KR20180020195A - Pharmaceutical composition comprising Cymbidium extracts for preventing or treating allergic diseases or contact - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating allergic diseases or contact dermatitis, comprising a cymbidium extract as an effective component, an external application for skin, a cosmetic composition, and a functional health food. In particular, the cymbidium extract shows the effect of inhibiting the secretion of mast cells causing an allergy in a low concentration, and shows the effect of reducing scratching behavior, skin sensation, and IgE in blood in an animal model of allergic contact dermatitis caused by antibodies of delayed-type hypersensitivity reactions (DNCB (2,4-dinitrochlorobenzene)), and thus may be beneficially used in a pharmaceutical composition for preventing and treating allergic diseases or contact dermatitis, an external application for skin, a cosmetic composition, and a functional health food.

Description

The present invention relates to a pharmaceutical composition for preventing or treating allergic diseases or contact dermatitis comprising cymbidium extract as an active ingredient,

The present invention relates to a pharmaceutical composition for preventing or treating an allergic disease or contact dermatitis containing Cymbidium extract as an active ingredient, a health functional food for preventing or improving allergic diseases or contact dermatitis, an external preparation for skin and a cosmetic composition .

Mast cells and blood neutrophils secrete cytokines, which cause various allergic diseases such as allergic rhinitis, atopic dermatitis, allergic dermatitis, allergic conjunctivitis, allergic asthma, food allergy and anaphylactic shock (Galli SJ et al., Agents & Actions supplements., 1993). These cells have a receptor for IgE (FcεRI), an antibody that induces an allergen on the cell surface, which is stimulated by allergenic substances (called antigens or allergens) and causes a variety of allergies The substance is secreted out of the cell (Amin K., Respiratory Medicine., 2012).

There are many ways to treat allergies, but most of the current allergic therapies are being studied to alleviate symptoms rather than to eliminate them. In general, the major market is antagonists to receptors such as histamine and leukotrienes secreted by allergens in mast cells, and these drugs have become a huge market (A Roquet et al., American Journal of Respiratory and Critical Care Medicine, 1997). However, these drugs show resistance to the patient within a short period of time after administration, so that they often fail to improve their symptoms after a certain period or after repeated administration. Therefore, it is demanding the development of anti-allergic drugs without side effects such as antihistamines.

In addition to this, there is a method of identifying allergens for allergy sufferers suffering from allergies and administering them in small doses for several years to gradually reduce the allergies. However, this method has several disadvantages that it takes several years to treat and can cause anaphylactic shock. Methods of using other DNA vaccines (Jay E. Slater et al., Journal of Allergy and Clinical Immunology., 1998), treatment to block IgE binding to receptors in mast cells (Brian J Sutton et al., British Medical Bulletin., 2000), and therapeutic approaches to IL-4, an allergen-inducing cytokine (Carl J., The Journal of Experimental Medicine., 1993). However, these approaches are costly or have yet to be fully addressed.

Cymbidium , which is used as an active ingredient in the composition of the present invention, is originated from Southeast Asia, China, Japan, New Guinea and Australia, and is composed of two petals, one petal, one petal, three calyxes, It has roots like a spindle-like bloom and bulky string, commonly called a bulb. It has been reported that it has more negative ion generation and higher relative humidity than Siberiania, so it has excellent air purification and humidification ability. In addition, it has been developed and sold as a hair regeneration product, hair shampoo, etc. in Japan, and the results of these studies have been reported in Korea. According to a number of studies, phenanthrene and phenylpropanoid extracted from the roots of Cymbidium have antimicrobial activity against Bacillus, Pneumoniae, and Red Pheceae in the antibacterial test (Yoshikawa K et al., Journal of Natural Medicines., 2014) and carrageenin (Howlader MA et al., Pakistan Journal of Biological Sciences., 2011), flavoring and cosmetic ingredients Linalool and 4-methyl-phenol, which are known to be used as liposomes, have been analyzed by chromatographic methods in Cymbidium roots (Gaytan VG et al., Journal of Analytical Sciences, Methods and Instrumentation. 2013). In addition, aromatic glucosides, an active ingredient extracted from cymbidium, have been shown to exhibit antioxidant activity in peroxide removal experiments (Yoshikawa L et al., Journal of Natural Medicines., 2013).

Korean Patent Laid-Open Publication No. 2013-0010885 discloses that Dionaea muscipula extract may be included in a composition for cosmetic treatment against skin changes due to accelerated or endogenous aging and further comprising Cymbidium erythraeum ) extract may be included to combat the loss of wrinkles, firmness and elasticity and increased coloration.

However, anti-allergic or contact dermatitis-inhibiting effects on cymbidium are not known.

Korean Patent Publication No. 2013-0010885

Yoshikawa K et al., Journal of Natural Medicines., 2014 Howlader MA et al., Pakistan Journal of Biological Sciences., 2011 Gaytan V G et al., Journal of Analytical Sciences, Methods and Instrumentation., 2013 Yoshikawa L et al., Journal of Natural Medicines., 2013

It is an object of the present invention to find new materials for the prevention, treatment or amelioration of allergic diseases or contact dermatitis containing cymbidium extract as an active ingredient and to provide new uses thereof.

To achieve the above object, the present invention is by identifying the Cymbidium (cymbidium) secretion of the allergens of mast cells to cause allergy due to extract inhibitory effect and inflammation inhibition in contact dermatitis in animal models effect the Cymbidium (cymbidium The present invention provides a pharmaceutical composition for preventing or treating an allergic disease or contact dermatitis containing an extract as an active ingredient, a health functional food for preventing or improving allergic diseases or contact dermatitis, an external preparation for skin and a cosmetic composition.

The cymbidium extract of the present invention is effective for inhibiting the secretory action of allergen-inducing substances of mast cells inducing allergy and exhibiting delayed hypersensitivity (DNCB (2.4-dinitrochlorobenzene)) without causing cytotoxicity at therapeutic doses The cymbidium extract is useful as a pharmaceutical composition for the prevention or treatment of allergic diseases or contact dermatitis because it exhibits effects such as reduction of scratching activity and evaluation of skin sensation in an animal model of contact dermatitis, , An allergic disease or a health functional food for preventing or improving contact dermatitis, an external preparation for skin, and a cosmetic composition.

FIG. 1 is a graph showing the cytotoxicity observed when cymbidium extract was cultured in mast cells after treatment with 25, 50, 100 and 200 μg / ml.
FIG. 2 shows the effect of inhibiting the secretion of allergen-inducing substances in mast cells according to the concentration of cymbidium extract.
FIG. 3 is a photograph and a graph showing concentration-dependency of the inhibitory effect of cymbidium extract on in vivo antibody / antigen response.
Fig. 4 is a graph showing the scratching behavior of ICR mice when cymbidium extract was applied.
FIG. 5 is a graph showing skin sensory evaluation of ICR mice when cymbidium extract was applied.
FIG. 6 is a graph showing blood IgE levels of ICR mice when Cymbidium extract was applied.

The present invention relates to a pharmaceutical composition for preventing or treating allergic diseases or contact dermatitis containing cymbidium extract as an active ingredient.

In one embodiment of the present invention, the cymbidium extract may be any one selected from the group consisting of flowers, leaves, branches, roots, stem, fruit, skin and mixtures thereof of Cymbidium, and more specifically, Roots and stems can be used.

In one embodiment of the present invention, the cymbidium extract is extracted with water, an organic solvent, or a mixed solvent thereof.

In an embodiment of the present invention, the organic solvent may be an alcohol having 1 to 5 carbon atoms, ethyl acetate, acetone, ether, chloroform, benzene, hexane or dichloromethane, more specifically ethanol.

In one embodiment of the present invention, the allergic disease may be a disease selected from the group consisting of atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, allergic asthma and anaphylactic shock, It is not limited thereto.

 In one aspect of the invention, the allergic dermatitis can be psoriasis, contact allergic dermatitis, urticaria, and the like.

 In one aspect of the invention, the pharmaceutical composition may alleviate or inhibit inflammatory reaction, itching, or edema due to allergic diseases or contact dermatitis.

In one embodiment of the present invention, the contact dermatitis is contact dermatitis due to contact with an external substance.

In one aspect of the present invention, the cymbidium in the pharmaceutical composition may be contained in an amount of 0.001 to 10% by weight of the total pharmaceutical composition.

In one aspect of the present invention, the pharmaceutical composition may be formulated into a formulation selected from the group consisting of tablets, granules, pills, capsules, solutions, injections, ointments, suppositories and powders, but is not limited thereto.

The pharmaceutical composition for the prevention and treatment of allergic diseases or contact dermatitis containing cymbidium extract according to the present invention may be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, , External preparations (ointments, patches, gels), suppositories, and parenteral sterile injectable solutions.

Examples of carriers, excipients and diluents that can be contained in the composition containing cymbidium extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, aceitol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylidrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, for example starch, calcium carbonate, sucrose, Or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

The composition of the present invention is administered in a pharmaceutically effective amount.

The term "pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will vary depending on the species and severity, age, sex, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. However, for the desired effect, the cymbidium extract of the present invention may be administered in an amount of 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg once or several times a day for an adult. The dose is not intended to limit the scope of the invention in any way.

The composition of the present invention may be administered in the form of an individual therapeutic agent or in combination with other therapeutic agents exhibiting antiallergic or contact dermatitis-inhibiting effects, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

The term "individual" as used herein refers to all animals, including humans, who have already developed or are capable of developing a disease that can be prevented or treated through the use of antiallergic or contact dermatitis inhibiting activity, and includes a composition comprising the extract of the present invention By administering to an individual, the disease can be effectively prevented and treated.

The route of administration of the composition may be administered via any conventional route so long as it can reach the target tissue. The composition of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermally, orally, intranasally, intrapulmonarily, or rectally, though it is not intended to be limited thereto. The composition may also be administered by any device capable of transferring the active agent to the target cell.

In the present invention, "treatment" includes the partial cure, improvement and alleviation of allergic diseases or contact dermatitis symptoms as well as curing of allergic diseases or contact dermatitis as a result of applying the pharmaceutical composition of the present invention to allergic diseases or contact dermatitis do.

In the present invention, "prevention" means that the pharmaceutical composition of the present invention is applied to allergic diseases or contact dermatitis to prevent or prevent allergic diseases or contact dermatitis symptoms so that allergy diseases or contact dermatitis are prevented from occurring in advance it means.

In the present invention, "improvement" is meant to include alleviation, prevention or treatment of symptoms.

In the present invention, the term "active ingredient" means an ingredient exhibiting activity alone or together with a carrier which is not active per se.

The present invention relates to an external preparation for skin for preventing or treating allergic diseases or contact dermatitis containing cymbidium extract as an active ingredient.

In one embodiment of the present invention, the cymbidium extract may be any one selected from the group consisting of flowers, leaves, branches, roots, stem, fruit, skin and mixtures thereof of Cymbidium, and more specifically, Roots and stems can be used

In one embodiment of the present invention, the cymbidium extract is extracted with water, an organic solvent, or a mixed solvent thereof.

In one embodiment of the present invention, the organic solvent may be an alcohol having 1 to 5 carbon atoms, ethyl acetate, acetone, ether, chloroform, benzene, hexane or dichloromethane, and more specifically, ethanol.

In one embodiment of the present invention, the allergic disease may be a disease selected from the group consisting of atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, allergic asthma and anaphylactic shock, It is not limited thereto.

In one aspect of the invention, the allergic dermatitis may be psoriasis, contact allergic dermatitis, urticaria, and the like.

In one embodiment of the present invention, the external preparation for skin may alleviate or suppress inflammatory reaction, itching, or swelling caused by allergic diseases or contact dermatitis.

In one aspect of the present invention, the external preparation for skin may be formulated into a formulation selected from the group consisting of external preparation, external preparation, external preparation, ointment, cream, gel, warning, dressing, patch, It is not limited thereto.

In one embodiment of the present invention, the cymbidium in the external preparation for skin may be 0.0001 to 5% by weight of the total external preparation for skin.

The present invention relates to a health functional food for preventing or ameliorating allergic diseases or contact dermatitis containing cymbidium extract as an active ingredient.

In one embodiment of the present invention, the cymbidium extract may be any one selected from the group consisting of flowers, leaves, branches, roots, stem, fruit, skin and mixtures thereof of Cymbidium, and more specifically, Roots and stems can be used.

In one embodiment of the present invention, the cymbidium extract is extracted with water, an organic solvent, or a mixed solvent thereof.

In one embodiment of the present invention, the organic solvent may be an alcohol having 1 to 5 carbon atoms, ethyl acetate, acetone, ether, chloroform, benzene, hexane or dichloromethane, and more specifically, ethanol.

In one embodiment of the present invention, the allergic disease may be a disease selected from the group consisting of atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, allergic asthma and anaphylactic shock, It is not limited thereto.

In one aspect of the invention, the allergic dermatitis may be psoriasis, contact allergic dermatitis, urticaria, and the like.

In one aspect of the present invention, the health functional food may alleviate or suppress inflammatory reaction, itching, or swelling caused by allergic diseases or contact dermatitis.

In one embodiment of the present invention, cymbidium in the health functional food may be contained in an amount of 0.001 to 1% by weight of the total health functional food.

In one aspect of the present invention, the health functional food may be formulated into a formulation selected from the group consisting of tablets, granules, pills, capsules, liquids and powders, but is not limited thereto.

The health functional food of the present invention may include an appropriate food supplementary additive including the above cimbury extract or a pharmaceutically acceptable salt thereof.

The term "food supplementary additive " in the present invention means a component which can be added to foods in a supplementary manner, and it can be appropriately selected and used by those skilled in the art as added to produce health functional foods of each formulation. Examples of food-aid additives include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, , a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated drink. However, the types of the food auxiliary additives of the present invention are not limited by these examples.

The term "health functional food" in the present invention refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and circles by using raw materials and components having useful functions in the human body. The term "functional" as used herein means that the structure and function of the human body have a beneficial effect on health uses such as controlling nutrients or physiological actions. The health functional food according to the present invention can be manufactured by a method commonly used in the art and can be prepared by adding raw materials and components that are conventionally added in the art. In addition, unlike general medicines, there is an advantage that there is no side effect that may be caused when a medicine is used for a long time by using a food as a raw material, and it is excellent in portability and the health functional food of the present invention is effective for improving anti- It can be taken as an adjuvant for.

The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the cymbidium extract of the present invention may be contained in an amount of 0.01 to 5% by weight in the raw material composition when the food is prepared. In the case of health drinks, it may be added at a rate of 0.02 to 2 g or 0.3 to 1 g based on 100 mL. However, in the case of long-term ingestion intended for health and hygiene purposes or for the purpose of controlling health, the amount can also be used in the above-mentioned range.

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient as well as ordinary foods. The above-mentioned natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.1 to 10 g, specifically about 1 to 8 g, more specifically about 5 to 6 g per 100 mL of the composition of the present invention.

In addition to the above, the cymbidium extract of the present invention may contain various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, coloring agents and aging agents (cheese, chocolate etc.), pectic acid and its salts, alginic acid and its salts, Colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.

In addition, the cymbidium extract of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. At this time, although the proportion of the additive is not critical, it is generally selected in the range of 0.0001 to about 10 parts by weight per 100 parts by weight of the cymbidium extract of the present invention.

The present invention relates to a cosmetic composition for preventing or improving an allergic disease or contact dermatitis containing cymbidium extract as an active ingredient.

In one embodiment of the present invention, the cymbidium extract may be any one selected from the group consisting of flowers, leaves, branches, roots, stem, fruit, skin and mixtures thereof of Cymbidium, and more specifically, Roots and stems can be used

In one embodiment of the present invention, the cymbidium extract is extracted with water, an organic solvent, or a mixed solvent thereof.

In one embodiment of the present invention, the organic solvent may be an alcohol having 1 to 5 carbon atoms, ethyl acetate, acetone, ether, chloroform, benzene, hexane or dichloromethane, and more specifically, ethanol.

In one embodiment of the present invention, the allergic disease may be a disease selected from the group consisting of atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, allergic asthma and anaphylactic shock, It is not limited thereto.

In one aspect of the invention, the allergic dermatitis may be psoriasis, contact allergic dermatitis, urticaria, and the like.

In one aspect of the present invention, the cosmetic composition may alleviate or inhibit inflammatory reaction, itching, or swelling caused by allergic diseases or contact dermatitis.

In one embodiment of the present invention, the cymbidium in the cosmetic composition may be 0.000001 to 1% by weight, more specifically 0.000001 to 0.01% by weight, more specifically 0.00005 to 0.001% by weight, of the total weight of the cosmetic composition.

In one aspect of the present invention, the cosmetic composition is a cosmetic composition of skin-adhesive type, for example, a base product cosmetic (lotion, cream, essence, cleansing foam, cleansing water, pack, soap) (Lipstick, mascara, makeup base), hair cosmetics (shampoo, rinse, hair conditioner, hair gel), and the like. In addition, it may be manufactured through a transdermal dosage form, for example, an ointment, a liquid, a dressing, a patch or a spray, but is not limited thereto.

In one embodiment of the present invention, the cosmetic composition may be used in an amount of about 0.05 to 10.0% by weight based on the dry weight of the cosmetic.

In one aspect of the present invention, the cosmetic composition may further comprise at least one of a fatty substance, an organic solvent, a solubilizing agent, a thickening and gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizing agent, a foaming agent, , Ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And may contain adjuvants commonly used in the cosmetics field, such as any of the other ingredients.

Hereinafter, the present invention will be described in more detail by way of Examples, Experimental Examples and Preparation Examples according to the present invention, but the scope of the present invention is not limited by the following Examples.

[ Example ]

Example  One. Cymbidium  Preparation of extract

Cymbidium was cultivated by the RDA's Good Agricultural Practice (GAP), and in 2009 Chungcheongbuk-do Voice (GPS: E 128 ° 62 'N 36 ° 56') It was harvested. Extraction of the sample was performed as follows. 50 g of Cymbidium root and stem dry samples were added with 150 ml of 99% ethanol, and the mixture was reacted at 150 rpm for 48 hours. After filtration, the powder was concentrated under reduced pressure and used as a sample of this experiment.

[ Experimental Example ] Physiological activity confirmation experiment

Experimental Example  One: Cymbidium  Confirmation of allergy inhibitory effect of extract

(1) Cell culture

Normal mast cells (RBL-2H3) were cultured in DMEM medium containing 5% fetal bovine serum (FBS). Normal mast cells were cultured in 75 ㎠ plastic flask (Falcon Co., England) with 10% FBS, 7.5% NaHCO ₃ 150 μg / ml, glutamine 58.4 μg / ml and antibiotics / antimycotics 4.4 μl / ml Were cultured in DMEM medium containing 5% CO ₂ at 37 ° C. Cells were maintained in secondary culture once every 2-3 days.

(2) Cell quantification

The medium was removed from the 75-cm plastic flask in which the cells were grown, washed with CMF-PBS (calcium magnesium free-phosphate buffered saline, pH 7.2), treated with 0.25% trypsin / EDTA and the cells were removed from the bottom of the flask The cells were neutralized with the culture medium and centrifuged (1200 rpm, 3 min). The culture medium was added to the pellet of the remaining cells and repeatedly inhaled with a sterile pipette to make a single cell suspension. Trypan blue was mixed with the cell suspension at a ratio of 9: 1, were measured using a hemocytometer.

(3) Cytotoxicity measurement

To determine the toxicity of cymbidium extract, MTT assay was used. That is, normal mast cells (RBL-2H3) were dispensed in a 24-well plate at 1 × 10 ⁵ cells / well. The cells were cultured in an incubator at 37 ° C and 5% CO ₂ for 24 hours and cultured for 24 hours at a concentration of 0, 25, 50, 100 and 200 μg / ml. MTT solution obtained by dissolving 0.01 g of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) in 10 ml of CMF-PBS (calcium magnesium free-phosphate buffered saline, pH 7.2) 50 [mu] l of each well was dispensed, and after the reaction was performed in an incubator for 1 hour, the medium was removed. Then, DMSO (dimethyl sulfoxide) was dispensed and transferred to 96 wells, respectively, and the absorbance was measured to determine the cytotoxicity of the cymbidium extract.

The cytotoxicity of cultured normal mast cells containing cymbidium extract is shown in FIG. The survival rate was 100% when the cymbidium extract was not added and the survival rate was similar to that when the cymbidium extract was added. Therefore, it was found that cymbidium extract did not show toxicity to cultured obese cells.

(4) Cymbidium  Inhibitory effect of extracts on mast cell-induced secretion of allergen

To determine whether cymbidium extract inhibits the secretion of allergens from mast cells inducing allergy in the body, mast cells were cultured in a 24-well plate at 2 × 10 ⁵ cells with 400 ng / ml DNP-specific IgE Lt; / RTI > The cultured cells were washed with PIPES buffer (25 mM PIPES, pH 7.2, 119 mM NaCl, 5 mM KCl, 0.4 mM MgCl ₂ , 1 mM CaCl ₂ , 5.6 mM glucose, and 0.1% BSA) Pre-incubated for 30 min. After preincubation, the antigen was added to a final concentration of 50 ng / ml to induce stimulation. The degree of secretion of the allergen inducer was determined by measuring the activity of hexosaminidase, a marker of degranulation secreted in the medium, from p-nitrophenylacetyl-β-D-glucosaminide The amount of p-nitrophenol liberated was determined.

FIG. 2 shows a concentration-dependent inhibitory effect of cymbidium extract on allergen-inducing mast cells. It was confirmed that the cymbidium extract inhibited the secretion of various allergens in mast cells in a concentration-dependent manner.

(5) Antibody / antigens inhibitory effect of cymbidium extract on passive skin anaphylaxis

In order to confirm the in vivo antibody / antigen inhibition effect of cymbidium extract, 0.5 μg of DNP-specific IgE antibody was injected into one ear of 7-week-old male ICR mice, and after 24 hours, 200 μg of DNP-BSA and Evans blue) 3% were injected intravenously via mouse tail vein to induce antigen / antibody reaction. After 1 hour, cymbidium extract was orally administered at the concentrations of 0, 30, 50, and 100 mg / kg. After 1 hour, ICR mice were euthanized and ears were injected with IgE antibody. The ear of the stained ICR mouse was treated with 500 μl of formamide and allowed to react for 12 hours at 63 ° C.

FIG. 3 shows concentration-dependent inhibitory effects of cymbidium extract on in vivo antibody / antigen response, and it was confirmed that cymbidium extract inhibited antibody / antigen response in ICR mice in a concentration-dependent manner.

Experimental Example 2: Inhibitory Effect of Cymbidium Extract on Atopic Dermatitis Induced Animal Model

(1) preparing an allergic contact dermatitis-inducing animal model

To induce allergic contact dermatitis, 7 weeks old male ICR mice were shaved at the dorsal hairs and 24 hours later, DNCB (2,4-dinitrochlorobenzene) was added to the mixture as a mixture of olive oil and acetone (Mixed volume ratio 1: 3) to 1 w / v%, and then 300 μl was applied to the back of the ICR mouse 2 cm ² for primary immunization. After 4 days of primary immunization, 250 μl of 0.5% DNCB was applied to the dorsal area, and the same immunosuppression was continued for about 2 weeks. In the group to which cymbidium extract was applied, 200 μL of dumbbell was applied at a dose of 10 mg / mL once every other day from the second DNCB administration.

(2) Measurement of scraping behavior change of allergic contact dermatitis-induced animal models

After the 2nd DNCB injection, video recording was performed at intervals of 48 hours, and a scratching behavior test was performed to measure the number of scratches caused by contact dermatitis. After the second application of DNCB, damage to the dorsal epidermis and persistent inflammation were observed, and the scratching behavior of the dorsal area of the individuals continued to increase.

FIG. 4 shows the scratching behavior of the ICR mouse when the cymbidium extract was applied. In the group to which the cymbidium extract was applied, the scratching behavior was significantly reduced after the fourth administration.

(3) Skin sensory evaluation of allergic contact dermatitis-induced animal models

Secondary Dose of the ICR mouse during the scratching behavior of DNCB after the administration of DNCB, itching and dry skin, edema and hematoma, wrinkles, folliculitis, no symptoms for these 5 items 0 point, symptom weakness 1 point, normal 2 Point score, and three points of severity. Then, the score of 5 items was added to the score between the minimum score of 0 (no symptoms) and the maximum score of 15 (all symptoms were severe).

FIG. 5 shows skin sensory evaluation of ICR mice when cymbidium extract was applied. When the skin sensory evaluation was performed to compare the DNCB-coated group with the Cymbidium extract-treated DNCB group, the DNCB- The score increased to 12 points as the number of cymbidium extracts increased, but it decreased from the third dose to 2 points.

(4) Serum of allergic contact dermatitis-induced animal models IgE Change of

 To measure serum IgE levels in allergic contact dermatitis induced animal models, DNCB and cymbidium extracts were performed up to six times, and cardiac blood was collected to separate the serum. Thereafter, the absorbance was measured at 450 nm using IgE enzyme-linked immunosorbent assay (ELISA MAXTMDeluxeset) for mouse.

FIG. 6 shows the serum IgE level of ICR mice when cimbuemia extract was applied. In the group that induced contact dermatitis only with DNCB, the concentration of IgE in serum was increased about 3 times. However, In the control group.

[ Manufacturing example ]

Manufacturing example  One. Sanje  Produce

Cymbidium extract 10 mg

Sucrose 100 mg

Talc 10 mg

The above components are powdered and mixed, and filled in an airtight container to prepare a powder.

Manufacturing example  2. Preparation of tablets

Cymbidium extract 10 mg

Starch 100 mg

Sucrose 100 mg

Magnesium stearate 2 mg

The tablets are prepared by mixing the above components according to a conventional method for producing tablets and then tableting them.

Manufacturing example  3. Preparation of capsules

Cymbidium extract 10 mg

Crystalline cellulose 3 mg

Lactose 15 mg

1 mg of magnesium stearate

The above components are mixed according to a conventional method for preparing a capsule, and then filled in a gelatin capsule to prepare a capsule.

Manufacturing example  4. Preparation of granules

Cymbidium extract 10 mg

Soybean extract 50 mg

Glucose 200 mg

Starch 500 mg

After mixing the above components, 100 mL of 30% ethanol is added, and the mixture is dried at 60 ° C to form granules, which are filled in a capsule to prepare granules.

Manufacturing example  5. Pennant  Produce

Cymbidium extract 20 mg

Lactose 1,500 mg

Glycerin 1,500 mg

Starch 980 mg

After mixing the above components, they are prepared so as to be 2 to 3 g per one ring according to a conventional method for producing a pellet.

Manufacturing example  6. Preparation of injections

Cymbidium extract 10 mg

180 mg mannitol

Sterile sterilized drinking water 2,970 mg

Na ₂ HPO ₄ .12H ₂ O 30 mg

(2 mL) per 1 ampoule according to a conventional injection preparation method.

Manufacturing example  7. Liquid  Produce

Cymbidium extract 10 mg

10,000 mg per isomerization

Mannitol 5,000 mg

Purified water quantity

Dissolving the above components in purified water according to a usual liquid preparation method, adding an appropriate fragrance, filling the bottle and sterilizing it.

Manufacturing example  8. Manufacture of soap

330 mL of water and 175 g of NaOH were mixed and completely dissolved. Then, 10 mg of cymbidium extract was added to the mixture in small portions for about 30 minutes. The mixture was allowed to dry in a shady, windy environment until dry.

Manufacturing example  9. Manufacture of bath agent

10 mg of cymbidium extract was added to about 60 ° C, and then a salt solution was added to make a saturated solution. Water was evaporated. The solution was cooled at room temperature and freeze-dried by rapid freezing. The lyophilized solid was powdered to prepare a bath agent.

Manufacturing example  10. Cleansing Lotion  Produce

Cymbidium extract
0.001 v / v%
Sodium ethylenediaminetetraacetate
0.02
Paraoxybenzoic acid methyl
0.2
glycerin
4
Sodium Hyruronic Acid
2
Propylene glycol
3
Carbomer
5
Cetearyl alcohol
0.7
Glyceryl stearate
0.5
Sheer butter
One
P-hydroxybenzoic acid profile
0.1
Macadamia nut oil
One
Sesquioleic acid
0.5
Glyceryl stearate
One
Polyoxyethylene sorbitan monooleate
2
Polydensen
5
Mineral oil
20
Dimethicone
5
Stearyl dimethicone
2
Triethanolamine
0.05
Spices
Suitable amount
Pigment
Suitable amount
Purified water
Balance

These ingredients are used to prepare the cosmetic preparations for cleansing lotions according to conventional methods in the art.

Manufacturing example  11. skin  Produce

Cymbidium extract
0.001 v / v%
Paraoxybenzoic acid methyl
0.2
Sodium ethylenediaminetetraacetate
0.02
1,3-butylene glycol
0.02
Allantoin
3
Sodium hyaluronic acid seed
5
Carbomer
0.1
Cetostearyl alcohol
0.7
P-hydroxybenzoic acid profile
0.1
Sorbitan olivate
1.5
Soy lecithin
0.2
Dimethicone
0.2
Cetyl octanoate
0.2
Sheer butter
0.2
Sodium placrylate
3
Triethanolamine
0.1
Imidazolidinyl urea
0.3
Spices
Suitable amount
Pigment
Suitable amount
Purified water
Balance

These ingredients are used to make the skin according to the usual methods in the field of cosmetics for making a skin.

Manufacturing example  12. Serum  Produce

Cymbidium extract 0.001 v / v% Jojoba 5 Black Sesame 2 Sweet almond 3 This malting wax One Vitamin E One glycerin 2 Hyaluronic acid One Marine Elastin One Purified water Balance

These ingredients are used to prepare according to the usual methods in the field of cosmetic manufacturing for the production of a serum.

Manufacturing example  13. Lotion  Produce

Cymbidium extract
0.001 v / v%
Paraoxybenzoic acid methyl
0.2
Sodium ethylenediaminetetraacetate
0.02
1,3-butylene glycol
0.02
Allantoin
3
Sodium hyaluronic acid seed
5
Carbomer
0.1
Cetostearyl alcohol
0.7
P-hydroxybenzoic acid profile
0.1
Sorbitan olivate
1.5
Soy lecithin
0.2
Dimethicone
0.2
Cetyl octanoate
0.2
Sheer butter
0.2
Sodium placrylate
3
Triethanolamine
0.1
Imidazolidinyl urea
0.3
Spices
Suitable amount
Pigment
Suitable amount
Purified water
Balance

The above ingredients are used to prepare according to the conventional methods in the field of cosmetics for the production of lotions.

Manufacturing example  14. Essence manufacturing

Cymbidium extract 0.001 v / v% Allantoin 0.05 Sodium ethylenediaminetetraacetate 0.02 Triethanolamine 0.2 Sodium Hyaluronic Acid 7 Imidazolidinyl urea 0.15 Sodium polyacrylate 0.4 Carbomer 0.2 ethanol 3 Polyoxyethylene sorbitan monostearate 0.2 Paraoxybenzoic acid methyl 0.2 Spices Suitable amount Pigment Suitable amount Purified water Balance

The above ingredients are used to prepare according to a conventional method in the field of cosmetics for making essences.

Manufacturing example  15. Cream production

Cymbidium extract
0.001 v / v%
Sodium ethylenediaminetetraacetate
0.02
Allantoin
0.1
glycerin
5
Paraoxybenzoic acid methyl
0.2
Sodium Hyaluronic Acid
6
Carbomer
0.1
Cetostearyl alcohol
1.7
Polydensen
2
Squalane
2
P-hydroxybenzoic acid profile
0.1
Butylene glycol dicaprylate
3
Cetyl octanoate
5
Microcrystalline lead
0.1
Triethylpentanediol
0.1
Sheer butter
0.2
Sorbitan olivate
0.3
Cyclomethicone
0.3
Stearyl dimethicone
0.5
Imidazolidinyl urea
0.15
Spices
Suitable amount
Pigment
Suitable amount
Purified water
Suitable amount

These ingredients are used to prepare the cream according to the usual methods in the art of making cosmetics.

Manufacturing example  16. Pack manufacture

Cymbidium extract
0.001 v / v%
Sodium ethylenediaminetetraacetate
0.02
Betaine
3
Glyceryl polymethacrylate
2
Allantoin
0.1
Sodium Hyruronic Acid
2
glycerin
3
Dipropylene glycol
5
Paraoxybenzoic acid methyl
0.2
Polyvinyl alcohol
10
Polyoxyethylene sorbitan monooleate
0.9
Sesquioleic acid
0.3
Jojoba ester
2
Cetearyl alcohol
1.5
Petrolatum
0.5
Spices
Suitable amount
Pigment
Suitable amount
Purified water
Suitable amount

The ingredients are prepared according to conventional methods in the art of making cosmetics for pack manufacture.

Manufacturing example  17. Massage cream manufacturing

Cymbidium extract 0.001 v / v% Purified water Suitable amount glycerin 4.0 vaseline 3.5 Triethanolamine 0.5 Liquid paraffin 24.5 Squalane 2.5 Wax 2.1 Tocopheryl acetate 0.1 Cava Paul 1.0 Sorbitan sesquioleate 3.1 incense a very small amount antiseptic a very small amount

These ingredients are used to make the usual methods in the field of cosmetics for the manufacture of massage creams.

Manufacturing example  18. Manufacture of makeup base

Cymbidium extract
0.001 v / v%
Purified water
Suitable amount
Coated silica
One
Silica
10
Titanium dioxide
8
Zinc oxide
3
Pigment
One
Plate powder
Balance

The ingredients are used to make according to the conventional method in the field of cosmetic production for make-up base production.

Manufacturing example  19. Powder fact  Produce

Cymbidium extract 0.001 v / v% Purified water Suitable amount Mica 15 Titanium dioxide 7 Silicone oil 3 Ester oil 3 Pigment Suitable amount Spices Suitable amount Talc Balance

The powder facts are prepared according to a conventional method in the field of cosmetics for making powder facts using the above components.

Manufacturing example  20. Two way  Cake manufacturing

Cymbidium extract
0.001 v / v%
Jung Soo Su
Suitable amount
Mica
15
Titanium dioxide
12
Silicone oil
3
Ester oil
5
Pigment
Suitable amount
Spices
Suitable amount
Talc
Balance

The above ingredients are used to prepare according to conventional methods in the field of cosmetics for the manufacture of two-way cakes.

Claims (5)

Wherein the cymbidium extract is contained as an active ingredient and the cymbidium extract is contained in an amount of 0.001 to 10% by weight based on the weight of the entire pharmaceutical composition. The atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, An allergic asthma and anaphylactic shock, or a pharmaceutical composition for preventing or treating contact dermatitis.
The method according to claim 1,
Wherein said pharmaceutical composition alleviates or inhibits an inflammatory reaction, itching, or edema caused by allergic diseases or contact dermatitis, or a pharmaceutical composition for preventing or treating allergic diseases or contact dermatitis.
An allergic disease selected from the group consisting of atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, allergic asthma and anaphylactic shock comprising cymbidium extract as an active ingredient, An external preparation for skin for the prevention or treatment of dermatitis.
An allergic disease selected from the group consisting of atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, allergic asthma and anaphylactic shock comprising cymbidium extract as an active ingredient, Health functional food for improving sex dermatitis.
An allergic disease selected from the group consisting of atopic dermatitis, allergic dermatitis, allergic rhinitis, allergic conjunctivitis, allergic asthma and anaphylactic shock comprising cymbidium extract as an active ingredient, A cosmetic composition for improving dermatitis.

Images