KR20200082586A - A composition for preventing or treating an allergic disease, comprising a mixture of fish oil and coral extract - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating allergic diseases, containing a mixture of fish oil and a coral extract, wherein the composition inhibits the secretion of granules of mast cells, and inhibits the secretion of chemokines and cytokines, thereby being able to be used as a pharmaceutical composition, a food composition, a cosmetic composition, a quasi-drug composition, and a skin external composition for alleviating, preventing or treating allergic diseases.

Description

A composition for preventing or treating an allergic disease, comprising a mixture of fish oil and coral extract}

The present invention relates to a composition for the prevention or treatment of allergic diseases, comprising a mixture of fish oil and coral extract.

Recently, allergic diseases are on the rise, and research and development of treatments for these are also underway. The allergic disease is a disease caused by an immune hypersensitivity reaction, and includes atopic dermatitis, allergic rhinitis, and allergic conjunctivitis.

The atopic dermatitis is a complex skin disease accompanied by changes in skin dryness, itchiness, infection, skin tissue abnormalities as the disease progresses due to an abnormal allergic immune response, and is known as a chronic skin disease with chronic and recurrent course have. Atopic dermatitis progresses genetically, environmentally, and immunologically, and is particularly caused by various cells (keratinocytes, T lymphocytes, white blood cells, etc.) and factors (cytokines, immunoglobulins).

The atopic dermatitis is caused by many factors such as activation of T lymphocytes, abnormalities in the cytokine system, immunological abnormalities such as increased IgE and physiological factors, and biochemical defects in the skin. In order to treat such atopic dermatitis, it is typically treated with drugs such as steroids, including moisturizing the dried skin.

The allergic conjunctivitis is an allergic disease that refers to an inflammatory disease of the conjunctiva caused by an allergic reaction caused by a specific allergen, and allergens such as pollen in spring, dust in the air, and dandruff from animals come into contact with the conjunctiva of the eye. When allergic reactions through immune cells such as mast cells, eosinophils or basophils are triggered, various inflammatory substances such as histamine, inflammatory cytokines and inflammatory lipid metabolites (Prostaglandins & Leukotrienes) are secreted and inflammation of the conjunctiva It causes a reaction.

The allergic rhinitis is the most common allergic disease, and its incidence is gradually increasing, which affects the quality of life, and is a social and economic problem due to a decrease in production capacity and an increase in the cost of treatment. It is not yet there.

Against this background, the present inventors have tried to develop a drug that can effectively treat allergic diseases, and is derived from natural aquatic nature to secure safety.

Republic of Korea Registered Patent Publication No. 10-0990195

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of allergic diseases, comprising a mixture of fish oil and coral extract.

In addition, another object of the present invention is to provide a food composition for preventing or improving allergic diseases, comprising a mixture of fish oil and coral extract.

In addition, another object of the present invention is to provide a cosmetic composition for preventing or improving allergic diseases, comprising a mixture of fish oil and coral extract.

In addition, another object of the present invention is to provide a quasi-drug composition for preventing or improving allergic diseases, comprising a mixture of fish oil and coral extract.

In addition, another object of the present invention is to provide a composition for external application for skin for preventing or treating allergic diseases, comprising a mixture of fish oil and coral extract.

In order to achieve the object of the present invention as described above, in one aspect the present invention provides a pharmaceutical composition for the prevention or treatment of allergic diseases, comprising a mixture of fish oil and coral extract.

The term "fish oil" of the present invention is manufactured by fermenting fish by-products, and is fermented by adding microorganisms and energy sources to by-products generated in the fish processing process.

In the present invention, the "fish by-product" means a by-product generated in the process of processing or cooking fish, for example, head, tail, fin, intestine, shell, and bone. Fish derived from the "fish by-products" are not limited to this, as long as they do not contain harmful components, but the fish may be one or more selected from the group consisting of mackerel, oxfish, blowfish, halibut and squid.

The microorganism may use yeast or lactic acid bacteria, and in the case of yeast, may be yeast from the genus Saccharomyces, specifically, Saccharomyces rouxii, Saccharomyces cereviciae, One or more selected from the group consisting of Saccharomyces oviformis and Saccharomyces steineri can be used, and more specifically, Saccharomyces cerevisiae cereviciae) can be used.

For the lactic acid bacteria, Lactobacillus plantarum, Lactobacillus delbrueckii, Lactobacillus bulgaricus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus casei, Lactobacillus brevis Lactobacillus acidophilus, Pediococcus pentosaceus, Pediococcus cerevisiae, Lactococcus lactis, Leconococcitrium ), Leconostoc mesenteroide (Leuconostoc mesenteroides) and Bifidobacterium bifidum (Bifidobacterium bifidum) may be used one or more selected from the group consisting of, and more specifically, Lactobacillus plantarum (Lactobacillus plantarum) Can be used.

The term "coral" in the present invention is "Resin cockroach ( Dendronephthya sp.)". The genus of resinous cockscomb is distributed on the coast of Jeju Island and is mainly found in rock subtidal zones around 5m in depth, and may grow to a maximum height of 30cm or more. The colony is a mass of a single mass, and has tentacles bundles reminiscent of the thorns of a cactus. The color of the colony has many variations, so it has various colors such as yellow, red, pink, etc., and mainly prefers where the currents are somewhat strong.

In the present invention, the pharmaceutical composition may be applied to extraction of dried or crushed resinous cockscomb ( Dendronephthya sp.). In the present invention, the term "extract" is an extract obtained by an extraction treatment, a diluent or a concentrate of the extract, a dried product obtained by drying the extract, a crude product or a purified product of the extract, or mixtures thereof, the extract itself and the extract Contains extracts of all formulations that can be formed using.

In the present invention, drying of the extract may be performed by a known method in a range in which useful components are not destroyed, and may be performed by, for example, a method of natural drying in shade. In addition, crushing or pulverization may be pulverized or pulverized to a powder sufficient to extract useful components in the subsequent extraction process. The drying and crushing or grinding processes may be performed by reversing the order as necessary or repeatedly.

In the extraction of the present invention, the extraction method is not particularly limited, and can be extracted according to a method commonly used in the art.

The term "resin cockscomb ( Dendronephthya sp.)" of the present invention means an extract obtained by extracting the resinous cockscomb ( Dendronephthya sp.). The resin in cockscomb (Dendronephthya sp.) Extract can be obtained in resin cockscomb (Dendronephthya sp.) By extraction using water, an organic solvent, or a mixed solvent thereof. Specifically, Dendronephthya sp. extract is used in the resin cockscomb as an elution solvent using alcohols having a carbon number of 1 (C1) to 4 (C4) such as water, methanol, ethanol, and butanol, or a mixed solvent thereof, as an elution solvent. Can be extracted using extraction methods such as hot water extraction, cold saliva extraction, reflux cooling extraction, or ultrasonic extraction at 10 to 100°C, but can be used without limitation if it is a method of extracting substances that have the prevention or treatment activity of allergic diseases. Can be.

More specifically, (1) removing foreign substances of the collected resinous male cockscomb ( Dendronephthya sp.), (2) crushing the resinous cockscomb ( Dendronephthya sp.), (3) crushed resinous cockscomb ( Dendronephthya sp.) is subjected to ultrasonic treatment, and (4) centrifugation to recover and filter the supernatant, thereby obtaining an extract of the genus Resin.

The step (1) is a pre-treatment step of the sample, and is a washing step to remove as much as possible the foreign substances in the resin cockscomb ( Dendronephthya sp.) collected in running water. Step (2) is a step of crushing the washed resin cockscomb genus ( Dendronephthya sp.), which is a step of crushing the resin cockscomb genus ( Dendronephthya sp.) into a blender and pulverizing it into a thick liquid. The step (3) is a step of ultrasonicating the crushed resin cockscomb ( Dendronephthya sp.), which can be ultrasonicated by adding a solvent for smooth ultrasonic treatment. The solvent includes water, an organic solvent commonly used for extraction such as ethanol or methanol, and oils capable of extracting fat-soluble substances. The organic solvent and oils may be mixed and a single solvent may be used. The ultrasonic treatment may be performed for 1 to 30 hours at a frequency of 100 to 2000W. In the step (4), the supernatant is collected by filtration through centrifugation of the ultrasound treatment material, and filtration can be performed using a micro filter or a membrane filter, and the final filtered extract is obtained by filtration. .

The genus Resin ( Dendronephthya sp.) is a large resin cockscomb ( Dendronephthya gigantea ), a prickly resin cockscomb ( Dendronephthya spinulosa ) , a purple resin cockscomb ( Dendronephthya puetteri ) , a night resin cockscomb ( Dendronephthya castanea ) , a Dendronephthya red- eye It may be Dendronephthya suensoni ) , Dendronephthya mollis , or white resin cockscomb ( Dendronephthya alba ).

The mixture of fish oil and coral extract is prepared by mixing fish oil and coral extract, and the fish oil and coral extract may be mixed in a weight ratio (w/w) of 1-99: 99-1, more specifically 1: It may be mixed in a weight ratio (w/w) of 1 to 1:4. A solubilizer may also be used for mixing the fish oil and coral extract, and the solubilizer may be a solubilizer commonly used in food or cosmetics.

In the present invention, the term "allergic disease" means a disease caused by an immune hypersensitivity reaction, and an allergic reaction is a complex reaction involving various cells and mediators and functions as a cause of various diseases. Specifically, the allergic disease may be at least one selected from the group consisting of atopic dermatitis, allergic rhinitis and allergic conjunctivitis, but is not limited thereto.

In one embodiment of the present invention, as a result of treating a mixture of fish oil and coral extracts on mast cells inducing allergic reactions using IgE and BSA, beta-hexosaminidase (β-hexosaminidase, degranulated living body) from mast cells It was confirmed that it suppressed the secretion of the marker) in a concentration-dependent manner, and it was confirmed that the gene expression of TARC, a chemokine, was decreased in a concentration-dependent manner by the treatment of the mixture, and the inflammatory cytokines IL-4, IL-5, and IL- 13, by significantly reducing the mRNA expression of IFN-γ, and TNF-α, confirming that it can be used as a composition for preventing or treating allergic diseases such as atopic dermatitis, allergic rhinitis, and allergic conjunctivitis (Figs. 2 to 5).

In the present invention, "prophylaxis" refers to all actions that suppress the symptoms of inflammation or delay the onset of symptoms by administration of the composition, and "treatment" means that all symptoms of inflammation caused by the administration of the composition are improved or beneficially altered. It means act.

In the present invention, the term "administration" refers to the introduction of the pharmaceutical composition of the present invention to an individual by any suitable method, and the route of administration of the composition is through oral or parenteral various routes as long as it can reach the target tissue. It may be administered, specifically, oral, rectal, topical, intravenous, intraperitoneal, intramuscular, intraarterial, transdermal, intranasal, inhalation or intradermal routes.

Pharmaceutical compositions comprising a mixture of fish oil and coral extracts of the present invention may further include suitable carriers, excipients or diluents commonly used in the manufacture of pharmaceutical compositions.

The pharmaceutical composition is any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, intravenous solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-driers and suppositories. It may have a formulation of, it can be a variety of oral or parenteral formulations. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin. It can be prepared by mixing the back. In addition, lubricants such as magnesium stearate, talc and the like may be used in addition to simple excipients. Liquid preparations for oral administration include suspending agents, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" in the present invention means an amount sufficient to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level refers to the individual type and severity, age, sex, and disease. It can be determined according to the type, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and discharge, the duration of treatment, the factors including the drugs used simultaneously, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, and can be easily determined by those skilled in the art.

The pharmaceutical composition of the present invention is not particularly limited as long as it is an individual for the purpose of treating allergic diseases, and any of them can be applied. For example, any of non-human animals such as monkeys, dogs, cats, rabbits, mormots, rats, mice, cows, sheep, pigs, goats, humans, birds, and fish can be used, and the pharmaceutical composition is non-oral, It can be administered subcutaneously, intraperitoneally, in the lung, and intranasally, and for local treatment, it may be administered by a suitable method, including intralesional administration, if necessary. The preferred dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the individual, the severity of the disease, the drug form, the route and duration of administration, but can be appropriately selected by those skilled in the art. For example, it may be administered by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine epidural or cerebrovascular injection, but is not limited thereto.

A suitable total daily dosage can be determined by the treating physician within the correct medical judgment, generally in an amount of 0.001 to 1000 mg/kg, preferably 0.05 to 200 mg/kg, more preferably 0.1 to 100 mg/ The amount of kg may be administered once or several times a day.

In another aspect, the present invention provides a food composition for preventing or improving an allergic disease, comprising a mixture of fish oil and coral extract.

In the present invention, the description of "mixture of fish oil and coral extract" and "allergic disease" is as described above.

The food composition of the present invention may include pill, powder, granule, needle, tablet, capsule, or liquid forms, and there is no particular limitation on the type of food to which the mixture of fish oil and coral extract of the present invention can be added. . Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes.

In addition to the mixture of fish oil and coral extract, other ingredients may be added to the food composition, and the type is not particularly limited. For example, it may contain various herbal extracts, food additives, food additives or natural carbohydrates, etc., as additional ingredients, such as conventional foods, but is not limited thereto.

In the present invention, the term, "food supplement additive" refers to a component that can be added to food supplementally, it is added to prepare the food of each formulation can be appropriately selected and used by those skilled in the art. Examples of food supplements include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonic acid used in carbonated beverages, etc. are included, but the types of food supplements of the present invention are not limited by the examples.

Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; And polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol, and natural flavoring agents (such as taumatin) and stevia extracts (rebaudioside A, glycyrrhide) Hygiene, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.

The food composition of the present invention may include a health functional food. The term "health functional food" as used in the present invention refers to food prepared and processed in the form of tablets, capsules, powders, granules, liquids and pills, etc. using ingredients or ingredients having useful functionality for the human body. Here, the term "functionality" means obtaining a useful effect for health use, such as adjusting nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be manufactured by a method conventionally used in the art, and may be prepared by adding raw materials and ingredients commonly added in the art. In addition, unlike general medicines, it has the advantage that there are no side effects that can occur when taking medicines for a long time using food as a raw material, and it can be excellent in portability.

The mixing amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, the active ingredient of the present invention in the manufacture of food can be added in an amount of 1 to 50% by weight, preferably 5 to 10% by weight of the raw material composition, but is not limited thereto. However, in the case of long-term intake for health and hygiene purposes or for health control purposes, the amount may be used below the above range.

In another aspect, the present invention provides a cosmetic composition for preventing or improving allergic diseases, comprising a mixture of fish oil and coral extract.

In the present invention, the description of "mixture of fish oil and coral extract" and "allergic disease" is as described above.

On the other hand, in the cosmetic composition for preventing or improving an allergic disease of the present invention, the active ingredient may be included in any amount (effective amount) according to the purpose, formulation, compounding purpose, etc. as long as it can prevent or improve the allergic disease May be, a typical effective amount may be included in the range of 0.001% to 99.99% by weight based on the total weight of the composition. Here, "effective amount" refers to the amount of active ingredients that can induce the prevention or improvement effect of allergic diseases. Such effective amount can be determined empirically within the range of ordinary skill in the art.

The cosmetic composition for preventing or improving an allergic disease of the present invention may be prepared in various forms. For example, the cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, a solution. , Suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray, etc. may be formulated, but is not limited thereto. In addition, specifically, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, soap, shampoo, It may have a formulation selected from the group consisting of cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, lipstick, makeup base, foundation, press powder and rust powder, but is not limited thereto.

The cosmetic composition for preventing or improving an allergic disease of the present invention may include an acceptable carrier in a cosmetic preparation in addition to its active ingredient. Here, the "acceptable carrier in cosmetic preparations" is a compound or composition that is already known and used that may be included in cosmetic preparations, or is a compound or composition to be developed in the future, and has no toxicity, instability, or irritability beyond the body's adaptability when contacted with skin. Say things. The carrier is from about 1% to about 99.99% by weight, preferably from about 5% to about 99% by weight of the weight of the composition in the cosmetic composition for preventing or improving allergic diseases of the present invention. Can be included. However, since the above ratio depends on the formulation of the cosmetic preparation of the present invention and its specific application site (face or hand) or its preferred application amount, the above ratio limits the scope of the present invention in any aspect. It should not be understood.

Meanwhile, as the carrier, alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizing agents, viscous modifiers, emulsions, stabilizers, sunscreens, colorants, fragrances, and the like can be exemplified. Alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscous modifiers, emulsions, stabilizers, sunscreens, colorants, compounds/compositions that can be used as fragrances, etc., which can be used as the carrier, are already known in the art. As it is, those skilled in the art can select and use an appropriate material/composition.

In another aspect, the present invention provides a quasi-drug composition for preventing or improving an allergic disease, comprising a mixture of fish oil and coral extract.

In the present invention, the description of "mixture of fish oil and coral extract" and "allergic disease" is as described above.

As used in the present invention, the term "quasi-drug" refers to articles that are used for the purpose of diagnosing, treating, improving, alleviating, treating, or preventing diseases of humans or animals, which have less action than pharmaceuticals, for example, According to the report, quasi-drug products exclude products used for pharmaceutical use, and include products used to treat or prevent disease in humans and animals, and products that have little or no direct action on the human body.

The quasi-drug composition of the present invention is a body cleanser, disinfectant cleaner, detergent, kitchen detergent, detergent for cleaning, toothpaste, gargle, wipes, detergent, soap, hand wash, hair cleaner, hair softener, humidifier filler, mask, ointment and filter filler It can be prepared in a formulation selected from the group consisting, but is not limited thereto.

In another aspect, the present invention provides a composition for external application for skin for preventing or treating allergic diseases, comprising a mixture of fish oil and coral extract.

In the present invention, the description of "mixture of fish oil and coral extract" and "allergic disease" is as described above.

The term "external preparation" of the present invention is a preparation provided for external use, and there are external preparations, external preparations, external preparations, ointments, warning agents, suppositories, and the like. Includes.

The external preparation for skin according to the present invention may be a parenteral dosage form formulated in a solid, semi-solid or liquid form by adding a commercially available inorganic or organic carrier, excipient and diluent. The preparation for parenteral administration may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions and emulsions, but is not limited thereto.

Carriers, excipients and diluents that may be included in the external preparation include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The composition for external application for skin by each formulation can be formulated by appropriately selecting other ingredients other than the mixture of fish oil and coral extract of the present invention without difficulty by those skilled in the art according to the formulation or purpose of use of the composition for external application for skin. When applied, a synergistic effect may occur.

The composition for external application for skin of the present invention may include an active ingredient in an amount of 0.0001 to 30% (w/v) by weight of the total composition. If included below 0.0001% (w/v), the skin whitening effect cannot be expected substantially. On the other hand, if it contains more than 30% (w/v), the overall processability of the composition, such as solubility as a solvent, is poor, and thus use in various applications such as formulation may be limited.

The present invention relates to a composition for preventing, improving or treating allergic diseases, comprising a mixture of fish oil and coral extracts, inhibiting granular secretion of mast cells, inhibiting chemokine and cytokine secretion, and preventing allergic diseases. It may be used as a pharmaceutical composition for improving, preventing or treating, a food composition, a cosmetic composition, a quasi-drug composition and a composition for external application for skin.

1 shows the survival rate of BMCMC according to a mixture treatment of coral extract and fish oil.
Figure 2 is a measure of the release amount of β-hexosaminidase of BMCMC according to a mixture treatment of coral extract and fish oil.
Figure 3 measures the inhibitory efficacy of a mixture of coral extract and fish oil on the expression of inflammatory chemokine mRNA of activated BMCMC.
Figure 4 is a measure of the cytokine mRNA inhibitory effect according to the mixture treatment of coral extract and fish oil in the activated BMCMC.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example 1. Preparation of fish oil

Fish oil, which is a material for this experiment, was aged (fermented) by adding 50 Kg of each by-product generated during processing of mackerel, oxfish, blowfish, squid, and halibut into a fermentation container, and adding commercially available 10 Kg per sugar and microorganism.

Specifically, Lactobacillus plantarum (1.0Х10 ⁸ cells/cfu) and Saccharomyces cerevisiae (3.0Х10 ⁸ cells/cfu) pre-cultured in YM and MRS liquid medium were used as the microorganism. The pre-cultured microorganisms were inoculated into 2 L medium, respectively, and cultured at 30°C for 15 days. Subsequently, the microorganisms were fermented in a fermentation vessel containing by-products of fish sugar and fish by fermentation (fermentation) for 10-60 days after adding 1.0*10 ⁶ ~ 1.0*10 ¹⁰ cell/cfu, 50~5,000ml, respectively .

Hexane was added to the culture mixture after the cultivation and aging in a 3-fold volume (1:3, v/v), and the hexane layer was separated after mixing. Then, after separating the oil sample (fish oil) with a vacuum concentrator, it was placed in a glass container until it was used in the experiment and kept refrigerated.

Comparative Example 1. Preparation of fish oil

Fish oil was prepared in the same manner as in Example 1, except that microorganisms were added to the by-products of the fish.

Example 2. Preparation of coral extract

In order to remove foreign substances from the collected large resin cockscomb ( Dendronephthya gigantea ), it was washed with running water. Then, the washed large resin cockscomb ( Dendronephthya gigantea ) was put into a blender and crushed into a thick liquid. The crushed large resin cockscomb ( Dendronephthya gigantea ) was sonicated for 3-10 hours at a frequency of 500-1500w . Next, the supernatant was recovered by centrifugation of the sonication product, and filtered using a filter to obtain a filtered extract.

Example 3. Preparation of a mixture of fish oil and coral extract

A mixture of fish oil and coral extract was prepared by mixing the fermented fish oil prepared in Example 1 and the large resin cockscomb extract prepared in Example 2 in a weight ratio of 1:3.

Experimental Example 1. Measurement of cell viability

MTT assay was performed to measure the efficacy search of a mixture of fish oil and coral extract (Example 3) for the survival of BMCMC (bone marrow-derived cultured mast cells, BMCMC). BMCMC (2×10 ⁴ cells/well) was dispensed into a 96 well plate, and a mixture of fish oil and coral extract (MIX) was treated by concentration and incubated at 37° C., 5% CO ₂ incubator for 24 hours. Thereafter, 15 μL of MTT (5 mg/ml) was added to each cell, and after incubation for 4 hours in a 37°C, 5% CO ₂ incubator, 100 μL of DMSO solution (50% DMSO, 10%) SDS, pH 7.4) was treated to dissolve the formazan precipitate formed, and absorbance was measured at a wavelength of 540 nm using an ELISA reader.

As a result, as a result of measuring the cytotoxicity of the mixture of fish oil and coral extract (Example 3) in BMCMC, it showed a similar cell survival rate to the untreated control group. From this result, it was confirmed that the mixture showed no toxicity to BMCMC (FIG. 1).

Experimental Example 2. β-hexosaminidase release inhibition efficacy measurement

Measurement of the amount of β-hexosaminidase release was conducted to determine the degree of inhibition of allergic reactions at the cellular level by measuring the amount of β-hexosaminidase, an indicator of degranulation.

First, BMCMC was dispensed into e-tubes to be 2×10 ⁵ cells per tube, and 50 μl of 0.1% BSA-tyroide's buffer and a mixture of fish oil and coral extract (MIX) (Example 3) were treated by concentration. After 2 hours, anti-DNP-specific IgE was dispensed into each tube at a concentration of 2 μg/10 ⁵ cells. To stabilize BMCMC, vortexing was performed on ice for 60 minutes every 10 minutes. Thereafter, 500 µl of 0.1% BSA-tyroide's buffer was added, and after washing, the number of cells was counted at the last step for 60 minutes at 37°C for 10 minutes.

Centrifugation was performed at 2000 rpm for 3 minutes, and 50 μl of 0.5% triton 100-tyroide's buffer was added to a tube containing cells with an antigen-antibody reaction to measure β-hexosaminidase release, and then reacted for 20 minutes in ice after vortexing. The supernatant thus obtained and the substrate buffer (4-p-nitrophenyl-N-acetyl-β-D-glucosaminide 1.3 mg/ml, 0.1 M sodium citrate, pH 4.5), which is a substrate of β-hexosaminidase, were mixed in a 96 well plate and mixed with 37 After reacting for 40 minutes at ℃, the reaction was terminated by treating 0.2 M glycine (pH 10.7) per well, and absorbance was measured at a wavelength of 405 nm using a microplate reader.

Mast cells contain granules containing various allergens, and when activated by antigen, the granules fuse with the cell membrane and secrete histamine, β-hexosaminidase, tryptase, etc., in the cells, to cause type 1 allergy It is known to be the main cause of the reaction. Therefore, in order to measure the degranulation phenomenon, the inhibitory effect of the mixture of fish oil and coral extracts on β-hexosaminidase secreted out of cells by the antigen in BMCMC was confirmed.

As a result, the amount of β-hexosaminidase secretion was found to be significantly increased in the group treated with anti-DNP-IgE and DNP-BSA compared to the untreated control group that did not process anything, and the fish oil and coral When the mixture of extracts was treated together, it was confirmed that the concentration-dependently decreased β-hexosaminidase release amount than the group treated with anti-DNP-IgE and DNP-BSA at all concentrations (p<0.05) (FIG. 2 ). .

Thus, the mixture of fish oil and coral extract (MIX) (Example 3) inhibits mast cell degranulation (beta-hexosaminidase secretion) caused by IgE and BSA, thereby inhibiting activation of mast cells to have anti-allergic efficacy. Confirmed.

Experimental Example 3. Measurement of the effect of suppressing the expression of chemokine causing allergic diseases including atopic dermatitis

Chemokine represents chemoattractants known as low-molecular proteins, and can be divided into four submailies, C, CC, CXC, and CX3C, depending on the structure and function of the protein. Causes more. Among these chemokine, TARC is a representative Th2 chemokine and is known to induce the migration and infiltration of Th2 cells to the inflammatory site. Since the increase in inflammatory chemokine is very closely related to allergic diseases such as atopic dermatitis in addition to cytokine, the inhibitory effect of the mixture of fish oil and coral extract (MIX) on the mRNA expression of inflammatory chemokine is increased when inducing allergic disease. RT-PCR was performed to confirm.

As a result, it was confirmed that the gene expression of TARC, a chemokine significantly increased by stimulation of anti-DNP-IgE and DNP-BSA, was concentration-dependently decreased by the treatment of the mixture (Example 3), and anti-DNP- There was a significant difference from the group treated with only IgE and DNP-BSA (p<0.05) (FIG. 3 ).

Experimental Example 4. Measurement of the inhibitory effect of cytokine expression on allergic diseases including atopic dermatitis

When inducing allergic diseases including atopic dermatitis, RT-PCR was performed to confirm the inhibitory efficacy of the mixture against mRNA expression of increasing inflammatory cytokine. BMCMC cells were divided into 2×10 ⁶ cells, and a mixture of fish oil and coral extract (MIX) (Example 3) was treated for each concentration, cultured for 2 hours, stimulated with DNP-IgE and BSA, and cultured for 24 hours. . Then, after removing the supernatant, trizol reagent was added to the cells, chloroform was added, and vortexing was performed for 10 seconds. Thereafter, after centrifugation at 12000 rpm for 15 minutes, the supernatant was separated and mixed with isopropanol.

Next, the supernatant was removed by centrifugation at 12000 rpm for 15 minutes, and the pellet was mixed and used for RT-PCR. The isolated total RNA was synthesized using cDNA synthesis kit, and PCR products were prepared using RT-PCR kit. Each PCR product was loaded on a 1% agarose gel and analyzed through electrophoresis for 30 minutes at 100V.

As a result, compared to the control group that did not process anything, the inflammatory cytokine IL-4, IL-5, IL-13, IFN-γ, and TNF-α in the group treated with anti-DNP-IgE and DNP-BSA mRNA expression was significantly increased. On the other hand, when the mixture of fish oil and coral extract (Example 3) was treated by concentration, the inflammatory cytokine IL-4, IL-5, IL-13, IFN-γ, and It was confirmed that the mRNA expression of TNF-α was significantly reduced (p<0.05) (FIG. 4 ).

In conclusion, the mixture inhibits the degranulation of mast cells (beta-hexosaminidase secretion) and suppresses the expression of chemokine and cytokine, which are involved in inducing allergic diseases including atopic dermatitis, and suppresses the activation of mast cells, thereby antiallergic. It was confirmed to have efficacy.

Claims (8)

A pharmaceutical composition for the prevention or treatment of allergic diseases, comprising a mixture of fish oil and coral extract. The pharmaceutical composition for preventing or treating allergic diseases according to claim 1, wherein the fish oil is produced by fermenting fish by-products. According to claim 1, The coral extract is extracted with water, C ₁ -C ₄ alcohol or a mixed solvent thereof, pharmaceutical composition for preventing or treating allergic diseases. The pharmaceutical composition for preventing or treating allergic diseases according to claim 1, wherein the allergic disease is at least one selected from the group consisting of atopic dermatitis, allergic rhinitis and allergic conjunctivitis. A food composition for the prevention or amelioration of allergic diseases, comprising a mixture of fish oil and coral extract. A cosmetic composition for preventing or improving allergic diseases, comprising a mixture of fish oil and coral extract. A quasi-drug composition for preventing or improving allergic diseases, comprising a mixture of fish oil and coral extract. A composition for external application for skin for preventing or treating allergic diseases, comprising a mixture of fish oil and coral extract.

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