25.57±0.95 24.73±1.58 25.44±1.34 Day 28 27.65±1.77 26.25±1.31 24.40±1.47 ^** 25.24±1.31 ^* Day 42 28.44±2.02 26.55±1.17 25.34±2.38 ^* 26.49±1.47 Day 49 29.98±2.29 27.67±1.23 25.90±2.40 ^** 27.44±1.85

division Group 1-5 Group 1-6 Group 1-7 Group 1-8 Day 1 21.80±1.01 21.40±1.53 21.22±1.64 21.24±0.80 Day 14 24.55±1.14 24.64±0.94 24.47±2.02 24.54±0.99 Day 28 24.48±1.65 ^** 25.10±1.30 ^* 24.81±2.51 ^* 23.26±0.88 ^***,## Day 42 25.50±1.61 ^* 26.30±1.31 25.79±2.54 23.37±1.11 ^***,# Day 49 26.00±1.84 ^** 27.42±1.34 26.33±2.46 ^** 24.19±1.32 ^***,##

Significant differences between the normal control group of group 1-1 and group 1-2 to group 1-8 were * p<0.05, ** p<0.01, *** p<0.001; Significant differences between the negative control group 1-2 and groups 1-3 to 1-8 were # p<0.05, ## p<0.01

On the 14th day of the onset of atopic dermatitis induction, no statistically significant difference in body weight was observed across all test groups.

On the 28th day of the onset of atopic dermatitis (day 14 of test substance administration), the body weights of all test substance-administered groups 1-3 to 1-8 were significantly lower than that of the normal control group 1-1 (p<0.01). , p<0.05, p<0.01, p<0.05, p<0.05 and p<0.001), in particular, the body weight of group 1-8 administered with dexamethasone was significantly lower than that of the negative control group 1-2 (p <0.01).

On the 42nd day of the onset of atopic dermatitis (day 28 of test substance administration), the body weights of groups 1-3, 1-5, and 1-8 were significantly lower than that of the normal control group 1-1 (p< 0.05, p<0.05 and p<0.001), the body weight of groups 1-8 administered with dexamethasone was significantly lower than that of the negative control group of group 1-2 (p<0.01).

On the 49th day of the onset of atopic dermatitis (day 35 of test substance administration), the body weights of groups 1-3, 1-5, 1-7, and 1-8 were compared to the normal control group 1-1. It was significantly lower (p<0.01, p<0.01, p<0.01, and p<0.001), and the body weight of groups 1-8 receiving dexamethasone was significantly lower than that of the negative control group 1-2 (p<0.01). ).

Significant weight loss in groups 1-8, which appeared from day 28 to day 49 of the onset of atopic dermatitis, was evaluated as a side effect of systemic administration of immunosuppressants, compared to group 1-1 observed in other test substance administration groups. Since no significant difference was observed compared to group 1-2 in most of the dates, it was evaluated not due to the administration of the test substance.

3. Ear edema measurement result

Initiation of atopic dermatitis induction 1, 14, 28, 42, and 49 Primary ear thickness (mm) was measured with a caliper to confirm the occurrence of ear edema, and are shown in Tables 4 and 5.

division Group 1-1 Group 1-2 Group 1-3 Group 1-4 Day 1 0.32±0.02 0.31±0.01 0.33±0.01 ^# 0.31±0.01 Day 14 0.33±0.02 0.40±0.06 0.47±0.11 0.46±0.18 Day 28 0.33±0.01 0.70±0.12 ^*** 0.59±0.15 ^** 0.67±0.17 ^*** Day 42 0.33±0.01 0.89±0.09 ^*** 0.69±0.14 ^*** 0.80±0.12 ^*** Day 49 0.33±0.01 0.93±0.11 ^*** 0.73±0.19 ^*** 0.84±0.10 ^***

division Group 1-5 Group 1-6 Group 1-7 Group 1-8 Day 1 0.31±0.01 0.32±0.01 0.33±0.01 0.32±0.02 Day 14 0.46±0.10 0.45±0.09 0.42±0.09 0.47±0.17 Day 28 0.71±0.11 ^*** 0.61±0.16 ^** 0.61±0.09 ^** 0.45±0.11 ^## Day 42 0.82±0.21 ^*** 0.72±0.19 ^*** 0.73±0.10 ^*** 0.40±0.09 ^### Day 49 0.82±0.24 ^*** 0.74±0.18 ^*** 0.75±0.10 ^*** 0.43±0.10 ^##

Significant differences between the normal control group of group 1-1 and group 1-2 to group 1-8 were * p<0.05, ** p<0.01, *** p<0.001; Significant differences between the negative control group 1-2 and groups 1-3 to 1-8 were # p<0.05, ## p<0.01

As atopic dermatitis is continuously induced, the ear thickness level of all atopic-inducing groups except for groups 1-8 from day 28 (day 14 of test substance administration) to the end of the test from the initiation of atopic dermatitis is significantly compared to that of group 1-1. (P<0.001, p<0.01, or p<0.05), the ear thickness level of group 1-8 was significantly lower than that of the negative control group of group 1-2 (p<0.001 or p<0.01).

The group administered with thyme extract (TEWE) of group 1-3 and the group of thyme extract of group 1-6 and the juice mixture (TEmix2) had a tendency to slightly decrease ear thickness compared to the negative control group of group 1-2. And showed a similar tendency to that of the group 1-7 administration group.

4. Spleen weight measurement result

On the day of necropsy on the 49th day of the initiation of atopic dermatitis induction, the spleen was removed and the weight was measured, and the weight and the ratio to the body weight were shown in Tables 6 and 7.

division Group 1-1 Group 1-2 Group 1-3 Group 1-4 Weight(g) 29.98±2.29 27.67±1.23 25.90±2.40 27.44±1.85 Spleen weight (g) 0.0856±0.0086 0.1143±0.0300 0.1022±0.0358 0.1178±0.0460 Spleen weight to weight ratio (%) 0.2854±0.0182 0.4112±0.0941 0.3904±0.1231 0.4338±0.1828

division Group 1-5 Group 1-6 Group 1-7 Group 1-8 Weight(g) 26.00±1.84 27.42±1.34 26.33±2.46 24.19±1.32 Spleen weight (g) 0.0839±0.0221 0.0936±0.0241 0.0964±0.0230 0.0283±0.0028 ^***,### Spleen weight to weight ratio (%) 0.3224±0.0829 0.3413±0.0877 0.3666±0.0812 0.1169±0.0109 ^**,###

Significant differences between the normal control group of group 1-1 and group 1-2 to group 1-8 were * p<0.05, ** p<0.01, *** p<0.001; Significant differences between the negative control group 1-2 and groups 1-3 to 1-8 were # p<0.05, ## p<0.01

Compared to the normal control group of group 1-1, the absolute and relative spleen weights of the negative control group of group 1-2 were both increased, but among the test substance administration groups of group 1-3 to group 1-7, There was no statistically significant reduction in spleen weight compared to group 1-2.

However, in the case of the dexamethasone-administered group of groups 1-8, both absolute and relative weight levels of the spleen were significantly lower than those of groups 1-1 and 1-2 (p<0.001 and p<0.01), and In the case of case, it was found to be only about 25% of the group 1-2, and strong immunosuppression by dexamethasone. It was confirmed that the effect led to the contraction of the spleen.

5. Total leukocyte, neutrophil, eosinophil and IgE measurement results

Using the blood collected on the day of necropsy on the 49th day of the initiation of atopic dermatitis, the total white blood cell count was measured with an automatic cell count calculator (TC20 Automated Cell Counter, Biorad), and neutrophils and eosinophils were measured by flow cytometry. Was measured, and the IgE concentration in blood was measured by ELISA assay, and are shown in Tables 8 and 9.

division Group 1-1 Group 1-2 Group 1-3 Group 1-4 Whole white blood cells
(×10 ³ cells/µl) 1.75±0.34 11.03±5.06 ^*** 10.22±1.59 ^*** 5.43±1.82 ^## Neutrophil
(×10 ³ cells/µl) 0.02±0.01 0.39±0.28 ^*** 0.13±0.10 ^# 0.16±0.17 Eosinophil
(×10 ² cells/µl) 0.01±0.00 0.43±0.18 ^*** 0.10±0.04 ^### 0.11±0.11 ^### IgE
(㎍/mL) 0.1809±0.0709 11.0609±1.9891 ^*** 7.7147±1.8259 ^***,## 6.7752±2.3640 ^***,##

division Group 1-5 Group 1-6 Group 1-7 Group 1-8 Whole white blood cells
(×10 ³ cells/µl) 5.41±3.26 ^## 4.44±2.30 ^## 9.73±3.36 ^* 2.79±1.25 ^## Neutrophil
(×10 ³ cells/µl) 0.09±0.03 ^## 0.09±0.05 ^## 0.26±0.32 ^* 0.05±0.01 ^## Eosinophil
(×10 ² cells/µl) 0.07±0.04 ^### 0.07±0.02 ^### 0.25±0.25 ^**,# 0.04±0.01 ^### IgE
(㎍/mL) 5.1846±2.1722 ^***,### 6.4169±1.0060 ^***,### 9.5996.10±3.3080 ^*** 2.4256±0.7861 ^###

Significant differences between the normal control group of group 1-1 and group 1-2 to group 1-8 were * p<0.05, ** p<0.01, *** p<0.001; Significant differences between the negative control group 1-2 and groups 1-3 to 1-8 were # p<0.05, ## p<0.01

As a result of measuring the total white blood cell count, thyme extract administration group of group 1-4, thyme extract of group 1-5, and thyme extract of self-made cheonin-guk juice mixture (TEmix1) administration group, thyme extract of group 1-6 and thyme extract of self-made Cheonin-guk juice The total leukocyte level was significantly lower in the mixture (TEmix2) administration group and the dexamethasone administration group of groups 1-8 compared to the negative control group of group 1-2 (both p<0.01).

In the case of neutrophils, thyme extract administration group of group 1-3, thyme extract of group 1-5, and thyme extract from group 1-5 (TEmix1) administration group, thyme extract of group 1-6, and thyme extract from group 1-6 (TEmix2) The total leukocyte level was significantly lower in the dexamethasone administration group of the administration group and group 1-8 compared to the negative control group of group 1-2 (p<0.05, p<0.01, p<0.01, and p<0.01).

In addition, in the case of eosinophils, the negative control group 1-2 and the group 1-7 administration group of sativa extract were significantly higher than that of the positive control group 1-1 (p<0.001 and p<0.01). -Tyme extract administration group of groups 3 and 1-4, thyme extract of group 1-5, and thyme extract from group 1-5 (TEmix1) administration group, thyme extract from group 1-6, and thyme extract from group 1-6 (TEmix2) The administration group and the dexamethasone administration group of groups 1-8 were significantly lower than that of the negative control group of group 1-2 (both p<0.001).

Atopic dermatitis in the group administered with thyme extract of group 1-3, thyme extract of group 1-5, and thyme extract from group 1-5 (TEmix1) administration group, thyme extract of group 1-6 and thyme extract from group 1-6 (TEmix2) Significantly reduced both neutrophils and eosinophils, which play an important role in the pathologic mechanism of, in particular, was significantly better than the group administered with styrofoam extract of positive control group 1, group 1-7, and dexamethasone-treated group of positive control group 2, group 1-8. It was confirmed at a level similar to that of.

On the other hand, the blood IgE concentration was significantly higher in all atopy-inducing groups except for groups 1-8 than in the normal control group in group 1-1 (p<0.001), but in groups 1-3 and 1-4. The thyme extract administration group, the thyme extract of group 1-5, and the thyme extract of group 1-5 (TEmix1) administration group and the thyme extract of group 1-6 and the administration group of thyme extract (TEmix2) are negative control group of group 1-2 Was significantly lower than that of (p<0.01, p<0.01, p<0.001 and p<0.001). From the above results, thyme extract of group 1-3 and group 1-4, in particular, thyme extract of group 1-5 and thyme extract and extract of thyme from group 1-6 administration group (TEmix1) administration group and thyme extract of group 1-6 The liquid mixture (TEmix2) administration group was found to be more effective in inhibiting the production of IgE, which is a key factor in allergic reactions, than the extracts of the positive control group 1.

6. Results of measurement of inflammatory cell infiltration level and mast cell infiltration level in the epidermis

After the initiation of atopic dermatitis induction, the tissue collected on the day of necropsy on the 49th day was prepared for histopathological examination, and then Hematoxylin & Eosin (H&E) and toluidine blue staining were performed, and an optical microscope (Olympus BX53, Japan) was used to observe histopathological changes.

In the case of histopathological evaluation using Hematoxylin & Eosin (H&E) stained slides, the most severe lesion site was selected, and then the level of inflammatory cell infiltration in the epidermis was evaluated, and histopathological evaluation using toluidine blue stained slides. , After selecting the most severe lesion site, the number of mast cells was counted and shown in Tables 10 and 11.

division Group 1-1 Group 1-2 Group 1-3 Group 1-4 Infiltration of inflammatory cells in the epidermis (score) 0.0±0.0 2.9±0.4 ^*** 2.4±1.1 ^** 2.1±1.5 ^* Mast cells (dog) 20.5±4.2 95.4±11.5 ^*** 86.7±26.8 ^*** 82.1±17.7 ^***

division Group 1-5 Group 1-6 Group 1-7 Group 1-8 Infiltration of inflammatory cells in the epidermis (score) 0.9±1.5 ^# 1.3±1.6 3.0±0.0 ^*** 0.0±0.0 ^### Mast cells (dog) 70.9±17.1 ^***,# 70.3±18.4 ^***,# 64.8±18.2 ^***,## 41.2±12.4 ^###

Significant differences between the normal control group of group 1-1 and group 1-2 to group 1-8 were * p<0.05, ** p<0.01, *** p<0.001; Significant differences between the negative control group 1-2 and groups 1-3 to 1-8 were # p<0.05, ## p<0.01

In the negative control of group 1-2, the thyme extract-administered group of groups 1-3 and 1-4, and the thyme extract-administered group of group 1-7, the level of inflammatory cell infiltration in the epidermis was the normal control group of group 1-1. Was significantly higher than (p<0.001, p<0.01, p<0.05, and p<0.001).

Furthermore, the thyme extract of group 1-5 and the group administered with TEmix1 and dexamethasone of group 1-8 were significantly lower than that of the negative control group of group 1-2 (p<0.05 and p<0.001). ). In the case of groups 1-6, it was not significant, but the infiltration of inflammatory cells in the epidermis tended to decrease.

On the other hand, as a result of confirming the level of invasion of mast cells, the group 1-5 of thyme extract and Jasmine Cheonin-guk juice mixture (TEmix1) administration group, Group 1-6 of thyme extract and Jasmine Cheonin-guk juice mixture (TEmix2) administration group, The number of mast cells was significantly lower in the group 1-7 treated with the snail extract and the group 1-8 treated with dexamethasone compared to the negative control group in the 1-2 group (p<0.05, p<0.05, p<0.01, and p<0.001). ).

Experimental Example 2

1. Comparison of anti-inflammatory activity of extracts in mouse-derived macrophage RAW264.7

The anti-inflammatory activity of thyme ethanol extract (TVEE) of Preparation Example 2 and thyme ( Thymus quinquecostatus ) ethanol extract (TQEE) prepared using the method described in Preparation Example 2 were compared.

RAW264.7 cells, which are mouse-derived macrophages, were aliquoted into a 24-well plate at 4×10 ⁵ cells/well and cultured for 24 hours. After removing the culture medium, TVEE and TQEE diluted in serum-free medium were treated at a concentration of 250 μg/mL, respectively. At the same time, LPS (lipopolysaccharide) diluted in serum-free medium was treated at a final concentration of 500 ng/mL, and cultured for 24 hours. After incubation, the culture medium is centrifuged (5,000 rpm, 3 minutes, 4 ℃) to remove the suspended cells, and the supernatant is used to remove nitric oxide (NO), an inflammatory mediator secreted by LPS, and IL-, an inflammatory cytokine The amount of 1β and IL-6 secretion was measured.

Nitric oxide secretion was measured by mixing 50 μL Griess's reagent and 50 μL of supernatant in a 96-well plate, reacting at room temperature for 15 minutes, and then measuring the absorbance at 540 nm using a microplate reader to quantify the amount of nitric oxide secretion. The amount of secretion of kine IL-6 and IL-1β was analyzed using an ELISA kit (KOMA BIOTECH Co., Korea).

In addition, RBL-2H3 (rat mast cell line) cells, which are rat-derived mast cells, were purchased from the American Type Culture Collection (ATCC, USA) and cultured. In order to measure the amount of TNF-α secretion according to sample treatment in the RBL-2H3 cells, the cells were dispensed into a 24-well plate at 2×10 ⁵ cells/well and cultured for 24 hours. After removing the culture medium, samples diluted in serum-free medium were treated by concentration. At the same time, the mast cells were sensitized by treatment with A23187 1 μM and PMA 50 nM for 18 hours. After cultivation, the culture medium was centrifuged (5,000 rpm, 3 minutes, 4° C.), and the secreted TNF-α was quantified using an ELISA Kit (KOMA BIOTECH Co., Korea).

In the measurement of nitric oxide, IL-1β, and IL-6 secretion, group 2-1 was the normal control group that did not induce inflammation in cells with LPS, and the remaining 3 groups were treated with the test substance after inducing inflammation with LPS. Group 2-2 was a negative control treated with physiological saline, group 2-3 was treated with thyme ethanol extract (TVEE) 250 μg/mL, and group 2-4 was treated with thyme ethanol extract (TQEE) 250 μg/mL. . In the measurement of TNF-α secretion, group 2-1 was a normal control group that did not sensitize mast cells with PMA, and the other 3 groups were sensitized with PMA and treated with the test substance, and group 2-2 was treated with physiological saline. The negative control group, Group 2-3 were treated with thyme ethanol extract (TVEE) 250 μg/mL, and Group 2-4 were treated with thyme ethanol extract (TQEE) 250 μg/mL.

division Group 2-1 Group 2-2 Group 2-3 Group 2-4 Nitric acid (μM) -0.5±0.1 45.7±5.8 ^** 10.8±0 ^# 5.43±1.82 ^# IL-6 (pg/mL) 0±2 1571±111 ^* 66±11 ^## 116±12 ^## IL-1β (pg/mL) 10.8±0 46.5±1.2 ^* 2.1±0.3 ^## 9.4±1.4 ^## TNF-α (pg/mL) 5.6±1.7 71.3±7.8 ^* 36.8±7.0 ^# 48.6±6.3

Significant differences between the normal control group of group 2-1 and group 2-2 to group 2-4 were * p<0.05, ** p<0.01, *** p<0.001; Significant differences between the negative control of group 2-2 and groups 2-3 and 2-4 were # p<0.05, ## p<0.01

In the negative control group 2-2, which induced cell inflammation with LPS, the secretion of nitric oxide, IL-1β and IL-6 significantly increased compared to the normal control group 2-1 (p<0.01 or p<0.05). ). In addition, the negative control group of group 2-2, which sensitized mast cells with PMA, also significantly increased the amount of TNF-α secretion compared to the normal control group of group 2-1 (p<0.05).

The thyme ethanol extract (TVEE) treatment group of group 2-3 and the thyme ethanol extract (TQEE) treatment group of group 2-4 contained nitric oxide, IL-1β and IL-6 compared to the negative control group of 2-2. Significantly decreased (p<0.01 or p<0.05). Among them, the thyme ethanol extract (TVEE) treatment group of group 2-3 lowered the IL-1β and IL-6 secretion significantly increased by LPS treatment than the thyme ethanol extract (TQEE) treatment group of group 2-4. Showed a trend.

In addition, thyme ethanol extract (TVEE) treatment group of group 2-3 significantly lowered the amount of TNF-α secretion significantly increased by PMA sensitization in the negative control group 2-2 (p<0.05), but 2-4 Thyme ethanol extract (TQEE) treatment group of the group did not have a significant difference in the amount of TNF-α secretion from the negative control group of 2-2.

Therefore, thyme ethanol extract (TVEE) has an excellent anti-inflammatory effect of lowering the secretion of inflammatory cytokines IL-6, IL-1β and TNF-α compared to thyme ethanol extract (TQEE), so it is more advantageous in improving allergic or inflammatory skin diseases. It was judged as the material.

Experimental Example 3

The ratio of the total leukocytes of eosinophils, which are highly related to atopic dermatitis, and the absolute amount of thyme extract alone and a mixture of thyme extract and Jasper Cheonin Guk juice were measured and compared.

Based on the solid content of the Jagalcheoninguk juice extract (EPJ) and the thyme ethanol extract (TVEE) of Preparation Example 2 of Preparation Example 3, respectively, 9:1, 7:3, 5:5, 3 : 7 and 1: 9 were mixed in a weight ratio to prepare a powder mixture of Jajucheoninguk juice and thyme extract.

Experimental animals, breeding conditions and methods of inducing atopic dermatitis were all carried out in the same manner as in Experimental Example 1, and the normal and negative controls were also used in the same manner as in Experimental Example 1, and all samples were prepared using solids of extracts, juices, or mixtures thereof. As a standard, 200 mg/kg was orally administered once a day for 4 weeks based on a 10 mL/kg dose. The normal control group and the negative control group were orally administered 10 mL/kg of physiological saline as a vehicle.

The percentage of leukocytes was confirmed using the blood collected on the day 42 of necropsy on the 42nd day of induction of atopic dermatitis. Some of the collected blood was injected into the CBC bottle containing the anticoagulant EDTA-2K, and then the ratio and absolute amount of eosinophils were checked using an automatic blood analyzer (ADVIA 2120, SIEMENS, USA), and are shown in Table 13.

division Explanation Eosinophil (%) Eosinophil (×10 ³ cell/µl) Group 3-1 Normal control 2.0±0.6 0.05±0.03 Group 3-2 Negative control 5.0±3.8 ^** 0.09±0.06 ^* Group 3-3 Time alone 1.9±0.5 ^## 0.03±0.01 ^### Group 3-4 Independence 1.7±0.6 ^### 0.04±0.02 ^## Group 3-5 9:1 (independent heavenly country: time) 1.5±0.8 ^### 0.02±0.8 ^### Group 3-6 7:3 (independent heavenly country: time) 2.6±0.9 ^# 0.05±0.01 ^# Group 3-7 5:5 (independent heavenly country: time) 2.2±0.8 ^## 0.04±0.02 ^## Group 3-8 3:7 (independent heavenly people: time) 1.8±1.0 ^### 0.04±0.02 ^## Group 3-9 1:9 (independent heavenly people: time) 1.9±1.4 ^## 0.03±0.02 ^###

Significant differences between the normal control group of group 3-1 and groups 3-2 to 3-9 were * p<0.05, ** p<0.01; Significant differences between the negative control group 3-2 and groups 3-3 to 3-9 were # p<0.05, ## p<0.01, ### p<0.001

In atopic dermatitis, the number of inflammation-related cells, in particular, the ratio of eosinophils is increased, and eosinophils are known to play an important role in the pathological mechanisms of atopic dermatitis.

The proportion of eosinophils and the absolute amount of eosinophils significantly and significantly increased in the negative control group of group 3-2 compared to the normal control group of group 3-1 (p<0.01 and p<0.05), and group 3-3 to group 3 All test substance administration groups of groups 3-9 significantly decreased compared to the negative control group of group 3-2 (p<0.05, p<0.01, and p<0.001).

Among the test groups in which the thyme ethanol extract (TVEE) was mixed with the extract (EPJ), the group 3-5 was mixed in a ratio of 9: 1 to the thyme alone and the group 3- It was confirmed that the proportion of eosinophils and the absolute amount of eosinophils were significantly reduced compared to the group 4 administered with independent Cheonin Guk.

Therefore, for the treatment of atopic dermatitis, thyme extract alone is also effective, but preferably thyme extract and Jasper Cheonin Guk juice are used in a solid content of 1: 99 in a weight ratio of 20: 80, preferably in a weight ratio of 5: 95 to 15: 85. It is believed that mixing by weight ratio may be advantageous.

Statistical analysis

The experimental results are expressed as mean ± standard deviation, and in the case of parametric multiple comparisons, the normality of the data was assumed, and the results were tested by one-way ANOVA, and if the results were significant, Dunnett's multiple comparison test Post-test was performed using, and significant differences between the test groups were analyzed. In the case of nonparametric multiple comparisons, the test was performed with the Kruskal-Wallis'H-test, and if the result was significant, the significant difference between the test groups was analyzed using the post-analysis Mann-Whitney U test.

Statistical analysis was performed using Prism 7.04 (GraphPad Software Inc., San Diego, CA, USA), and a p value of less than 0.05 was determined to be statistically significant.

Hereinafter, an example of a formulation of a composition comprising a thyme extract of the present invention, or a mixture of thyme extract and Jajucheonin-guk juice, is described, but the present invention is not intended to limit it, but is intended to be described in detail.

Formulation Example 1: Preparation of powder

20 mg of thyme extract (TVWE) powder of Preparation Example 1

100 mg lactose

10 mg of talc

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

Formulation Example 2: Preparation of tablets

Preparation Example 2 thyme extract (TVEE) powder 10 mg

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

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

Formulation Example 3: Preparation of Capsule

Preparation Example 4 thyme extract and Jajucheoninguk juice powder 10 mg

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

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

Formulation Example 4: Preparation of granules

Thyme extract of Preparation Example 5 and 1,000 mg of Jasper Cheonin Guk juice powder

Vitamin mixture right amount

Vitamin A acetate 70 ㎍

1.0 mg of vitamin E

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

1.7 mg of nicotinic acid amide

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

1.75 mg ferrous sulfate

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

90 mg of potassium citrate

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

As for the composition ratio of the vitamin and mineral mixture, ingredients suitable for health functional foods are mixed in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a general health functional food manufacturing method. Next, granules are prepared and can be used to prepare a health functional food composition according to a conventional method.

Formulation Example 5: Preparation of beverage formulation

1,000 mg of thyme extract (TVWE) powder of Preparation Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

Total 900 mL with purified water

After mixing the above ingredients according to a conventional beverage manufacturing method, after stirring and heating at 85°C for about 1 hour, the resulting solution was filtered and obtained in a sterilized 2 L container, and sealed and sterilized.

Formulation Example 6: Preparation of feed composition

The thyme extract of Preparation Example 4 and the extract (TEmix1) powder 0.1 kg, corn 25.5 kg, wheat 15.04 kg, wheat flour 8.15 kg, rice bran 7.4 kg, soybean meal 18 kg, octane 1 kg, chicken by-products 14 kg, animal origin Oil 9 kg, processed salt 0.3 kg, tricalcium phosphate 0.3 kg, limestone 1 kg, choline chloride 0.01 kg, vitamins 0.05 kg, minerals 0.05 kg and digestive enzymes 0.1 kg were mixed to prepare an animal (dog, dog) feed composition.

Claims (15)

Pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases comprising thyme extract as an active ingredient. The method of claim 1, wherein the allergic or inflammatory skin disease is skin inflammation, eczema, contact dermatitis, atopic dermatitis, seborrheic dermatitis, chronic lichen planus, interstitial dermatitis, deprived dermatitis, papular urticaria, psoriasis, psoriatic arthritis, sun dermatitis. , A pharmaceutical composition for treating or preventing allergic or inflammatory skin diseases, characterized in that any one skin disease selected from sunburn and acne. The pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases according to claim 1, wherein the thyme is a stem, leaf, or above-ground outpost of thyme. The pharmaceutical composition for treating or preventing allergic or inflammatory skin diseases according to claim 1, wherein the extract is an extract using water, alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The pharmaceutical composition for the treatment or prevention of an allergic or inflammatory skin disease according to claim 1, further comprising a juice of Jagal Cheonin Guk. [6] The pharmaceutical composition for treating or preventing allergic or inflammatory skin diseases according to claim 5, wherein the mixing ratio of thyme extract and Japonica Cheonin Guk juice is a solid content-based mixing ratio of 1:99 to 99:1 by weight. The pharmaceutical composition for treating or preventing allergic or inflammatory skin diseases according to claim 1, wherein the pharmaceutical composition is an oral preparation. Animal pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases comprising thyme extract as an active ingredient. [9] The veterinary pharmaceutical composition for the treatment or prevention of allergic or inflammatory skin diseases according to claim 8, wherein the pharmaceutical composition further comprises a juice of Jagal Cheonin Guk. Food composition for improving allergic or inflammatory skin diseases comprising thyme extract as an active ingredient. 11. The food composition for improving allergic or inflammatory skin diseases according to claim 10, wherein the food composition further comprises a juice of Jajucheonin-guk. The food composition for improving allergic or inflammatory skin diseases according to claim 10, wherein the food composition is a powder, granule, tablet, capsule, pill, extract, jelly formulation, tea bag formulation, or beverage formulation. The food composition according to claim 10, wherein the food composition is a health functional food for improving allergic or inflammatory skin diseases. Feed composition for improving allergic or inflammatory skin diseases comprising thyme extract as an active ingredient. [15] The feed composition for improving allergic or inflammatory skin diseases according to claim 14, wherein the feed composition further comprises a juice of Jagalcheonin-guk.