KR20160038102A - Composition for treating, improving or preventing allergic disease containing extract of Triticum aestivum Lamarck - Google Patents

Abstract

The present invention relates to a composition for preventing, ameliorating, or treating allergic disease, containing a leaf bud extract of Triticum aestivum Lamarck as an active ingredient. More specifically, the present invention relates to a composition for preventing, ameliorating, or treating allergic disease, which exhibits superior antiallergic effects. According to the present invention, the composition inhibits infiltration of inflammatory cells and edema induced by allergic reaction, and lowers concentration of interleukin 4 (IL-4), tumor necrosis factor-alpha (TNF-α), and immunoglobulin E (IgE) in serum, while suppressing production of IL-4 which is a Th2 cytokine in spleen cells.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing, ameliorating or treating hypersensitivity immune response diseases containing as an active ingredient a leaf extract of Aspergillus oryzae,

The present invention relates to a composition for preventing, ameliorating or treating a hypersensitivity immune response disease comprising as an active ingredient an aspartame extract.

Recently, the prevalence of hypersensitivity immune response disease is continuously increasing due to westernized lifestyle, and it is known that more than 25% of the population in industrialized countries is affected by hypersensitivity immune response disease.

Hypersensitivity immune response disease is a disease in which the body reacts sensitively to immunoglobulin E (IgE) mediated by an antigenic substance known as allergen. Allergic rhinitis, asthma, food allergies, atopic dermatitis, allergic conjunctivitis, urticaria, and anaphylaxis, among others.

When specific allergens come into contact with digestive organs through the skin, respiratory or food, an abnormal immune response leads to the differentiation of B lymphocytes producing immunoglobulin (Ig) antibodies and the production of antigen-specific IgE. IgE binds to the high affinity receptor (FcεRI) of mast cells in the connective tissues or mucous membranes and cross-binds with the antigen, which activates mast cells to produce amines (eg histamine), heparin, protein It is known that proteases, leukotrienes, prostaglandins, cytokines, chemokines, etc. are released to control the differentiation and activity of various immune cells and deepen the allergic immune response.

When mast cells are activated by the exposure of allergens and the release of the mediators, the inflammatory reaction is accompanied by the allergic immune response. Increased vascular permeability leads to swelling and promotes migration of lymphocytes and immune cells to inflammatory sites. TNF-α has been reported to increase the expression of endothelial cell adhesion molecule ICAM-1, which is involved in the migration of immune cells. TNF-α, an inflammatory cytokine, plays an important role in this response.

It has been known that allergen-activated Th2 cells play an important role in the initiation and maintenance of the mechanism of allergic immune response. The cytokines secreted by Th2 cells (IL-4, IL-5, IL-13, etc.) and chemokines increase the production of IgE in B cells and the activity and migration of mast cells, basophils and eosinophils, Secretion, subepithelial fibrosis, and the like, and is involved in the accompanying tissue damage. Therefore, it is important to control IgE mediated immune response, the inflammatory response accompanying it, and Th2 cell activity in the treatment of allergy.

 Currently, antihistamines, corticosteroids and leukotriene antagonists are mainly used as anti-allergy drugs. Antihistamines are drugs that inhibit the function of H1, a receptor for histamine. In the case of classical first-generation antihistamines, it has been reported to cause drowsiness and dizziness by sedating the central nervous system. Second and third generation antihistamines selectively acting at the periphery rather than the central nervous system have been recently used These drugs are also intended to relieve symptoms rather than the underlying treatment. Corticosteroid therapy is also used as a treatment for allergic diseases. However, long-term use of Cushing's syndrome (Cushing's syndrome, such as exogenous high cortisol, causing osteoporosis, adrenal insufficiency, such as side effects are reported to cause. In addition, there is a method to gradually reduce the allergic reaction by administering small amount of allergen to allergic patients. However, this method is very inconvenient in that it may cause anaphylactic shock depending on the patient and should be performed for several years. Therefore, in order to overcome these drawbacks, it is necessary to discover natural materials having anti-allergic activity which can control IgE production and excessive immune cell activity, which are less side effects than conventional drugs, safe and fundamentally causing allergy .

Wheat leaf, which means germinated leaf from wheat ( Triticum aestivum Lamarck ), is now consumed in the form of juice or dried powder and is known as a health food. It contains vitamins A, B, C, K, calcium, potassium, iron, magnesium, sodium and 17 amino acid nutrients as well as chlorophyll, minerals and enzymes. The efficacy of bean curd improves the side effects of chemotherapy in breast cancer patients, and is known to have therapeutic effects, anti-obesity and antidiabetic effects of peripheral ulcerative colitis. In addition, eosinophil counts in the blood of patients with Mediterranean anemia, in which eosinophilia is frequently observed, are reported to be reduced when the juice of beef livers is administered. In general, eosinophilia also increases in patients with asthma allergy.

As described above, the alfalfa leaf has various pharmacological effects, but studies on the antiallergic activity of the alfalfa leaf extract have not been disclosed yet.

Korean Patent Publication No. 10-2014-0099114

In order to solve the problems of the prior art as described above, the present invention provides a method for suppressing edema and infiltration of inflammatory cells caused by an allergic reaction, comprising administering to a patient in need of such treatment an effective amount of IgE, tumor necrosis factor (TNF- 4 or IL-4) in the spleen cells and inhibit the production of IL-4, which is a Th2 cytokine, in spleen cells. The present invention also provides a composition for preventing, ameliorating or treating allergic diseases.

The present invention also provides a method for inhibiting the production of IL-4, which is a cytokine secreted by Th2 cells in spleen cells, and suppressing the synthesis of immunoglobulin E (IgE) in the body against allergic diseases mediating immunoglobulin E (IgE) And a composition for preventing, ameliorating or treating an allergic disease.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating an allergic disease comprising Triticum aestivum Lamarck leaf extract as an active ingredient.

The bean leaf extract may be extracted with at least one solvent selected from water, an alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof.

The alcohol having 1 to 4 carbon atoms is preferably at least one selected from the group consisting of methanol, ethanol, propanol, isopropanol and butanol, more preferably 30 to 90% ethanol.

It is preferable that the bean leaf extract is contained in an amount of 0.01 to 95% by weight based on the total weight of the composition.

The bean leaf extract may inhibit the production of an immunoglobulin E (IgE) antibody.

In addition, the bean leaf extract can inhibit the production of tumor necrosis factor (TNF-α) and interluxin 4 (IL-4) which cause allergic inflammation.

The allergic diseases include allergic rhinitis, asthma, allergic rhinitis, psoriasis, eczema, allergic dermatitis, atopic dermatitis, contact dermatitis, allergic conjunctivitis, allergic encephalitis, urticaria, pruritus, insect allergy, , Allergic hypersensitivity, and the like.

The present invention also provides a food composition for preventing or ameliorating an allergic disease comprising as an active ingredient a tocopherol extract.

The present invention also provides a cosmetic composition for preventing or ameliorating an allergic disease comprising as an active ingredient an extract of Aspergillus oryzae.

According to the present invention, suppression of edema and infiltration of inflammatory cells induced by allergic reactions and reduction of serum IgE, tumor necrosis factor (TNF-α) and interluxin 4 (IL-4) And inhibit the production of IL-4, a Th2 cytokine, in splenocytes. The present invention also relates to a method for inhibiting the production of IL-4, a cytokine secreted by Th2 cells in spleen cells, and is useful for the treatment of allergic rhinitis, asthma, food allergy, atopic dermatitis, (IgE) for allergic diseases mediating immunoglobulin E (IgE) such as allergic conjunctivitis, urticaria, and anaphylaxis.

1 is a graph showing an effect of suppressing ear edema in a mouse induced by OVA sensitization of a leafy leaf extract according to an embodiment of the present invention.
FIG. 2 is a photograph showing the effect of inhibiting inflammatory cell infiltration at an antigen-exposed site in OVA-sensitized mice of bean leaf extract according to an embodiment of the present invention.
FIG. 3 is a graph showing the effect of decreasing IgE concentration in mouse serum induced by OVA sensitization of Ganoderma lucidum extract according to an embodiment of the present invention. In Fig. 3, A represents total IgE and B represents the results of OVA-specific IgE.
FIG. 4 is a graph showing inhibitory effects on TNF-.alpha. And IL-4 in mouse serum induced by OVA sensitization of bean leaf extract according to an embodiment of the present invention. In Fig. 4, A represents TNF-alpha and B represents IL-4.
FIG. 5 is a graph showing the effect of bean leaf extract on cytotoxicity of mouse spleen cells induced by OVA sensitization according to an embodiment of the present invention.
FIG. 6 is a graph showing the effect of Ganoderma lucidum extract on Th2 cytokines of mouse spleen cells induced by OVA sensitization according to an embodiment of the present invention. In Fig. 6, A shows the results of IL-4, B shows IL-5 and C shows the results of IL-13.
FIG. 7 is a graph showing the effect of the leaf-and-leaf extract on symptoms of dermatitis such as a mouse causing atopic symptoms according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

In the present specification, the term "wheat leaf " is meant to include all the young leaves or shoots (wheat or wheat) grown from wheat seeds to mature leaves.

The inventors of the present invention conducted a study on the development of new drugs for the prevention and treatment of diseases of allergy. As a result, the inventors of the present invention have found that the extract of Asanovorax alfalfa, The present inventors completed the present invention based on the findings that they inhibited invasion, decreased the concentrations of IgE, TNF-α, and IL-4 in serum, suppressed the production of IL-4 in spleen cells and had excellent antiallergic effects .

The present invention is characterized by containing a leafy leaf extract as an active ingredient.

The tofu leaf extract is extracted from small buds germinated from Triticum aestivum Lamarck . The tofu leaf extract can be extracted, separated and fractionated from natural extracts by using extraction, separation and fractionation methods known in the art Can be used. The 'extract' as defined in the present invention is obtained by extraction treatment from bean leaves using an appropriate solvent and includes, for example, crude extracts of Wheat leaves, polar solvent-soluble extracts and non-polar solvent-soluble extracts.

The bean leaf extract may be extracted according to various extraction solvents and extraction methods. Any suitable pharmaceutically acceptable organic solvent may be used as an appropriate solvent for extracting the extract from bean leaves, and water or an organic solvent But is not limited thereto. Examples of the solvent include water, alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol, isopropanol, and butanol, etc., Can be mixed and used. Particularly, it is preferable to use 30 to 90% ethanol (alcohol) as the solvent.

After the extract is obtained using water or an organic solvent as described above, a liquid material can be obtained by freezing, heating and filtering at room temperature by a conventional method known in the art, or a liquid material can be obtained by further evaporating, spray drying or freeze- You may.

In addition, the Gamma leaf extract, which is included as an active ingredient in the composition of the present invention, may be prepared in powder form by an additional process such as vacuum distillation, freeze drying, spray drying or the like. The extract can also be obtained as an additional purified fraction using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like .

Therefore, the bean leaf extract used in the present invention is a concept including all the extracts, fractions and purified products obtained in each step of extraction, fractionation or purification, their diluted solutions, concentrates or dried products.

Whey leaf extract, which is the active ingredient of the present invention extracted or fractionated as described above, has an effect of suppressing edema and infiltration of inflammatory cells caused by allergic reactions in an ovalbumin (OVA) sensitized allergy mouse model, (TNF-a) and interleukin-4 (IL-4), which are tumor necrosis factors.

Therefore, the bean leaf extract of the present invention can be used as a pharmaceutical, cosmetic or food composition useful for prevention, improvement or treatment of allergic diseases.

It is a disease in which the human body sensitively reacts with immunoglobulin E (IgE) mediated to an antigenic substance known as a general allergen of the above-mentioned allergic disease. Specific examples thereof include allergic rhinitis, asthma, allergic rhinitis, psoriasis, eczema, Allergic conjunctivitis, allergic encephalitis, urticaria, pruritus, insect allergies, food allergies, drug allergies, allergic hypersensitivity (anaphylaxis), and the like.

The present invention provides a pharmaceutical composition for the prevention or treatment of an allergic disease comprising as an active ingredient an aspartame extract.

The bean leaf extract is preferably contained in an amount of 0.01 to 95% by weight, more preferably 1 to 80% by weight based on the total weight of the composition. If the content is less than 0.01% by weight, the efficiency of taking may be lowered. If the content is more than 95% by weight, formulation may be difficult.

When used in the preparation of pharmaceutical compositions, the compositions of the present invention may further comprise suitable carriers, excipients and diluents commonly used. In addition, it can be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations such as ointments and creams, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. 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. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The present invention relates to pharmaceutical compositions comprising an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, RTI ID = 0.0 > effective < / RTI > amount. It will be apparent to those skilled in the art that the therapeutically effective dose and frequency of administration for the pharmaceutical compositions of the present invention will vary with the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. For a desired effect, the pharmaceutical composition of the present invention may be administered at a dose of 1 to 10,000 mg / kg / day, or may be administered once a day or divided into several doses.

The pharmaceutical compositions of the present invention can be administered to a subject in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

In addition, the composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy or biological response modifiers for the prevention or treatment of allergic diseases.

The present invention also provides a food composition for preventing or ameliorating an allergic disease, which comprises an extract of Aspergillus oryzae as an active ingredient. When the beeswax extract of the present invention is used as a food additive, it may be suitably used according to a conventional method such as adding the beeswax extract directly or mixing with another food or food ingredient.

In addition, the amount of mixture of the active ingredient, Wheat Leaf Extract, can be suitably changed according to the purpose of use (preventive, health or therapeutic treatment), and in the production of food or beverage, , Preferably 0.01 to 95% by weight, more preferably 0.01 to 15% by weight, based on the total weight of the composition. However, when it is intended for health and hygiene purposes or for the purpose of controlling health, it can be added in an amount below the above range, and there is no problem in terms of safety. Therefore, the active ingredient can be used in an amount exceeding the above range have.

There is no particular limitation on the kind of the food. Examples of the food to which the bean leaf extract of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, Tea, a drink, an alcoholic beverage, a vitamin complex, and the like.

When the food composition of the present invention is prepared as a beverage, it may contain additional ingredients such as various flavors or natural carbohydrates such as ordinary beverages. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame, and the like. The natural carbohydrate is contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , Carbonating agents used in carbonated beverages, and the like. In addition, the compositions of the present invention may include flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of the above additives is not limited to a great extent, but may be in the range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.

The present invention also provides a cosmetic composition for preventing or ameliorating allergic diseases, comprising as an active ingredient, an extract of Aspergillus oryzae.

It is preferable that the Gamma leaf extract is contained in an amount of 0.0001 to 30% by weight, more preferably 0.001 to 15% by weight based on the total weight of the composition. If the content is less than 0.0001% by weight, the effect can not be attained. If the content is more than 30% by weight, the feeling may be deteriorated.

As described above, the cosmetic composition of the present invention comprising the aspartame extract as an active ingredient can be used as a conventional auxiliary agent such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, and the like, which are commonly used in cosmetic compositions, May be further included. For example, the cosmetic composition may further contain an auxiliary component such as glycerin, butylene glycol, polyoxyethylene hardened castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, allantoin and the like .

Since the cosmetic composition of the present invention is basically applied to the skin, it can be prepared into any formulation conventionally produced by referring to the cosmetic composition of the related art. For example, they may be formulated as solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays, But is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritive cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a mask pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, the carrier component may include an animal oil, vegetable oil, wax, paraffin, starch, tracer, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, have.

When the formulation of the present invention is a powder or a spray, the carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and the like. Particularly in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane, dimethyl ether, and the like.

When the formulation of the present invention is a solution or an emulsion, the carrier component may include a solvent, a solubilizing agent, and an emulsifying agent. Specific examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.

When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol, propylene glycol or the like as a carrier component; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, trachant, and the like.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and the like.

Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.

Example 1. Preparation of bean leaf extract

Triticum aestivum Lamarck was supplied by the National Institute of Food Science and Technology , and the wheat was germinated at a constant temperature (20 ± 2 ° C) on sterile organic germinating pitermus. The sprouts were harvested at the time of cultivation for 2 ~ 3 weeks after germination, lyophilized, pulverized to a certain size and pulverized. 30 g of the powdered bead leaf frozen powder sample was ultrasonically extracted by heating with 70% ethanol for 1 hour, and then filtered and concentrated under reduced pressure using a rotary vacuum concentrator (N-1000, EYELA, Tokyo, Japan). Then, lyophilized lyophilized extract was obtained by lyophilization and completely dried. For subsequent experiments, the lyophilized extract was stored at 4 ° C until the experiment.

Experimental Example 1. Effect of suppressing allergic edema and inflammatory cell infiltration

1-1. OVA sensitized mouse model construction

The animals used in the experiment were a 6-week-old BALB / C female mouse from Samtako (Osan, Korea). The animals were housed in an experimental animal room environment with a temperature of 22 ± 2 ℃, a humidity of 55 ± 10% Adapted. After 2 weeks, when mice were 8 weeks old, a total of 25 mice of 18 to 20 g were divided into 5 mice per group, and then the mice were divided into normal group, normal control group (OVA-sensitized), experimental control group (Dexamethasone) (100mg / kg / day) and high concentration group of leaf extract (200mg / kg / day). 20 μg of ovalbumin (OVA) (grade V; Sigma-Aldrich, St Louis, MO) and 1 mg of aluminum hydroxide (Imject alum, Thermo Scientific, Cramlington, UK) were dissolved in 100 μl of phosphate buffered saline . After 2 weeks, the same proportion of yolk albumin and aluminum hydroxide solution was intraperitoneally injected into the remaining four groups except for the normal group. In the normal group, only the PBS solution was injected first and second. The bean leaf extract was mixed with 1% CMC (caeboxymethyl cellulose) solution for 13 days from the second sensitized day to 100 mg / kg and 200 mg / kg, and the Dexamethasone was 0.5 mg / Orally once a day at a fixed time. This experiment was conducted in accordance with ethical rules with the approval of Chonbuk National University Animal Experimental Ethics Committee.

1-2. Ear edema response measurement

In order to confirm the effect of suppressing allergic edema, 20 μl of 0.1% OVA solution was injected intradermally into the ear tissues of OVA-sensitized mice as described above, and the change of mouse ear thickness with time was measured with a Thickness gage , And the results are shown in Fig.

As shown in FIG. 1, the ear thickness was significantly increased after 6 hours and 24 hours in the positive control group induced by allergen induced by OVA. However, in the low-concentration and high concentration group, The results showed that the thickness of ear thickness decreased with time depending on the concentration, and it was confirmed from the result that the active ingredient of the present invention, Wheat Leaf Extract, reduced the allergic edema response.

1-3. Inhibition of inflammatory cell infiltration

As in Experimental Example 1-2, the ear tissues of mice injected with OVA were cut after 24 hours and fixed with 4% formaldehyde solution. The fixed tissue was embedded in paraffin to make a block, and then the tissue was sectioned to a thickness of 4 탆 using a pedestrian machine. Hematoxylin & Eosin staining was performed on the tissue sections, and the inflammatory cells and tissue changes were observed with an optical microscope. The results are shown in FIG.

As shown in Fig. 2, the immunoreactivity of the immunoreactive cells was significantly increased in the positive control group, which induced allergy compared with the normal group. However, in the mice administered with the low-concentration and high-concentration beef leaf extract, Dependent immune cell infiltration.

Experimental Example 2. IgE Inhibitory Effect in Serum

In Experimental Example 1-1, OVA-sensitized mice were sacrificed by cervical dislocation under ether anesthesia, and blood was collected from the abdominal aorta. The collected blood was allowed to stand at room temperature for 30 minutes, centrifuged at 3,000 rpm for 10 minutes to obtain serum, and total IgE and OVA-specific IgE were analyzed by ELISA, and the results are shown in FIG.

As shown in FIG. 3, the total IgE and OVA-specific IgE concentrations of the allergen-induced positive control group were higher than those of the normal group. In the group administered with the low-concentration and high-concentration beef leaf extract- It was confirmed that the concentration of IgE was significantly decreased.

Experimental Example 3: Inhibitory effect of TNF-a and IL-4 cytokines in serum

TNF-α and IL-4 were measured by ELISA kit (BD Biosciences) in the serum isolated as in Experimental Example 2, and the results are shown in FIG.

As shown in FIG. 4, the extract of Asanovirus L., which is an active ingredient of the present invention, produced TNF-α and IL-4 in the serum of an allergen-induced experimental animal compared with the normal control group and the allergen- , Respectively.

Experimental Example 4. Cell Viability Analysis

4-1. Culture of spleen cells

In Experimental Example 1-1, spleens were extracted from OVA-sensitized mice, and the cells were separated into single cells by filtering with a cell strainer, and erythrocytes were then removed. The separated cells were washed with phosphate buffered saline and then suspended in RPMI1640 (Thermo Scientific Hyclone (R)) containing 10% fetal bovine serum and 1% penicillin (100 units / ml) -streptomycin ) Medium at 37 ° C in a 5% CO ₂ incubator. The cells were stabilized and used in each experiment.

4-2. Cytotoxicity Assessment

In order to evaluate the cytotoxicity of Aspartic Acid Extract, the active ingredient of the present invention, LDH assay was performed and CytoTox96 ^® Non-Radioactive Cytotoxicity Assay kit was used. The splenocytes were streaked in 48-well plates at a concentration of 5 × 10 ⁵ cells / ml, and the spleen cells cultured in the above-mentioned 4-1 were streaked and stabilized. Then, the leaves of Asanovorax were removed at 0, 10, 25, 50, 100, Ml, and cultured for 24 hours. Cells were lysed by adding 20 μl of lysis solution to each positive well 45 minutes before collection of the cell culture, followed by 45 minutes of incubation at 37 ° C in a 5% CO ₂ incubator. Then, the cell culture solution was placed in an EP tube, centrifuged at 6,000 rpm for 5 minutes, and only the cell culture medium except for the cells was taken. 50 μl of the substrate solution was dispensed into each well, and light was blocked at room temperature for 30 minutes. Thereafter, 50 쨉 l of a stationary solution was added to each well to stop the reaction, the absorbance was measured at 490 nm, and the cytotoxicity was evaluated according to the following formula (1). The results are shown in Fig.

[Equation 1]

Cytotoxicity (%) = (absorbance of sample-treated group / total absorbance) x 100

As shown in FIG. 5, it was confirmed that the tocopherol extract of the present invention has cytotoxicity of less than 10% at a concentration of 100 μg / ml and does not affect cell survival rate.

Experimental Example 5. Th2 cytokine inhibitory effect of splenocytes

The spleen cells cultured in 4-1 were divided into 48-well plates at 5 × 10 ⁵ cells / ml and stabilized for 2 hours. Then, the leaf-leaf extract was treated at 0, 1, 10, and 100 μg / After stimulation with 100 μg / ml of OVA, the cells were cultured for 48 hours. The cultures were collected and the concentrations of IL-4, IL-5 and IL-13, which are Th2 cytokines, were measured using an ELISA kit (eBioscience, San Diego, CA, USA) Respectively.

As shown in FIG. 6, when the splenocytes isolated from the mice induced by OVA allergy were treated with OVA, the Th2 cytokines IL-4, IL-5 and IL-13 were significantly increased, Was significantly reduced by the leaves of Aspergillus oryzae.

Experimental Example 6. Inhibitory effect on atopic symptoms

At 8 weeks of age, the skin of the back of the BALB / C female mouse was epilated and then 1% DNCB (2,4-Dinitrochlorobenzene) solution was applied for 4 days to induce atopic dermatitis. The application was stopped for 3 days, and then 0.5% DNCB solution was applied to the skin of the mice for 7 times at intervals of 2 days for 12 days so that atopic dermatitis persisted. The bean leaf extract of Example 1 was mixed with vaseline at a low dose (1 mg) and a high dose (2 mg) in the experimental group, and then applied daily from the application of 0.5% DNCB solution to the end of the experiment. In the normal group (normal), nothing was treated after the hair removal, and only the vaseline was applied to the atopic group (DNCB). The symptoms of atopy appearing on the lesion of each group were visually observed, and the results are shown in Fig.

As shown in Fig. 7, the skin of the normal group was very clean, but various atopic symptoms such as erythema, edema, scurvy, pharyngeal fasciculation, and folliculitis were clearly observed in the atopic group. However, it was confirmed visually that dandruff extract of the present invention, which is an effective ingredient of the present invention, was significantly inhibited from dermatitis symptoms in the experimental group in which the low dose (1 mg) and the high dose (2 mg) were applied.

Formulation Example 1. Preparation of pharmaceutical preparations

Acid production

20 mg of leaf extract, 100 mg of lactose, and 10 mg of talt were mixed and filled in airtight bags to prepare powders.

Tablet manufacture

10 mg of L. leaf extract, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate were mixed and tableted according to a conventional preparation method.

Capsule preparation

According to a conventional capsule preparation method, 10 mg of the leaf extract, 3 mg of crystalline cellulose, 14.8 mg of lactose and 0.2 mg of magnesium stearate were mixed and filled in gelatin capsules to prepare capsules.

Injection manufacturing

(2 mL) of bean leaf extract, 180 mg of mannitol, 2,974 mg of sterile distilled water for injection, and 26 mg of Na ₂ HPO ₄ .2H ₂ O according to the usual injection preparation method.

Liquid preparation

According to a conventional method for preparing a liquid preparation, 20 mg of leaf extract, 10 g of isomerized sugar and 5 g of mannitol were added to purified water and dissolved, and the lemon flavor was added in an appropriate amount. Then, purified water was further added thereto, adjusted to a total volume of 100 mL, filled in a brown bottle, and sterilized to prepare a liquid preparation.

Formulation Example 2. Preparation of food preparation

Health food manufacturing

Vitamin A acetate, 70 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 g of vitamin B12, 10 mg of vitamin C, 10 g of biotin, 10 g of nicotinic acid amide 1.7 mg, folate 50 g, calcium pantothenate 0.5 mg, an appropriate amount of inorganic mixture, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium phosphate monohydrate 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 mg , Calcium carbonate (100 mg) and magnesium chloride (24.8 mg), granules were prepared, and a health food was prepared according to a conventional method. At this time, although the composition ratio of the vitamin and mineral mixture is relatively mixed with the ingredient suitable for health food, it may be arbitrarily modified.

Health drink manufacturing

According to a conventional method for producing healthy beverages, 100 mg of Ganoderma lucidum extract, 15 g of vitamin C, 100 g of vitamin E (powder), 19.75 g of iron lactate, 3.5 g of zinc oxide, 3.5 g of nicotinic acid amide, 0.2 g of vitamin A, 0.3 g of vitamin B2 and a predetermined amount of water were mixed and heated at 85 DEG C for about 1 hour with stirring. The resulting solution was filtered and sterilized in a 2 L container to be sterilized by sealing, and then stored in a refrigerator to prepare a health drink. At this time, although the composition ratio of the ingredients suitable for the beverage is comparatively mixed, the mixture ratio may be arbitrarily varied according to the demand, the demanded country, the intended use, and the regional or national preference.

Formulation Example 3. Preparation of cosmetic preparation

Flexible longevity manufacturing

0.1% by weight of the leaf extract, 5.2% by weight of 1,3-butylene glycol, 1.5% by weight of oleyl alcohol, 3.2% by weight of ethanol, 3.2% by weight of polysorbate 20, 2.0% by weight of benzophenone- 1.0% by weight of glycerin, 3.5% by weight of glycerin, a small amount of fragrance, a small amount of preservative, and a remaining amount of purified water were mixed to prepare a softening water by a conventional method.

Milk lotion manufacturing

A mixture of 0.1% by weight of Ganoderma lucidum extract, 5.1% by weight of glycerin, 4.2% by weight of propylene glycol, 3.0% by weight of tocopheryl acetate, 4.6% by weight of liquid paraffin, 1.0% by weight of triethanolamine, 3.1% by weight squalane, 2.5% by weight of macadamia nut oil, A mixture of 1.6% by weight of Solvite 60, 1.6% by weight of sorbitan sesquioleate, 0.6% by weight of propylparaben, 1.5% by weight of carboxyl vinyl polymer, a small amount of fragrance, a small amount of preservative and a remaining amount of purified water were mixed, .

Nutrition cream manufacturing

A mixture containing 0.5% by weight of leaf extract, 4.0% by weight of glycerin, 3.5% by weight of petroleum jelly, 2.1% by weight of triethanolamine, 5.3% by weight of liquid paraffin, 3.0% by weight squalane, 2.6% by weight of beeswax, 5.4% by weight of tocopheryl acetate, 3.2% by weight, carboxyl vinyl polymer 1.0% by weight, sorbitan sesquioleate 3.1% by weight, trace amount of fragrance, trace amount of preservative and remaining amount of purified water were mixed to prepare nutritional cream by a conventional method.

Massage cream manufacturing

A mixture of 0.5% by weight of leaf extract, 4.0% by weight of glycerin, 3.5% by weight of petroleum jelly, 0.5% by weight of triethanolamine, 24.0% by weight of liquid paraffin, 3.0% by weight of squalane, 2.1% by weight of beeswax, 0.1% by weight of tocopheryl acetate, A massage cream was prepared by mixing 2.4% by weight of a carboxyvinyl polymer, 1.0% by weight of a carboxylvinyl polymer, 2.3% by weight of sorbitan sesquioleate, a small amount of fragrance, a small amount of preservative and remaining water.

Body Cleanser for Cleansing

A cleansing body cleanser was prepared by mixing 1 g of leaf extract, 18 g of an anionic surfactant, 5 g of a nonionic surfactant, 7 g of glycerin, 3 g of sodium chloride, 1.5 g of natural olive liquid soap, 1 g of a perfume and 100 g of water.

Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

Claims (10)

A pharmaceutical composition for preventing or treating an allergic disease comprising Triticum aestivum Lamarck leaf extract as an active ingredient. The method according to claim 1,
The pharmaceutical composition for preventing or treating an allergic disease according to claim 1, wherein the leaf extract is extracted with at least one solvent selected from the group consisting of water, an alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof. 3. The method of claim 2,
Wherein the alcohol having 1 to 4 carbon atoms is at least one selected from the group consisting of methanol, ethanol, propanol, isopropanol, and butanol. 3. The method of claim 2,
Wherein the alcohol having 1 to 4 carbon atoms is 30 to 90% ethanol. The method according to claim 1,
The pharmaceutical composition for the prevention or treatment of allergic diseases according to claim 1, wherein the vinegar leaf extract is contained in an amount of 0.01 to 95% by weight based on the total weight of the composition. The method according to claim 1,
The pharmaceutical composition for preventing or treating an allergic disease according to any one of claims 1 to 3, wherein the tocopherol extract inhibits the production of an immunoglobulin E (IgE) antibody. The method according to claim 1,
The present invention relates to a method for preventing or treating allergic diseases, which is characterized by inhibiting the production of tumor necrosis factor (TNF-α) and interluxin 4 (IL-4) which induce allergic inflammation A pharmaceutical composition. The method according to claim 1,
The allergic diseases include allergic rhinitis, asthma, allergic rhinitis, psoriasis, eczema, allergic dermatitis, atopic dermatitis, contact dermatitis, allergic conjunctivitis, allergic encephalitis, urticaria, pruritus, insect allergy, And allergic hypersensitivity. ≪ RTI ID = 0.0 > 21. < / RTI > A food composition for preventing or ameliorating an allergic disease comprising as an active ingredient an extract of Aspergillus oryzae. A cosmetic composition for preventing or ameliorating an allergic disease comprising an extract of Aspergillus oryzae as an active ingredient.

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