KR101949557B1 - Composition for immune enhancing activity containingextract of aralia cordata - Google Patents

Abstract

The present invention relates to a composition for enhancing immunity comprising an extract of Aspergillus oryzae as an active ingredient. The extracts of Corydalis did not show cytotoxicity even at a high dose, but cytotoxicity and induction of cytokine production were measured using mouse macrophage-derived macrophages, and mice were orally administered and then infected with Listeria spp. It has been confirmed that it is very excellent in enhancing effect. Therefore, it can be applied to animals and humans as an immunity enhancing composition which has recently been spotlighted as a substitute for antibiotics, and its industrial effect will be excellent.

Description

Technical Field [0001] The present invention relates to a composition for enhancing immunity,

The present invention relates to a composition for enhancing immunity comprising an extract of Aspergillus oryzae as an active ingredient, specifically a food composition for immunostimulating or an immunostimulating feed composition.

The immune system can be divided into natural resistance, nonspecific immune system and specific immune system. Natural resistance (first line of defense) is an anatomical physiological element that blocks all invaders, including microorganisms, regardless of their type. Nonspecific immunity (second line of defense) is a phagocyte that breaks through natural resistance and removes intruders. A specific immune system is a highly developed immune system with the ability to distinguish between memory and non-immune systems. The immune system is composed of lymphocytes, and the specific immune system (the third line of defense).

The leukocytes constitute a secondary or tertiary defense line, which breaks the primary line and takes charge of the foreign body. In the case of bacterial, viral infection or inflammatory reaction, the control of macrophage and lymphocyte activity is decided on the therapeutic effect of the drug It plays a pivotal role. Macrophage is a multifunctional cell that plays an important role in the immune system by producing various cytokines and NO in oxidative stress situations.

The macrophages are microbicidal cells that antigen-present, tumoricidal, or kill microbial cells, and play a central role in cell-mediated or humoral immunity The NO produced in the activated macrophages shows a macrophage function as a non-specific host defense mechanism and a proliferation inhibitory activity of bacteria and cancer cells. In particular, iNOS expressed by stimulation such as lipopolysaccharide (LPS), cytokine, and TNF-α in macrophages is known to produce large amounts of NO over a long period of time and promote NFκB activity. Reactive oxygen species (ROS) such as superoxide anion (O ₂ ^- ), hydrogen peroxide (H ₂ O ₂ ) and nitric oxide (NO) by activated macrophages Production is an important cytotoxic and cytostatic mechanism in nonspecific immunity.

In relation to immunological activity, hematopoietic stem cells are damaged in cancer treatment such as radiotherapy or chemotherapy, resulting in reduction of hematopoietic cells and immune cells, thereby often causing obstacles to hematopoiesis and formation of immune function . In recent years, it has been reported that not only a decrease in immune activity due to chemotherapy of cancer patients but also a general person who does not have a specific disease, the immunity is weakened due to environmental reasons or lifestyle.

Due to these circumstances, many plant extracts are being used as medicines, medicines or health functional foods in order to improve health through immunity enhancement at present in the world.

The plant extract capable of enhancing such immunity makes it possible to treat, prevent or ameliorate a bacterial infection, a viral infection, or a disease caused by cancer cells. Such effect is not caused by antibacterial action but rather by improvement of immune function in the body It has an advantage that it can be free from problems such as side effects. For example, Taxol, an anticancer agent obtained from the bark of Western blotch, has an advantage that side effects are not a problem as compared with a therapeutic agent derived from a specific compound.

Therefore, by selecting a substance having an immunopotentiating effect among natural products expected to minimize the possibility of a safety problem, development of an immunopotentiating ingredient having a high immunity enhancement effect on the immune enhancement and a weak side effect Is required.

10-1141314B 10-1151484B 10-1245582B

It is an object of the present invention to overcome the problems of the prior art as described above, and it is an object of the present invention to provide a composition for enhancing immunity which comprises extracts of Aspergillus oryzae as an active ingredient.

In order to achieve the above object, the present invention udo (Aralia cordata ) extract as an active ingredient. The immunoconjugate composition may be a food composition. The food composition may be a functional food composition. The functional food composition may be in the form of a powder, a granule, a tablet, a capsule or a beverage.

The extract may be extracted with at least one solvent selected from the group consisting of water and an organic solvent, preferably water. In this respect, the extract may be preferably a hydrolyzate extract.

In addition, the present invention provides an animal feed additive for immunity enhancement comprising an extract of Aspergillus oryzae as an active ingredient. The extract may be an extract of a pine nut. The present invention also provides an animal feed enhancer comprising the animal feed additive as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for enhancing immunity, comprising the extract of Aspergillus oryzae as an active ingredient. The pharmaceutical composition for enhancing immunity may be a pharmaceutical composition.

Unless otherwise specified in the present specification, an effective component means a component that provides an immunostimulating effect in a composition, specifically, a pharmaceutical composition or a food composition.

As used herein, the term " prophylactic " means any action that inhibits or slows down the progress of symptoms due to immune function depression due to administration of the composition of the present invention, for example, infectious diseases.

Unless otherwise specified in the present specification, treatment means all improvement actions that improve or ameliorate a symptom such as a disease caused by a decrease in immune function due to administration of the composition of the present invention, for example, an infectious disease.

Unless otherwise specified herein, an individual refers to a human, a monkey, a dog, a goat, a pig, a dog, a dog, a dog, a dog, a dog, a dog or the like having a disease that can alleviate a disease caused by impaired immune function, It means all animals such as mice.

Administration, as used herein, unless otherwise indicated, means providing the subject invention with a composition of the invention in any suitable manner.

As used herein, unless otherwise specified, pharmacologically acceptable means physiologically acceptable and does not cause an allergic reaction such as gastrointestinal disorder, dizziness, or the like, when administered to an animal, preferably a human, do.

Unless otherwise specified in the present specification, immune enhancement means a function of promoting an immune response to an antigen nonspecifically in the initial activation process of immune cells or enhancing immunity by increasing the activity of cells of the immune system.

As used herein, unless otherwise stated, the disease resulting from impaired immune function may be any selected from the group consisting of infectious diseases, including cold, chronic fatigue allergy disease, and cancer. Examples of the allergic diseases include atopy, asthma, and allergic rhinitis.

The inventors of the present invention have studied about a material having immunity enhancing effect that can enhance immunity through stimulation of the secretion of immunocytokines among natural substances which have few side effects and are easy to be administered, Of plant extracts, especially Aralia cordata ) did not show cytotoxicity even at a high dose, and the experiment using mouse macrophage-derived macrophages showed that the effect of promoting the production of inflammatory cytokines related to innate immunity was excellent, and the number of lymphocytes And an immunological enhancement effect of an animal model that induced Listeria infection was excellent, and confirming that the immunity enhancement effect is very excellent, the present invention has been completed.

Hereinafter, the present invention will be described in more detail.

The invention udo (Aralia cordata extract as an active ingredient. The composition for enhancing immunity may be a food composition for enhancing immunity, and the composition for enhancing immunity may be a functional food composition for enhancing immunity.

The Aralia cordata) is bulriwoomyeo known as Aralia cordata, a plant that is easy to grow yakchogwa grows in mountainous and high viability is readily available and has been used as a medicine in traditional Korean medicine long ago. Anticholinergic effects, antioxidant effects, hypocholesterolemic effects, and hypolipidemic effects have been known to date, but no known effects on the immune system are known.

The extract can be prepared by extracting with an extraction solvent or extracting with an extraction solvent and then fractionating the extract with a fraction solvent.

The extraction solvent may be at least one selected from the group consisting of water and an organic solvent, and the organic solvent is at least one selected from the group consisting of a polar solvent and a non-polar solvent containing an alcohol having from 1 to 4 carbon atoms, Is preferably at least one selected from the group consisting of water and an alcohol having from 1 to 4 carbon atoms, more preferably water. The extraction solvent may be used in an amount of 1 to 100 times, or 5 to 20 times, based on the weight of the extract.

The organic solvent may be a polar solvent such as an alcohol having 1 to 5 carbon atoms, a diluted alcohol such as ethyl acetate or acetone, a nonpolar solvent such as ether, chloroform, benzene, hexane or dichloromethane, or a mixed solvent thereof.

The method for extracting the above extract can be produced by a conventional extraction method, specifically a solvent extraction method, in the technical field of the present invention, and examples thereof include a heat extraction, a cold extraction, a reflux cooling extraction, an ultrasonic extraction and a steam extraction . In order to carry out this extraction method, it can be carried out using a conventional extracting device, an ultrasonic pulverizing extractor or a fractionator. For example, when water is used as the extraction solvent as the extraction solvent, it is preferably hydrothermal extraction. The temperature of the water is preferably from 90 to 150 ° C or from 110 to 140 ° C and from 1 to 3 kg / cm ² or from 1.5 to 2.5 kg / cm < ² > for 1 hour to 12 hours or 2 hours to 6 hours.

The extract may be prepared by performing the extraction process and then drying the extracted extract by a drying method such as hot air drying or freeze drying.

The fraction solvent for carrying out the fraction may be water, butanol, ethyl acetate, chloroform, hexane or a mixture thereof. The fraction may be an extract prepared by the above extraction method, specifically, a fraction obtained by further fractionating the crude extract. The fraction solvent may be a solvent selected from the group consisting of ethyl acetate, ether, chloroform, benzene, hexane, methylene chloride, and mixed solvents thereof. Specifically, the fractionation step is performed by sequentially adding hexane, chloroform, ethyl acetate, butanol, and water to the crude extract in this order, then sequentially fractionating the hexane fraction, the chloroform fraction, the ethyl acetate fraction, the butanol fraction, . ≪ / RTI >

After the extraction or fractionation, the extract or fraction may be subjected to a vacuum filtration process or may be further concentrated and / or lyophilized to concentrate or remove the solvent. The obtained extract can be stored in a deep freezer until use.

Specifically, the extract may be further subjected to a filtration step, a concentration step and / or a drying step after the solvent extraction step. Alternatively, the filtration step may be performed using a filter paper, The process may be carried out through reduced pressure concentration or the like, and the drying process may be performed by a method such as hot air drying or freeze drying.

The composition for immunomodulation has application effects of improving or preventing influenza during a person's season, improving or preventing bacterial diarrhea, improving or preventing bacterial infection, and enhancing therapeutic effect after wounding.

In this aspect, the composition of the present invention may be a food composition.

The food composition of the present invention may be a food composition for enhancing immunity comprising an extract of Aspergillus oryzae as an active ingredient. The food composition may be a functional food composition, specifically a health functional food for improving or preventing immunity.

The food may be a natural product or a processed product containing one or more nutrients, which can be directly eaten through a certain degree of processing. In general terms, the term " health functional food, beverage, food additive, Additives, and the like.

The food of the present invention includes, for example, various foods, beverages, gums, tea, vitamin complex, and health functional food. In addition, in the present invention, the food may contain special nutritional foods (eg, crude oil, infant formula, etc.), meat products, fish products, tofu, jelly, noodles (eg, (Fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kinds of kimchi, pickles, etc.), beverages (such as soy sauce, soybean paste, hot pepper paste, Fermented beverages, two oils, etc.), natural seasoning (e.g., ramen soup, etc.). The food, the health functional food, the beverage, the food additive and the beverage additive can be produced by a usual production method.

In the present invention, the health functional food refers to a food group imparted with added value to function and express the function of the food by physical, biochemical, biotechnological techniques, etc., or to control the bio-defense rhythm of the food composition, Means a food that is designed and manufactured so that the weight control function related to recovery and the like is sufficiently expressed in living bodies.

The health functional food may include food-acceptable food supplementary additives, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of health functional foods.

In the present invention, beverage is a generic term for drinking or enjoying a taste, and is intended to include a health functional beverage. The beverage is not particularly limited as long as it contains the extract as an active ingredient as an essential ingredient at an indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages .

The food composition of the present invention may further comprise a food additive, and the food additive includes all those conforming to the standards and criteria of the relevant item according to the general rules of the Food Additives Ordinance approved by the Korean Food and Drug Administration and the General Test Methods .

The above-mentioned food additives include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as a colorant, licorice extract, crystalline cellulosic, high- - Mixed preparations such as a sodium glutamate preparation, a noodle-added alkaline agent, a preservative preparation, a tar coloring agent and the like.

Examples of the natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose and sucrose and polysaccharides such as dextrin and cyclodextrin, , And sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate may be generally about 1 to 20 g, preferably 5 to 12 g, per 100 ml of the food composition of the present invention. In addition, the composition of the present invention may be used for natural fruit juice, fruit juice drink, And may further contain pulp for the production of beverages.

In addition to the above, the food composition of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such additives is not so critical, but may be selected in the range of 0 to 20,000 parts by weight per 100 parts by weight of the extract of the present invention.

In the present invention, the health functional beverage includes a beverage group to which added value is added to function and express the function of the beverage by using a physical, biochemical, biotechnological technique or the like, a bio-defense rhythm control of the beverage composition, Means a beverage which is processed by being designed so that the body control function related to recovery and the like is sufficiently expressed to a living body.

The health functional beverage is not particularly limited to the ingredients other than the peanut extract of the present invention as an essential ingredient in the indicated ratios and may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient.

Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and xylitol , Sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate may generally be about 1 g to 20 g, preferably 5 g to 12 g per 100 ml of the composition of the present invention.

In addition, the composition of the present invention may further contain flesh for the production of natural fruit juice, fruit juice drink, vegetable drink.

In addition to the above, the food composition of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such additives is not so critical, but may be selected in the range of 0 to 20 parts by weight per 100 parts by weight of the extract of the present invention.

In the present invention, the health functional beverage includes a beverage group to which added value is imparted so that the function of the beverage functions to a specific purpose using physical, biochemical, biotechnological techniques, etc., And restoration, etc., which are designed to be sufficiently expressed in living bodies.

The health functional beverage is not particularly limited to the ingredients other than the peanut extract of the present invention as an essential ingredient in the indicated ratios, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages.

Examples of the natural carbohydrates include polysaccharides such as disaccharides such as monosaccharides such as glucose and fructose, maltose and sucrose, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol . The flavor may be a natural flavor such as tautatin or a stevia extract such as rebaudioside A or glycyrrhizin or a synthetic flavor such as saccharin, aspartame and the like.

The amount of the natural carbohydrate to be added may generally be about 1 to 20 g, preferably 5 to 12 g per 100 ml of the food composition of the present invention. In addition, the composition of the present invention may further contain flesh for the production of natural fruit juice, fruit juice drink, vegetable drink.

In addition, in the food composition for enhancing immunity, which contains the extract of Aspergillus oryzae as an active ingredient, the effective ingredient may be contained in an amount of 0.01 to 15% by weight of the total food, and the beverage composition may contain 0.02 g to 5 g, preferably from 0.3 g to 1 g.

In addition, the present invention provides an animal feed additive for immunity enhancement containing an extract of Aspergillus oryzae as an active ingredient and an animal feed for enhancing the animal feed additive.

The animal feed additive, that is, the composition for animal feed additive, may be in the form of a 20 to 90% high concentrate, powder or granule form.

The composition for animal feed additive of the present invention is a composition containing an organic acid such as citric acid, fumaric acid, adipic acid, lactic acid and malic acid, a phosphate such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymerized phosphate), polyphenol, catechin ), Natural antioxidants such as alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid and phytic acid.

In addition, the animal feed additive and the immunostimulating animal feed comprising the same as the active ingredient include the amino acid, the inorganic salt, the vitamin, the antibiotic substance, the antimicrobial substance, the antioxidant, the antifungal enzyme, the living microorganism Various adjuvants, such as preparations, may be used in the preparation of cereals, such as ground or crushed wheat, oats, barley, corn and rice; Vegetable protein feedstuffs, for example, based on rapeseed, soybeans and sunflower; Animal protein feeds such as blood, meat, bone meal and fish meal; It is preferable to mix all of the dry ingredients comprising the sugar and the milk product such as the various powdered milk and the whey powder and the dry additive and then mix the liquid ingredient with the ingredient to be liquid after heating, In addition to the main components such as fat and vegetable fat, can be used together with materials such as nutritional supplements, digestion and absorption enhancers, growth promoters, disease prevention agents and the like.

The animal feed additive may be administered to an animal alone or in combination with other feed additives in an edible carrier.

The animal feed additives can also be easily administered as top dressing or they can be mixed directly with the animal feed or separately from the feed, in separate oral formulations, by injection, transdermal or in combination with other ingredients. Typically, a single daily dose or a divided daily dose can be used as is well known in the art.

When the animal feed additive is administered separately from animal feed, the dosage form of the composition, as is well known in the art, may be prepared in an immediate release or sustained release formulation in combination with their non-toxic pharmaceutically acceptable excipient . Such edible carriers may be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the dosage form of the extract may be a tablet, capsule, powder, troche or emulsion or top-dressing in finely divided form. When a liquid carrier is used, it may be in the form of a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution. In addition, the dosage form may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution promoting agents and the like.

In addition, the immunostimulating feed comprising the animal feed additive of the present invention as an active ingredient may be any protein-containing organic grain fraction commonly used to meet animal dietary needs. These protein-containing flours usually consist mainly of corn, soy flour or corn / soy flour mix.

The animal feed additive may be added to the animal feed by immersion, spraying or mixing.

The animal scavenging additive can be applied to a number of animal diets including mammals, poultry, and fish. More specifically, the diets may be used in commercial mammals, such as pigs, cows, sheep, goats, laboratory animals (e.g., rats, mice, hamsters and gervilus rats), fur- , And zoo animals (e.g., monkeys and tailless monkeys), as well as livestock (eg, cats and dogs). Typically commercially important poultry may include chickens, turkeys, ducks, geese, pheasants and quails, and may also include commercially reared fish such as trout.

The method of compounding an animal feed comprising an animal feed additive according to the present invention may comprise incorporating the animal feed additive into an animal feed in an amount of about 1 g to 100 g per kg of feed on a dry weight basis.

In addition, after thorough mixing of the feed mixture, lightly viscous granules or granular material may be obtained depending on the degree of crushing of the components, which may be supplied as a mesh or may be formed into a desired discrete shape for further processing and packaging have. At this time, in order to prevent separation during storage, it is preferable to add water to the animal feed, followed by a conventional pelleting, expansion, or extrusion process. Excess water can be dried off.

Particularly, the animal feed additive for immunity enhancement or the feed for immunity enhancement is used for prevention and treatment of diarrhea caused by piglets, respiratory diseases of pigs, diarrhea of cattle, respiratory diseases of chickens, diarrhea and respiratory diseases of dogs and the like .

The form of ingestion of the animal feed additive or immunostimulating feed can be provided in the form of powder or pellet which can be mixed with the feed, or in the form of pellets.

According to another embodiment of the present invention, the extract of Pompeii has an immunostimulating effect. Therefore, the present invention may be a pharmaceutical composition containing Pompeii extract as an active ingredient.

The pharmaceutical composition for enhancing immunity comprising the extract of Aspergillus oryzae as an active ingredient may further comprise, as an active ingredient, a substance having the above-mentioned Aspergillus oryzae extract as an active ingredient or having an immunostimulating effect on the Aspergillus oryzae extract, Depending on the method and intended use, it may further comprise additional components, i.e. pharmaceutically acceptable or nutritionally acceptable carriers, excipients, diluents or subcomponents.

The pharmaceutical compositions for enhancing immunity can be directly applied to animals including humans. The animal is a group of organisms corresponding to plants. It mainly consumes organic matter as nutrients, and differentiates digestive organs, excretory or respiratory organs. Preferably, it is a mammal, more preferably a human.

In detail, when the extract is used as a medicine or used for medicinal or pharmaceutical purposes, the extract can be mixed with a pharmaceutically acceptable carrier or excipient according to a conventional method or diluted with a diluent .

More specifically, the pharmaceutical composition for immunization comprising the extract of Aspergillus oryzae as an active ingredient may further comprise, in addition to the active ingredient, a nutritional supplement, a vitamin, an electrolyte, a flavoring agent, a coloring agent, a thickening agent, a pectic acid and a salt thereof, Organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, and carbonating agents used in carbonated beverages.

The carrier, excipient or diluent may be selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acicia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, 1, selected from the group consisting of water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin, physiological saline, But not limited thereto, and any conventional carrier, excipient or diluent can be used. The components can be added to the active ingredient, i.e., the extract of Chrysanthemum morifolium, either independently or in combination.

The pharmaceutical composition for enhancing immunity comprising the extract of Aspergillus oryzae as an active ingredient may contain a pharmaceutically effective amount of the active ingredient relative to the total weight of the composition, specifically 0.001 to 99.9 wt%, preferably 0.1 wt% To 99% by weight, and more preferably from 1% by weight to 50% by weight. The pharmaceutically effective amount may be appropriately varied depending on the disease and its severity, age, body weight, health condition, sex, administration route and treatment period of the patient.

In addition, the content of the additional component may be added in the range of 0.1 wt% to 20 wt% with respect to the total weight of the pharmaceutical composition for enhancing immunity, but is not limited thereto.

When the pharmaceutical composition for enhancing immunity comprising the extract of Aspergillus oryzae as an active ingredient is weakly formulated, it may be formulated with conventional fillers, extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, And may be used either orally or parenterally.

Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations can be prepared by mixing the above active ingredients, that is, at least one excipient such as starch, calcium carbonate calcium carbonate, sucrose or lactose, and gelatin. 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 and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like May be included.

In addition, the formulation of the pharmaceutical composition for immunostaining comprising the Aspergillus oryzae extract of the present invention as an active ingredient may be in a preferable form depending on the method of use, and it is particularly preferable to provide the rapid, sustained or delayed release of the active ingredient after administration to the mammal By using a method known in the art. Examples of the specific formulations include granules, powders, syrups, liquids, suspensions, tablets, injections, main tablets, cataplasma, capsules, soft or hard gelatin capsules and the like.

Furthermore, the pharmaceutical composition for immunity enhancement comprising the extract of Aspergillus oryzae as an active ingredient of the present invention can be prepared by using a suitable method known in the art or by using the method described in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA) . ≪ / RTI >

The dose of the pharmaceutical composition for enhancing immunity which comprises the extract of Aspergillus oryzae according to the present invention as an active ingredient can be suitably selected by those skilled in the art in consideration of the administration method, the age, sex, and weight of the recipient, . For example, the pharmaceutical composition for enhancing immunity comprising the extract of Aspergillus oryzae as an active ingredient of the present invention may be administered in an amount of from 0.0001 mg / kg to 1000 mg / kg on the basis of the active ingredient, preferably from 0.01 mg / kg to 100 mg / mg / kg < / RTI > The above administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

In addition, the pharmaceutical composition for enhancing immunity comprising the extract of Aspergillus oryzae as an active ingredient of the present invention may further comprise a compound having a known immunity enhancing effect or an extract for a natural product in addition to the above-mentioned effective ingredient, And 5 parts by weight to 200 parts by weight, respectively.

The water extract of Phellodendus majorum, which is an effective ingredient of the immunomodulating composition of the present invention, has been produced by using a long-term pesticide used for medicinal purposes and a safe water solvent. As a result, it has been found that there is no cytotoxicity, Respectively. Specifically, it was confirmed that the level of the cytokine related to the immune response was increased and the number of the infected bacteria was decreased. As a result, it was confirmed that the immunity enhancing effect was very excellent, The composition of the present invention can be widely used in various industrial fields to which an immunity enhancing effect including a food industry, a feed industry and a medicine industry can be applied.

FIG. 1 is a graph showing the cytotoxicity and proliferative effect of the extract of Phellodendus japonica according to an embodiment of the present invention. HAC means a hot water extract of Aralia cordata (HAC) And the vertical axis represents the cell survival rate as compared with the control group.
FIG. 2 is a graph showing the production of intracellular immunocytochemicals by addition of the extract of Phellinus linteus according to an embodiment of the present invention, wherein A is TNF-α, B is IL-1β, C is IL-6 and D is IL -10, and the abscissa of the graph represents the dose of the extract of Phellodendron herb extract and the vertical axis represents the amount of immunocytochemical production.
FIG. 3 is a graph showing mRNA levels of intracellular immunocytokines, that is, intracellular immunocytochemicals before inflammation, by addition of the extract of Phellodendus lobata, according to an embodiment of the present invention, wherein A is TNF-α, B IL-1?, C represents IL-6, and D represents IL-10. The abscissa in the graph represents the dose of the extract of Phellodendron hornblende extract and the vertical axis represents the mRNA production of the immunocytokines.
FIG. 4A is a graph showing the immunity enhancement effect after bacterial infection by addition of the water extract of Phellodendus japonica according to an embodiment of the present invention. FIG. 4A shows changes in body weight of the mice after ingestion of Phellodendus laxifolium extract and infection, 4B shows the number of bacteria in the liver and spleen after listeria infection. The horizontal axis of FIG. 4B indicates the number of bacteria in the experimental group And the vertical axis represents the number of infectious strains (Listeria monocytogenes) per organ weight.
FIG. 5 is a graph showing the immunomodulatory effect by the addition of the extract of Phellodendus javanica according to an embodiment of the present invention. FIG. 5A is a graph showing the amount of immunocytochemical in the liver, and FIG. ), The abscissa of FIGS. 5A and 5B represents the experimental group, and the vertical axis represents the immunocytokine production amount.
FIG. 6 is a photograph showing the antimicrobial activity against Listeria monocytogenes by the flat disc method according to an embodiment of the present invention. On the left side of the third line is the result of treatment with antibiotic Gentamicine as a positive control, and the result of treatment with PBS alone on the right side. On the right side of the second row was the result of treatment with 0.5 mg of the extract of Pomodoropsis lanceolata. The second line on the left side was the result of treatment with 1 mg of Pompeii extract, the second line was the result of 2 mg of Pompeii extract, The results of treating 4 mg of the extract are shown.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

All experimental results were expressed as mean value repeatedly at least three times, and comparison of each group was determined to be significant in the case where each p <0.05 using ANOVA test.

[ Preparation Example . Preparation of papaver extract]

Preparation Example  1. Preparation of extract

Fresh prawns collected from Jeollabuk-do were collected and dried and then ground using a crusher. 50 g of the pulverized pulverized product was mixed with 1 L of water, and extracted for 4 hours using a stirrer under the temperature condition of 120 ° C and the pressure of 2 kg / cm ² , and then the extract was filtered and lyophilized .

The dried Phellodendron hornblende extract was stored at -20 ° C. In the experiment, the Phellodendus hornblende extract was diluted with phosphate buffered saline (PBS), and the concentration of the extract was used for the experiment.

[ Experimental Example  1. Identification of the immune-enhancing effect of the extract of Pompeii]

In order to confirm the immunopotentiating effect of the extract, the bone marrow derived macrophage (BMDM) was used.

First, to isolate bone marrow-derived macrophages, C57B / L6 mice were bred. Specifically, C57B / L6 mice (Sumataka Co., Ltd., Korea) at 8 weeks of age were maintained under temperature conditions of 24 to 26 占 폚, relative humidity of 35 to 45% day), the animals were fed with solid feed and water freely.

The bone marrow-derived macrophages were sacrificed in the breeding experimental animals, the tibia and femur were separated, and the bone marrow was withdrawn by flushing using a syringe. The bone marrow-derived macrophages (BMDM) were separated from the bone marrow using a cell strainer and suspended in DMEM cell culture medium containing fetal calf serum.

First, cytotoxicity (MTT assay) was measured using the separated bone marrow-derived macrophages (BMDM).

Specifically, the cells were injected at a rate of 5 × 10 ⁴ cells per well of a 96-well plate, and cultured for 24 hours at 37.5 ° C. and 5% CO ₂ using an incubator. After that, various concentrations of Phellodendron hirsuta extract (0-500 μg / ml) were added to each well and further cultured for 4 hours. After that, the supernatant of the culture was removed, and MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) reagent was treated and further cultured for 2 hours. Again, the supernatant was removed and the absorbance at 540 nm was measured by treatment with DMSO. The measurement results are shown in Fig.

As shown in Fig. 1, the same number of cells as in the control group treated with no sample was observed after treatment with the extract of Phellodendus japonicus at a maximum concentration of 500 μg / ml, and cytotoxicity was not observed. mu] g / ml compared to the control group not treated at the concentration of [mu] g / ml.

First, the amount of intracellular immunocytokine production and the amount of mRNA of immunocytocytes were measured in relation to the immunoenhancing effect.

Specifically, production of immunocytokines was performed using an enzyme-linked immunosorbent assay (ELISA). During the culturing of the bone marrow-derived macrophages (BMDM), various concentrations of Phellodendron hot-water extracts (0 to 500 μg / ml) were added and cultured for 24 hours at 37.5 ° C. and 5% CO ₂ in an incubator. After the above cultivation, cytokine production was measured at an absorbance of 450 nm using an antibody against each immunocytokine and an ELISA kit (eBioscience, USA). The measurement results are shown in Fig.

As shown in FIG. 2, it was confirmed that the production of cytokine was increased as the concentration of Phellodendron hornblende extract was increased. Specifically, in the case of TNF-α, the production of cytokines from the concentration of 125 μg / , IL-1β and IL-6 significantly increased cytokine production from 500 μg / ml, and IL-10 significantly increased cytokine production from 250 μg / ml. It was confirmed that the production of cine was increased.

In addition, the amount of mRNA of immunocytokines was measured using Real-time PCR.

Specifically, during the culture of the bone marrow-derived macrophages (BMDM), various concentrations of Phellinus linteus extract (0 to 500 μg / ml) were added and cultured for 4 hours at 37.5 ° C. and 5% CO ₂ using an incubator . After incubation, the mRNA content of each immunocytokine was measured using Real-time PCR. The measurement results are shown in Fig.

As shown in FIG. 3, the mRNA content of cytokine was increased as the concentration of the extract of Phellodendron capsicum extract was increased. Specifically, TNF-α (125 μg / ml), IL-1β (62.5 μg / ml) and IL-10 (62.5 μg / ml) were found to increase the mRNA content of cytokines significantly as the concentration increased. In the case of IL-6, the mRNA level of cytokine was the highest at the concentration of 125 μg / ml, and after the concentration, the concentration of IL-6 decreased as the concentration of the IL-6 increased, but the addition of the low concentration (62.5 μg / ml) Still high.

From the results shown in FIG. 2 and FIG. 3, it was confirmed that the intracellular immunocytokine level and the mRNA level of the immunocytokines were increased by the addition of the extract of Phellodendron hornblende extract to enhance the immunity of the body.

In addition, the immune enhancement effect was confirmed by the antimicrobial action in the experimental animal by the addition of the above-mentioned water extract of Phellinus linteus.

Specifically, the experimental animals were raised under the same conditions as above, except that the watery extract of Phellodendus japonicus was taken at a dose of 100 mg / kg per mouse as a test animal for one week. After a week of ingestion, 1 x ^{10 &lt; 9} &gt; CFU of Listeria monocytogenes were infected to the experimental animals by oral administration. In the case of the control group, PBS was administered in place of the hot-water extract.

After oral administration, the body weights of the experimental animals were measured until the 4 days of infection from the day of infection at intervals of 1 day. The results are shown in FIG. 4A. After 96 hours of infection, mice were sacrificed and liver and spleen of the mice were weighed and weighed. The mice were placed in cold PBS containing Triton X-100, metal beads) and Minilys homogenizer (Bertin technologies, France). The homogenized cells were inoculated into BHI agar, cultured at 37 DEG C for 36 hours, and the number of colonies was counted. The results are shown in FIG. 4B.

As shown in FIG. 4A, there was almost no change in the measurement of the weight of the rats when the rats were fed the hot water extract, but it was not significant in the rats fed with PBS, It was confirmed that it recovered again from. Further, as shown in FIG. 4B, the number of bacteria in the liver and spleen was significantly lower in the rats fed the extract of Phellodendus japonica L., It was confirmed that the immune enhancing effect of the Phellodendus japonicus extract was significantly effective also in vivo.

In addition, in order to confirm whether the inhibitory activity of the Listeria monocytogenes was due to an increase in the immune activity in the body or an antimicrobial effect, the extracts of Pseudomonas aeruginosa were treated with 0.25 mg, 0.5 mg, 1 mg, 2 mg and 4 mg, The antimicrobial activity against the bacteria was confirmed.

Specifically, the antimicrobial activity of the Listeria monocytogenes was determined by dissolving the specific amount of the extract in PBS using PBS, and then absorbing the extract solution into a paper disk (Advantec Filter Paper, Japan) having a diameter of 8 mm and a thickness of 1.5 mm After the solvent was completely evaporated, a flat disk on which the solvent was evaporated was placed on the agar plate medium on which the Listeria bacterium was smoothed, cultured for 24 hours, and then the size of the inhibitory circle was determined. Only PBS was used as a negative control, and Gentamicine, an antibiotic, was used as a positive control. The result of checking the size of the inhibition ring is shown in FIG.

As shown in FIG. 6, the antimicrobial activity against Listeria bacterium was measured. As a result, a clear inhibitory effect on the antibiotic Gentamicine was confirmed. However, the inhibitory effect on the antimicrobial extract was not confirmed in all the extracts, It was clearly confirmed that the activity was due to the immune enhancing effect, not the antibacterial activity of the extract itself.

In addition, the amount of inflammatory cytokine production after bacterial infection was measured to further confirm the immune enhancement effect in the body.

Specifically, the immunocytokine content was measured by enzyme-linked immunosorbent assay (ELISA) in the liver and spleen after 48 hours of the infection. The measurement results are shown in Fig.

As shown in FIG. 5A, a high level of cytokine levels was observed in rats fed with pork loin extract, compared to rats fed with PBS as a control. In particular, IL-6, IFN- As shown in FIG. 5B, in the case of cells isolated from the spleen, the difference in the production of immunocytochemicals markedly different from that in almost all cytokines was observed Respectively.

In the present invention, it was confirmed that the level of cytokine involved in the immune response was increased as a result of ingesting the extract of Phellinus linteus for a predetermined period of time using a mouse as a representative experimental animal and examining the immune response. One of the typical infectious bacteria, Listeria monocytogenes ) were infected with rats, it was confirmed that the rats fed the pupae extract significantly decreased the number of bacteria and showed a high level of immunity. From the above results, it has been confirmed that the extract of Phellodendron chinensis has excellent immune enhancing effect in the body.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (8)

A composition for enhancing immunity comprising an extract of Aralia cordata as an active ingredient. delete The method according to claim 1,
Wherein the food composition for immunostaining is a functional food. The method according to claim 1,
Wherein said food composition for immunomodulation is a powder, granule, tablet, capsule or beverage in the form of a formulation. The method according to claim 1,
Wherein the extract is extracted with at least one solvent selected from the group consisting of water and an organic solvent. Aralia cordata ) extract as an active ingredient. The method according to claim 6,
The animal feed additive for immunity enhancement according to any one of claims 1 to 3, An animal feed additive comprising the animal feed additive of claim 6 as an active ingredient.

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