KR20220017738A - Composition for enhancing immunity comprising at least two herbal extracts as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for enhancing immunity and a composition for promoting interferon gamma secretion, comprising at least two herbal extracts as active ingredients. The combination of the at least two extracts synergistically increases the secretion of interferon gamma that promotes immunity in the body without being toxic to cells, so as to be usefully used to enhance immunity.

Description

Composition for enhancing immunity comprising at least two herbal extracts as an active ingredient

The present invention relates to a composition for enhancing immunity comprising two or more kinds of natural herbal extracts as active ingredients. More specifically, the composition of the present invention relates to a health functional food composition and pharmaceutical composition for use in increasing immunity by increasing the secretion of interferon gamma, which enhances immunity in the body.

Immunity is the power that the body kills or neutralizes against an infectious agent such as an external pathogen or virus. Therefore, even if infected with the same external source of infection, people with high immunity do not get sick easily, but people with low immunity are easily infected. Therefore, it can be said that it is most important to increase immunity in order to live a healthy life. Immunity can be divided into innate immunity from birth and acquired immunity acquired through exposure to external infectious agents or vaccination during life.

Immunity began to be known from the fact that those who survived a contagious disease did not subsequently develop the disease. Immunity was first known around 430 BC, and many studies have been conducted since then, and various methods have been devised to increase immunity. One of the most widely used methods is vaccination. Vaccination is a method of enhancing immunity against an infectious agent by artificially injecting a weak infectious agent to stimulate the immune system. Among the methods currently being developed, it is the most effective method for preventing infectious diseases, but since it targets only one disease, the range of action is narrow, and in rare cases, vaccination has limitations that may cause side effects. In fact, every year during the changing seasons, vaccination is carried out to prevent influenza caused by influenza virus, but since the influenza virus that is prevalent every year changes, vaccination is useless if the influenza virus that is prevalent in that year cannot be predicted properly. will arise Therefore, in order to overcome the limitations of such vaccination, it is necessary to broadly increase resistance to various infectious agents through the body's own immunity.

Cytokine is a generic term for proteins secreted by immune cells in the body, and is an important factor in regulating immunity in the body. Among them, interferon is known to mainly play a role in preventing virus proliferation in cells. Interferon was first identified in 1957 as a substance that inhibits the growth of viruses. Interferon produces protein kinase R (PKR) and inhibits intracellular protein synthesis, thereby preventing the protein synthesis necessary for the growth of the virus that infects the cell. do. Interferon is sometimes used to treat MERS or influenza according to this principle, and bats have numerous viruses, but are known to not get sick because they possess a large amount of interferon. In addition to this antiviral role, interferon is widely involved in the activation of immune-related cells such as natural killer cells and macrophages, and is also known to inhibit the growth of cancer cells. Interferon directly recognizes and kills infected cells by activating major histocompatibility complex molecules (MHC) I and II to activate cytotoxic T cells and helper T cells, or activates other immune cells.

In the case of most immune enhancers developed so far, the increase in the number of immune cells is said to be a mechanism to increase immunity. However, even if the number of immune cells increases, if most of the immune cells are inactivated, it cannot be said that immunity is increased, so it is also important to check how activated the immune cells are. Since interferon not only has the ability to increase immunity in the body, but it can also be used as an indicator of various immune cell activities, materials that can increase the concentration of interferon in the body are highly likely to be developed as materials that can increase immunity in the body. can do.

Numerous references are referenced throughout this specification and citations thereof are indicated. The disclosures of the cited documents are incorporated herein by reference in their entirety to more clearly describe the content of the present invention and the level of the art to which the present invention pertains.

Republic of Korea Patent No. 10-1967073

As a result of research efforts to develop a natural product without side effects while exhibiting the effect of enhancing immunity, the present inventors have completed the present invention by revealing that there is a remarkable immune enhancing effect when two or more natural herbal extracts are combined.

Accordingly, an object of the present invention is to provide a health functional food composition for enhancing immunity comprising two or more kinds of herbal extracts as active ingredients.

Another object of the present invention is to provide a pharmaceutical composition for enhancing immunity comprising two or more kinds of herbal extracts as active ingredients.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

One aspect of the present invention is (i) pilbal ( Piper longum ) extract or tangja ( Poncirus trifoliata ) extract; And (ii) red ginseng ( Panax ginseng ) extract or turmeric ( Curcuma longa ) It is to provide a health functional food composition for enhancing immunity, comprising a combination as an active ingredient as an active ingredient. The composition is a leaf mushroom ( Grifola frondosa ) extract and/or cornus oil ( Cornus ) officinalis ) extract may be further included.

Hereinafter, the present invention will be described in detail.

Piper longum is a plant belonging to the genus Pepper and pepper. It smells similar to pepper, but has a slightly bitter taste. The shape is columnar, and countless small granular fruits are attached around the fruit shaft. It is reddish-brown to black-brown. In oriental medicine, it has been used for headache, vomiting, and diarrhea, and the main ingredients are piperlongumine, pipercide, piperine, sylvatine, piperatin, and the like. In the present invention, dried fruits of Piper longum may be used for preparing the extract of pilbal, commercially available ones may be purchased and used, or those harvested or grown in nature may be used.

Poncirus ( Poncirus) trifoliata ) is a deciduous tree belonging to the genus Ruthaceae. It is spherical, and the outer surface is dark brown or brown, and the inner surface is light grayish brown. The immature fruit is mainly used as a medicine, and in oriental medicine, it is mainly used to remove phlegm and stasis, and is also used as an analgesic, antipyretic, and diuretic. The main components are flavonoid glycosides, vitamin C (ascorbic acid), acetic acid, and the like. In the present invention, Poncirus for the preparation of extract The unripe fruits of trifoliata may be used, purchased commercially available, or harvested or grown in nature.

Red ginseng ( Panax ginseng ) is a steamed root of ginseng, a perennial plant belonging to the genus Araliaceae. Elongated, columnar-fusiform, sometimes with a number of superficial roots in the middle. The outer surface is light grayish-brown, and the inner surface is slightly lighter brown. The main ingredient is ginsenoside, and the type varies depending on the location and number of sugars attached, but red ginseng characteristically contains a lot of ginsenoside Rg3. It is used for hangover removal, anti-aging, fatigue recovery, stress relief, immune enhancement, and male infertility treatment. In the present invention, for the production of red ginseng extract, commercially available ones may be purchased and used, or those harvested or grown in nature may be used.

Turmeric ( Curcuma longa ) is a perennial herb belonging to the genus Ginger and Curcuma, and its rhizome is mainly used. It is mainly cultivated in the subtropical region where it rains a lot, and the shape is various, such as oval and spindle-shaped, and there are also bent or branched forms. The main ingredients are curcumin, demethoxycurcumin, zedoarone, turmerone, and eugenol. In oriental medicine, it has been used for the purpose of removing eohyeol and improving digestive function, and has been used for a long time as a medicine and spice, not only in oriental medicine, but also in East and West. For the preparation of turmeric extract in the present invention The rhizome of Curcuma longa can be used, purchased commercially, or harvested or grown in nature.

Mushroom Grifola frondosa belongs to the genus Ginkgo biloba and Mushroom, and it looks like ginkgo leaf-like things are stacked in several layers, so it is called a shiitake mushroom, and there are black, white, and brown colors. Beta-glucan, which is generally contained in mushrooms, is known to be particularly abundant in shiitake mushrooms, so it was approved as an anti-cancer adjuvant by the US FDA in 1998, and clinically it has been reported to have anti-cancer effects. . It is also known as a natural plant with anti-cancer effects, blood sugar suppression, cholesterol lowering, and anti-aging effects. In the present invention, the fruiting body of Grifola frondosa may be used for the preparation of the shiitake mushroom extract, commercially available ones may be purchased and used, or those harvested or grown in nature may be used.

Cornus officinalis ) belongs to the genus Dogwood, and mainly uses its fruits. It is long oval, green at first, then turns dark reddish-purple or purple as it ripens, and has glossy and rough wrinkles. The main ingredients are ursolic acid, oleanolic acid, loganin, quercetin, kaempferol, and organic acids. It is an effective natural plant. In the present invention, the fruit of Cornus officinalis may be used for preparing the cornus oil extract, commercially available ones may be purchased and used, or those harvested or grown in nature may be used.

The active ingredient included in the composition for enhancing immunity of the present invention may have a remarkable synergistic effect in relation to immunity enhancement compared to each single substance by combining two or more kinds of the herbal extracts. The combination of two or more of the herbal extracts is preferably (i) Piper longum extract or Poncirus trifoliata ) extract; And (ii) red ginseng ( Panax ginseng ) extract or turmeric ( Curcuma ) longa ) extracts.

As used herein, the term “improving immunity” refers to any phenomenon in which a factor that increases immunity in the body is increased or a factor that suppresses immunity in the body is decreased by administration of the composition.

The extract of the natural herbal ingredient of the present invention may include a simple water extract, an organic solvent extract, and a solvent fraction and a polysaccharide fraction of the extract, and may be a mixture or complex of the simple water extract, organic solvent extract, solvent fraction and polysaccharide fraction. can

As used herein, the term "extract" may be obtained by extraction with one or more solvents selected from the group consisting of water, C1 to C4 lower alcohols, 1,3-butylene glycol, and ethyl acetate, but is limited thereto. doesn't happen Specifically, the preparation process of the extract of the present invention may largely include an extraction step, a concentration step, and a drying step. In the manufacturing process of the extract, pilbal, tangja, red ginseng, turmeric, shiitake mushroom or cornus oil can be used without limitation, such as cultivated or commercially available ones.

In the extraction step of the present invention, the extraction solvent may be obtained by extraction with one or more solvents selected from the group consisting of water, C1 to C4 lower alcohols, 1,3-butylene glycol, and ethyl acetate, preferably may be fermented alcohol, but is not limited thereto. The extraction method may use an extraction method such as fermented alcohol extraction, immersion extraction, reflux cooling extraction, or ultrasonic extraction, but is not limited thereto. The extraction solvent may be extracted by adding 1 to 10 times the amount of dried pilbal, tangja, red ginseng, turmeric, shiitake mushrooms or cornus oil, and the extraction temperature may be 20° C. to 100° C., specifically 50° C. to 80° C. It may be ℃, but is not limited thereto. In addition, the extraction time may be 1 hour to 48 hours, specifically may be 2 hours to 8 hours, but is not limited thereto. In addition, the number of times of extraction may be 1 to 5 times, and specifically, extraction may be repeated 1 to 3 times, but is not limited thereto.

In the concentration step of the present invention, a process of concentrating the extract under reduced pressure may be further included, and the vacuum concentration may be performed using a vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto.

The drying step of the present invention may further include drying the extract that has undergone the concentration under reduced pressure, and the drying may be vacuum drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto. .

The extract of the present invention may enhance human immunity by increasing the secretion of interferon gamma through activation of immune cells.

The composition comprising two or more kinds of extracts of the present invention described above as an active ingredient can be used as a health functional food composition for enhancing immunity.

As used herein, the term "food" includes, for example, various foods, beverages, gum, tea, vitamin complexes, health functional foods, and the like, and includes all foods in a conventional sense. The food can be prepared in dosage forms such as tablets, granules, powders, capsules, liquid solutions and pills according to a known manufacturing method, and the content of the extract according to the present invention is 0.0001 based on the total weight of the food composition depending on the dosage form. It can be adjusted to 100% by weight to 100% by weight, but is not limited thereto. There is no particular limitation on other ingredients except for including the extract according to the present invention as an active ingredient, and various conventional flavoring agents or natural carbohydrates may be included as additional ingredients.

Examples of the natural carbohydrate include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate may be generally about 1 g to 20 g, specifically, about 5 g to 12 g per 100 ml of the composition of the present invention.

In the present invention, the term "health functional (sex) food" is the same term as food for special health use (FoSHU), and in addition to nutritional supply, it has medical and medical effects processed to efficiently exhibit bioregulatory functions. It means high food. Here, "function (sex)" means to obtain a useful effect for health purposes such as regulating nutrients or physiological action with respect to the structure and function of the human body. The food of the present invention can be prepared by a method commonly used in the art, and during the manufacture, it can be prepared by adding raw materials and components commonly added in the art. In addition, the formulation of the food may be prepared without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be prepared in various types of dosage forms, and unlike general drugs, there is no side effect that may occur during long-term administration of the drug using food as a raw material, and has excellent portability.

Specifically, the health functional food is a food prepared by adding the composition of the present invention to food materials such as beverages, teas, spices, gum, and confectionery, or encapsulating, powdering, suspension, etc., and when ingested, It means to bring an effect, but unlike general drugs, it has the advantage that there are no side effects that may occur when taking the drug for a long time using food as a raw material.

The food composition may further include a physiologically acceptable carrier, the type of carrier is not particularly limited and any carrier commonly used in the art may be used.

In addition, the food composition may include additional ingredients that are commonly used in food compositions to improve odor, taste, vision, and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, and the like may be included. In addition, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), chromium (Cr); and amino acids such as lysine, tryptophan, cysteine, and valine.

In addition, the food composition includes a preservative (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), a disinfectant (bleaching powder and high bleaching powder, sodium hypochlorite, etc.), an antioxidant (butylhydroxyanisole (BHA), butyl hydro Loxytoluene (BHT), etc.), coloring agents (tar pigments, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasonings (MSG sodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, etc.) , sodium, etc.), flavorings (vanillin, lactones, etc.), swelling agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (foaming agents), film agents, gum base agents, foam inhibitors, solvents, improving agents, etc. It may contain food additives. The additive may be selected according to the type of food and used in an appropriate amount.

As an example of the food composition of the present invention, it may be used as a health drink composition, and in this case, it may contain various flavoring agents or natural carbohydrates as an additional component like a conventional drink. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as taumatine, stevia extract; A synthetic sweetener such as saccharin or aspartame may be used. The ratio of the natural carbohydrate may be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g per 100 ml of the health beverage composition of the present invention.

In addition to the above, the health drink composition includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol, a carbonation agent, or the like. In addition, it may contain the pulp for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. The proportion of the components may be selected from 0 parts by weight to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

All ingredients described in the present invention, preferably, relevant laws and regulations, such as Korea, China, the United States, Europe, Japan, etc. (for example, the Food Standards Code (Korea), Food Additives Code (Korea), Health Functional Foods Code (Korea) Do not exceed the maximum use value stipulated in Korea)), etc. That is, preferably, the food or food composition according to the present invention contains the ingredients according to the present invention within the content limit allowed by the relevant laws and regulations of each country.

In addition, the composition comprising two or more extracts of the present invention as an active ingredient may be used as a pharmaceutical composition for enhancing immunity.

As used herein, the term "pharmaceutical composition" refers to one prepared for the purpose of enhancing immunity, and each may be formulated in various forms according to a conventional method and used.

The pharmaceutical composition of the present invention may include one or more active ingredients exhibiting the same or similar function in addition to the above ingredients. The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive includes starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate. , lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, Calcium stearate, sucrose, dextrose, sorbitol and talc and the like may be used. The pharmaceutically acceptable additive according to the present invention may be included in an amount of 0.1 to 90 parts by weight based on the composition, but is not limited thereto.

The pharmaceutical composition of the present invention may be administered in various oral and parenteral formulations during actual clinical administration, and when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents or excipients such as surfactants It can be prepared using Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract of the present invention, for example, starch, calcium carbonate, sucrose (Sucrose), may be prepared by mixing lactose (Lactose) or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Liquid formulations for oral use include suspensions, solutions, emulsions, and syrups, and various excipients such as wetting agents, sweetening agents, fragrances, and preservatives in addition to commonly used simple diluents such as water and liquid paraffin may be included. . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

The pharmaceutical composition of the present invention may be administered orally or administered parenterally according to a desired method, and when administered parenterally, external skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection You can choose the injection method. The dosage may vary depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and severity of disease.

The dosage of the pharmaceutical composition of the present invention varies depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate and severity of disease, and the daily dosage is the dosage of the present invention. It may be 0.0001 mg/kg to 100 mg/kg based on the amount of the extract, specifically 0.001 mg/kg to 10 mg/kg, and may be administered once to 6 times a day, but is not limited thereto.

In addition, the composition comprising two or more extracts of the present invention as an active ingredient may be used as a quasi-drug composition for enhancing immunity.

The quasi-drug composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent if necessary in addition to the above components. The pharmaceutically acceptable carrier, excipient or diluent is not limited as long as it does not impair the effects of the present invention, and includes, for example, fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc. may include

The quasi-drug composition of the present invention may include, but is not limited to, disinfectant cleaner, shower foam, ointment, wet tissue, coating agent, etc. may be appropriately selected from among the techniques of

It was confirmed that the mixture containing two or more kinds of herbal extracts of the present invention as an active ingredient synergistically enhances interferon gamma when treated with a mixture rather than when each extract is treated alone (Table 1 - Table 8). Since interferon gamma plays an important role in immunity in the body, this composition can be used for enhancing immunity in the body.

Therefore, it was confirmed that the composition comprising two or more kinds of herbal extracts as an active ingredient can be used as a health functional food composition for enhancing immunity, a pharmaceutical composition for enhancing immunity, or a quasi-drug composition for enhancing immunity.

Since the composition comprising two or more kinds of herbal extracts of the present invention as an active ingredient promotes the secretion of interferon gamma without being toxic to cells, it can be usefully used to enhance immunity.

Hereinafter, the present invention will be described in detail by the following Examples and Preparation Examples. However, the following Examples and Preparation Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the Examples and Preparation Examples. In addition, since these examples are only for the purpose of helping the understanding of the present invention, the scope of the present invention is not limited thereto in any sense.

Example

I. Preparation of extract

production example One: leaf mushroom , curcuma , cornflower oil, tangja, red ginseng or must-have Preparation of extracts

Leaf mushroom, turmeric, cornus oil, tangja, red ginseng or pilbal extract was prepared by a manufacturing method comprising the following steps.

1) Washing and drying shiitake mushrooms, turmeric, cornflower oil, tangja, red ginseng or pilbals, then pulverizing them into powder;

2) adding fermented alcohol to 1000 kg of shiitake mushroom, turmeric, cornflower oil, tangja, red ginseng or pilbal powder obtained in step 1) and extracting at 60° C. for 2 hours;

3) filtering the extract of step 2);

4) concentrating the filtered extract of step 3);

5) Drying the concentrated extract of step 4) to obtain about 1000 g of extract of shiitake mushroom, turmeric, cornflower oil, tangja, red ginseng or pilbal.

production example 2: leaf mushroom , curcuma , cornflower oil, tangja, red ginseng or must-have Preparation of extracts

Leaf mushroom, turmeric, cornus oil, tangja, red ginseng or pilbal extract was prepared by a manufacturing method comprising the following steps.

1) Washing and drying shiitake mushrooms, turmeric, cornflower oil, tangja, red ginseng or pilbals, then pulverizing them into powder;

2) extracting at room temperature for 3 days by adding water to 1000 kg of shiitake mushroom, turmeric, cornflower oil, tangja, red ginseng or pilbal powder obtained in step 1);

3) filtering the extract of step 2);

4) concentrating the filtered extract of step 3);

5) Drying the concentrated extract of step 4) to obtain about 1000 g of extract of shiitake mushroom, turmeric, cornflower oil, tangja, red ginseng or pilbal.

production example 3: leaf mushroom , curcuma , cornflower oil, tangja, red ginseng or must-have Preparation of extracts

Leaf mushroom, turmeric, cornus oil, tangja, red ginseng or pilbal extract was prepared by a manufacturing method comprising the following steps.

1) Washing and drying shiitake mushrooms, turmeric, cornflower oil, tangja, red ginseng or pilbals, then pulverizing them into powder;

2) adding water to 1000 kg of shiitake mushroom, turmeric, cornus oil, tangja, red ginseng or pilbal powder obtained in step 1) and extracting it at 120° C. for 1 hour;

3) filtering the extract of step 2);

4) concentrating the filtered extract of step 3);

5) Drying the concentrated extract of step 4) to obtain about 1000 g of extract of shiitake mushroom, turmeric, cornflower oil, tangja, red ginseng or pilbal.

II. Immune boosting effect evaluation

Example One: leaf mushroom extract and curcuma Interferon-gamma secretion promoting effect of the extract mixture

Mushrooms were extracted with water at room temperature for 2 days according to Preparation Example 2 using the mycelium, and turmeric was extracted with rhizomes using rhizomes at 60°C and 70% fermented alcohol for 2 hours according to Preparation Example 1.

Interferon gamma (IFNγ) is one of the representative cytokines involved in the immune system in the body, and has been reported to play an important role in regulating the immune response to viruses, harmful bacteria, fungi, etc. that cause diseases in the human body. Therefore, when the secretion of interferon gamma increases, the body's immunity increases and the growth of viruses, harmful bacteria, and fungi that cause disease in the human body is inhibited or killed, making it difficult to get sick.

Cells mainly involved in the production and secretion of interferon gamma in the body include natural killer cells (NK cells) that produce interferon gamma, and several types of cytokines that make natural killer cells produce interferon gamma. There are secreting macrophages. In the body, the two interact and secrete interferon gamma to regulate the body's immunity.

Therefore, the present inventors treated macrophages, which are one of the cells mainly involved in the production and secretion of interferon gamma in the body, with a mixture of shiitake mushroom extract and turmeric extract at various concentrations, and then the macrophages were treated with a mixture of shiitake mushroom extract and turmeric extract After processing the macrophage culture medium containing cytokines secreted in response to natural killer cells, another cell mainly involved in the production and secretion of interferon gamma in the body, the concentration of interferon gamma secreted from the natural killer cells is enzymatically combined. The immunoprecipitation assay (Enzyme-linked Immunosorbent Assay, ELISA) was analyzed as follows.

The day before the experiment to analyze the interferon gamma concentration, the interferon gamma capture antibody was diluted to 2 μg/ml, 100 μl per well was placed in an immunoplate, and placed at room temperature. The next day, all the capture antibody was removed, washed 3 times with 1X wash buffer, and 300ul of reagent diluent 2 diluted with 1X was added to each well to block all portions to which the capture antibody was not attached. During blocking, the culture medium of natural killer cells to be analyzed for interferon gamma concentration was centrifuged and the supernatant was taken. One hour after the start of blocking, all reagent diluent 2 was removed, washed 3 times with 1X wash buffer, and then 100 ul of the supernatant taken above was added per well and mixed at 300 rpm. At this time, 9.375 ng/ml, 18.75 ng/ml, 37.5 ng/ml, 75 ng/ml, 150 ng/ml, 300 ng/ml, 600 ng of the interferon gamma standard solution was added to obtain the interferon gamma concentration-absorbance standard curve. It was diluted to /ml and put into the well next to the well containing the natural killer cell culture solution, 100 ul per well. After 2 hours, both the natural killer cell culture medium and the interferon gamma standard solution were removed, washed 3 times with 1X wash buffer, and 100 μl of the detection antibody diluted to 0.125 μg/ml was added per well and shaken at 300 rpm. After 2 hours, all the detection antibodies were removed, washed 3 times with 1X wash buffer, and 100 ul of streptoavidin-HRP diluted with 1X was added per well, shielded from light, and then shaken at 300 rpm. After 20 minutes, all streptoavidin-HRP was removed, 100ul of the substrate was added per well, and the light was blocked, followed by shaking at 300rpm for 5 minutes. After shaking for 5 minutes, it was confirmed that the color of the wells into which the interferon gamma standard solution was added changed to blue in a concentration-dependent manner. Immediately after the addition of 2N sulfuric acid, the green color changed to yellow while mixing well. After measuring the absorbance at 450 nm and 570 nm, the absorbance of each well was substituted into the interferon gamma concentration-absorbance standard curve obtained from the standard solution to calculate each interferon gamma concentration, and the negative control was converted to 1 in Table 1 below. It is written as one fold value.

Mushroom extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Turmeric Extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.00 4.22 6.08 8.19 11.25 21.38 38.60 34.44 30.26 24.25 20.92 6.44

As a result, it was confirmed that the increase in interferon gamma concentration increased synergistically when the shiitake mushroom extract and the turmeric extract were mixed and treated to 100 μg/ml, rather than when the shiitake mushroom extract and the turmeric extract were treated alone at 100 μg/ml. became In addition, the increase in the interferon gamma concentration was the largest when the shiitake mushroom extract and the turmeric extract were mixed and treated at a ratio of 50:50. As the ratio of the extract increased, the increase in the interferon gamma concentration rapidly decreased (Table 1).

Therefore, it can be seen that the shiitake mushroom extract and the turmeric extract synergistically increase the interferon gamma concentration, so that there is a synergistic immune enhancing effect in the body.

Example 2: leaf mushroom Interferon-gamma secretion promoting effect of extract and cornus extract mixture

The same extract as prepared in Example 1 was used for the extract of shiitake mushroom, and the cornus extract was extracted with water at 100° C. for 2 hours according to Preparation Example 3 using the fruit.

The present inventors treated macrophages, which are one of the cells mainly involved in the production and secretion of interferon gamma in the body, in various concentrations by the above method, and then treated the mixture of the shiitake mushroom extract and cornus extract at various concentrations. The concentration of interferon gamma secreted from natural killer cells after the macrophage culture medium containing cytokines secreted in response to the mixture of extracts is treated with natural killer cells, another cell mainly involved in the production and secretion of interferon gamma in the body. was analyzed by enzyme-linked immunoprecipitation assay (ELISA).

The interferon gamma concentration was calculated in the culture medium obtained by culturing macrophages and natural killer cells after treating the mixture of shiitake mushroom extract and cornus extract in the same way as above. described.

Mushroom extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Cornus extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.00 4.92 7.83 12.89 16.00 19.67 24.17 19.42 15.06 11.25 5.50 2.61

As a result, the increase in the concentration of interferon gamma was synergistically greater when the shiitake mushroom extract and the Cornus officinalis extract were treated to 100 µg/ml by mixing, rather than 100 µg/ml alone. In addition, the increase in the interferon gamma concentration was the largest when the shiitake mushroom extract and the cornus mushroom extract were mixed and treated at a ratio of 50:50. decreased (Table 2).

Therefore, it can be seen that the leaf mushroom extract and the cornflower extract synergistically increase the interferon gamma concentration, so that there is a synergistic immune enhancement effect in the body.

Example 3: leaf mushroom Efficacy of interferon gamma secretion promoting effect of extract and tangja extract mixture

As for the leaf mushroom extract, the same one prepared in Example 1 was used, and Taengja was extracted from the fruit with 70% fermented alcohol at 60° C. for 2 hours according to Preparation Example 1.

The present inventors treated macrophages, which are one of the cells mainly involved in the generation and secretion of interferon gamma in the body, with various concentrations of a mixture of shiitake mushroom extract and tangja extract in the above method, and then the macrophages The concentration of interferon gamma secreted from natural killer cells after the macrophage culture medium containing cytokines secreted in response to the mixture of extracts is treated with natural killer cells, another cell mainly involved in the production and secretion of interferon gamma in the body. was analyzed by enzyme-linked immunoprecipitation assay (ELISA).

The interferon gamma concentration was calculated in the culture medium obtained by culturing the mixture of shiitake mushroom extract and tangja extract in the same way as macrophages and natural killer cells. described.

Mushroom extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Tangja extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.00 4.22 5.86 9.91 14.66 21.49 31.85 26.08 23.43 19.61 15.34 5.86

As a result, the increase in the concentration of interferon gamma was synergistically greater when treated to 100 μg/ml by mixing the shiitake mushroom extract and dandelion extract than when the shiitake mushroom extract and Taengja extract alone were treated at 100 μg/ml. In addition, the increase in interferon gamma concentration was the largest when the extracts of shiitake mushroom and taengjae extract were mixed and treated at a ratio of 50:50. As the ratio of the extract increased, the increase in the interferon gamma concentration decreased sharply (Table 3).

Therefore, it was found that the shiitake mushroom extract and Taengja extract synergistically increase the interferon gamma concentration, and thus have a synergistic immune-enhancing effect in the body.

Example 4: leaf mushroom Interferon-gamma secretion promoting effect of extract and red ginseng extract mixture

The leaf mushroom extract was the same as that prepared in Example 1, and the red ginseng was extracted using rhizomes and 70% fermented alcohol at 60°C according to Preparation Example 1 for 2 hours.

The present inventors treated macrophages, which are one of the cells mainly involved in the generation and secretion of interferon gamma in the body, with various concentrations of a mixture of shiitake mushroom extract and red ginseng extract in the above method, and then The concentration of interferon gamma secreted from natural killer cells after the macrophage culture medium containing cytokines secreted in response to the mixture of extracts is treated with natural killer cells, another cell mainly involved in the production and secretion of interferon gamma in the body. was analyzed by enzyme-linked immunoprecipitation assay (ELISA).

The interferon gamma concentration was calculated in the culture medium obtained by culturing after treating the mixture of leaf mushroom extract and red ginseng extract with macrophages and natural killer cells in the same way as above. described.

Mushroom extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Red Ginseng Extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.00 4.92 5.94 7.81 11.08 13.31 21.36 15.39 13.00 7.69 3.89 2.03

As a result, the increase in the interferon gamma concentration was synergistically greater when the shiitake mushroom extract and the red ginseng extract were treated to be 100 μg/ml by mixing the shiitake mushroom extract and the red ginseng extract than when the ginseng extract and the red ginseng extract were treated alone. In addition, the increase in the interferon gamma concentration was the largest when the shiitake mushroom extract and the red ginseng extract were mixed and treated at a ratio of 50:50. decreased (Table 4).

Therefore, it was found that the ginseng extract and the red ginseng extract synergistically increase the interferon gamma concentration, and thus have a synergistic immune-enhancing effect in the body.

Example 5: Red ginseng extract and must-have Interferon-gamma secretion promoting effect of the extract mixture

Red ginseng was extracted using rhizomes using 70% fermented alcohol at 60° C. for 2 hours according to Preparation Example 1, and pilbals were crushed and extracted with DMSO, followed by filtration.

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by culturing macrophages and natural killer cells after treating the mixture of red ginseng extract and pilbal extract in the same manner as above, and in Table 5 below, the negative control group was converted into a fold It was written as a value.

Red Ginseng Extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Pilbal Extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.0 3.4 8.0 7.5 5.3 5.5 5.6 7.0 6.0 6.8 11.3 11.8

As a result of the above experiment, since the interferon gamma concentration was increased when the red ginseng extract was added to the red ginseng extract compared to the case of red ginseng alone, it was confirmed that the immunity enhancing effect of red ginseng was enhanced by the peel.

Example 6: Interferon gamma secretion promoting effect of red ginseng extract and tangja extract mixture

Red ginseng was extracted using rhizomes with 70% fermented alcohol at 60°C according to Preparation Example 1 for 2 hours, and Taengja was extracted using fruits with 70% fermented alcohol at 60°C according to Preparation Example 1 for 2 hours.

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by culturing macrophages and natural killer cells after treating the mixture of red ginseng extract and tangja extract in the same manner as above, and in Table 6 below, the negative control group was converted into a fold It was written as a value.

Red Ginseng Extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Tangja Extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.0 3.4 6.1 9.1 7.7 9.9 10.8 13.9 15.1 13.5 15.0 13.3

As a result of the above experiment, since the interferon gamma concentration was increased when the tangja extract was added to the red ginseng extract compared to when the red ginseng was treated alone, it was confirmed that the immunity enhancing effect of the red ginseng was enhanced by the tangja.

Example 7: curcuma extract and must-have Interferon-gamma secretion promoting effect of the extract mixture

As the turmeric extract, the same one prepared in Example 1 was used, and the pilbal was filtered after crushing the fruit and extracting it with DMSO.

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by culturing macrophages and natural killer cells after treating the mixture of turmeric extract and pilbal extract in the same way as above, and in Table 7 below, the negative control group was converted into a fold It was written as a value.

Turmeric Extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Pilbal Extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.0 6.7 12.1 16.1 16.2 16.3 18.9 21.3 19.4 18.3 18.4 11.8

As a result of the above experiment, since the turmeric extract and the pilbal extract synergistically increased the interferon gamma concentration, it was found that there was a synergistic immune enhancing effect in the body.

Example 8: curcuma Efficacy of interferon gamma secretion promoting effect of extract and tangja extract mixture

The turmeric extract was the same as that prepared in Example 1, and tangja was extracted from the fruit with 70% fermented alcohol at 60°C for 2 hours according to Preparation Example 1.

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by culturing macrophages and natural killer cells after treating the mixture of turmeric extract and tangja extract in the same way as above, and in Table 8 below, the negative control group was converted into a fold It was written as a value.

Turmeric Extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Tangja Extract (ppm) 0 0 10 20 30 40 50 60 70 80 90 100 IFNγ concentration (fold) 1.0 6.7 12.5 13.0 14.5 18.2 16.7 15.1 13.6 12.7 12.2 13.3

As a result of the above experiment, when the turmeric extract was added to the tangja extract, the interferon gamma concentration was increased as compared to the turmeric alone treatment, so it was confirmed that the immunity enhancing effect of turmeric was enhanced by the turmeric.

III. Food sensory evaluation

As shown in Table 9 below, a beverage was prepared and the sensory was evaluated after drinking it to 50 healthy men and women aged 18 to 60 years.

Sugar 5% citric acid 0.2% Spices 0.1% effective ingredients Amounts listed in Tables 10 to 13 below water Remaining amount (up to 100%)

Sensory Evaluation Institute installed individual booths with partitions so that they could concentrate without exchanging opinions with each other, and a sink for mouth washing was installed in the individual booths. The temperature was maintained at 20~25℃ and humidity at 50~60%, and ventilation facilities were installed to easily remove odors. The drinking results were evaluated by the following sensory evaluation scale, and the average value was calculated.

<sensory evaluation scale>

Strong (5), Tasty (4), Normal (3), Mild (2), Unpalatable (1)

The results of beverages prepared by mixing the pilbal extract obtained according to the above scale with red ginseng or turmeric extract are shown in Tables 10 and 11, respectively.

Red Ginseng (ppm) 100 60 50 40 0 Feet (ppm) 0 40 50 60 100 spicy 1.4 1.4 1.4 1.6 4.8 bitter 2.6 1.8 1.6 1.8 4.6 Sour taste 3.2 2.0 1.8 2.0 2.4

Turmeric (ppm) 100 60 50 40 0 Feet (ppm) 0 40 50 60 100 spicy 3.6 1.4 1.6 1.8 4.8 bitter 3.2 1.2 1.2 1.8 4.6 Sour taste 3.8 1.4 1.2 1.0 2.4

As a result of the experiment, as can be seen in Tables 10 and 11, the pilbal extract alone had strong pungent, bitter and sour taste, resulting in low palatability as a food. In addition, it was confirmed that the bitter and sour taste was strong even with the red ginseng extract alone, and the spicy, bitter and sour taste was strong even with the turmeric extract alone. However, when the pilbal extract was mixed with the red ginseng or turmeric extract, the loss of taste was confirmed in the rather spicy, bitter and sour taste, and the taste as a food was greatly improved.

In addition, according to the above scale, the results of beverages prepared by mixing tangja extract with red ginseng or turmeric extract are shown in Tables 12 and 13 below, respectively.

Red Ginseng (ppm) 100 60 50 40 0 Tank (ppm) 0 40 50 60 100 spicy 1.4 1.0 0.8 0.8 1.2 bitter 2.6 1.2 1.4 1.2 4.4 Sour taste 3.2 1.4 1.0 1.2 4.8

Turmeric (ppm) 100 60 50 40 0 Tank (ppm) 0 40 50 60 100 spicy 3.6 1.0 0.8 0.8 1.2 bitter 3.2 1.8 1.4 1.4 4.4 Sour taste 3.8 2.0 1.8 1.8 4.8

As a result of the experiment, as can be seen in Tables 12 and 13 above, the bitterness and sourness of the extract alone were particularly strong, and thus the palatability as a food was low. In addition, it was confirmed that the bitter and sour taste was strong even with the red ginseng extract alone and the turmeric extract alone. However, when the tangja extract was mixed with red ginseng or turmeric extract, rather bitter and sour taste, loss of taste was confirmed, and the taste as food was greatly improved.

In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims to be described later and their equivalents.

Claims (5)

(i) Piper longum extract or Poncirus trifoliata ) extract; and
(ii) red ginseng ( Panax ginseng ) extract or turmeric ( Curcuma ) longa ) extract
A health functional food composition for enhancing immunity, comprising a combination of as an active ingredient. The health functional food according to claim 1, wherein the extract is extracted with a solvent selected from the group consisting of water, C1 to C4 lower alcohol, 1,3-butylene glycol, ethyl acetate, and a mixed solvent thereof. composition. The health functional food composition according to claim 2, wherein the lower alcohol is methanol, ethanol, propanol or butanol. The health functional food composition according to claim 1, wherein the active ingredient promotes the secretion of interferon gamma. A food comprising the health functional food composition according to any one of claims 1 to 4.