KR102577328B1 - 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 two or more herbal extracts as active ingredients. The combination of the above two or more extracts synergistically increases the secretion of interferon gamma, which promotes immunity in the body without being toxic to cells, and can 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 containing two or more natural herbal extracts as active ingredients. More specifically, the composition of the present invention relates to a health functional food composition and a pharmaceutical composition for the purpose of increasing immunity by increasing the secretion of interferon gamma, which increases immunity in the body.

Immunity is the power of the human body to fight against infectious agents such as external pathogens or viruses and kill or neutralize them. Therefore, even if infected with the same external infectious agent, people with high immunity do not get sick, but people with low immunity are easily infected. Therefore, in order to live a healthy life, it is most important to increase immunity. Immunity can be divided into innate immunity, which a person has from birth, and acquired immunity, which is acquired during life through exposure to external infectious agents or vaccination.

Immunity began to be known from the fact that people who survived an infectious disease did not develop the disease later. Immunity was first known around 430 BC, and since then, much research has been conducted and various methods to increase immunity have been devised. Among these, one of the most widely used methods is vaccination. Vaccination is a method of strengthening immunity against an infectious agent by artificially introducing a weak infectious agent and stimulating the immune system. Among the currently developed methods, it is the most efficient method to prevent infectious diseases, but it has a narrow scope of action because it targets only one disease, and in rare cases, vaccination has the limitation of causing side effects. In fact, at the change of seasons every year, vaccination is administered to prevent flu caused by influenza viruses, but since the influenza viruses prevalent each year are different, vaccination is useless if the influenza viruses prevalent that year cannot be properly predicted. comes into existence. Therefore, in order to overcome these limitations of vaccination, resistance to various infectious agents must be increased through the body's own immunity.

Cytokine is a general term for proteins secreted by immune cells in the body, and is an important element that regulates the body's immunity. Among them, interferon is known to mainly play a role in preventing viruses from multiplying within cells. Interferon was first discovered in 1957 as a substance that inhibits the growth of viruses. Interferon prevents protein synthesis necessary for the growth of viruses that infect cells by producing protein kinase R (PKR) and inhibiting intracellular protein synthesis. It also inhibits virus growth by destroying viral RNA by producing RNAse L. do. Based on this principle, interferon is sometimes used to treat MERS or pandemic influenza, and although bats possess numerous viruses, they are known to possess a large amount of interferon and thus do not get sick. In addition to its 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 activates major histocompatibility complex molecules (MHC) I and II to activate cytotoxic T cells and helper T cells, thereby directly recognizing and killing infected cells, or activating other immune cells.

In the case of most immune enhancing agents developed to date, the mechanism for increasing immunity is an increase in the number of immune cells. However, even if the number of immune cells increases, if most immune cells are inactivated, immunity cannot be said to have increased, so it is also important to check how activated the immune cells are. Since interferon not only has the activity of increasing the body's immunity on its own, but can also be used as an indicator of the activity of various immune cells, there is a high possibility that materials that can increase the concentration of interferon in the body can be developed into materials that can increase the body's immunity. can do.

Numerous documents are referenced and cited throughout this specification. The disclosures of the cited documents are incorporated herein by reference in their entirety to more clearly explain the content of the present invention and the level of technical field to which the present invention pertains.

Republic of Korea Patent No. 10-1967073

As a result of research efforts to develop a natural product that exhibits an immunity-enhancing effect and has no side effects, the present inventors have completed the present invention by discovering that combining two or more natural herbal extracts has a significant immunity-enhancing effect.

Therefore, the purpose of the present invention is to provide a health functional food composition for improving immunity containing two or more herbal extracts as active ingredients.

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

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

One aspect of the present invention is (i) Piper longum extract or Poncirus trifoliata extract; and (ii) a combination of red ginseng extract or turmeric ( Curcuma longa ) extract as an active ingredient, to provide a health functional food composition for improving immunity. The composition may further include maitake mushroom ( Grifola frondosa ) extract and/or Cornus officinalis extract.

Hereinafter, the present invention will be described in detail.

Piper longum is a plant belonging to the pepper genus of the pepper family. It smells similar to pepper, but tastes a little more bitter. The shape is cylindrical, and numerous small grain-shaped fruits are attached around the fruit axis. It is reddish brown to black brown in color. In oriental medicine, it is used for headaches, vomiting, and diarrhea, and its main ingredients include piperlongumine, pipercide, piperine, sylvatine, and piplartine. In the present invention, the dried fruit of Piper longum can be used to prepare the extract, commercially available ones can be purchased and used, or those collected or cultivated in nature can be used.

Tangja ( Poncirus) trifoliata ) is a deciduous tree belonging to the Rutaceae genus. It is spherical, and the outer surface is dark brown to brown, and the inner surface is light grey-brown. The immature fruit is mainly used as a medicine. In oriental medicine, it is mainly used to remove phlegm and stasis, and is also used as a pain reliever, fever reducer, and diuretic. The main ingredients are flavonoid glycosides, vitamin C (ascorbic acid), and acetic acid. For the production of Tangja extract in the present invention , Poncirus You can use the unripe fruits of trifoliata , purchase them commercially, or use those collected or grown in nature.

Red ginseng is the steamed root of ginseng, a perennial plant belonging to the ginseng genus of the Araliaceae family. It has a long, thin columnar-spindle shape, and sometimes has multiple outer roots growing from the middle. The outer surface is light gray-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 the purposes of eliminating hangovers, preventing aging, recovering from fatigue, relieving stress, enhancing immunity, and treating male infertility. In order to produce red ginseng extract in the present invention, commercially available ginseng extract can be purchased and used, or one collected or cultivated in nature can be used.

Turmeric ( Curcuma longa ) is a perennial plant of the ginger and curcuma genus, and its rhizome is mainly used. It is mainly cultivated in subtropical regions where it rains a lot, and has various shapes such as oval and spindle, and can also be curved or branched. The main ingredients are curcumin, demethoxycurcumin, zedoarone, turmerone, and eugenol. In oriental medicine, it has been used for purposes such as removing blood clots 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 the East and West. For the production of turmeric extract in the present invention You can use the rhizome of Curcuma longa , purchase it commercially, or use it collected or cultivated from nature.

Maitake mushroom ( Grifola frondosa ) belongs to the king maitake and maitake genus. It is called maitake mushroom because it looks like ginkgo leaves are overlapped in several layers. It comes in black, white, and brown colors. Maitake mushrooms are known to contain particularly high amounts of beta-glucan, which is commonly found in mushrooms, and were approved as an anti-cancer supplement by the U.S. FDA in 1998, and have been clinically reported to have anti-cancer effects. . It is also known as a natural plant with anti-cancer effects, suppressing blood sugar levels, lowering cholesterol, and suppressing aging. In order to produce maitake mushroom extract in the present invention, the fruiting body of Grifola frondosa can be used, commercially sold ones can be purchased and used, or those collected or cultivated in nature can be used.

Cornus officinalis ( Cornus) officinalis ) belongs to the Dogwood genus of the Dogwood family, and its fruits are mainly used. It has a long oval shape and is green at first, then changes to dark red-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, which strengthen the liver and kidneys, protect the waist and knees, and improve sexual function. It is a natural plant with efficacy. In order to produce Cornus officinalis extract in the present invention, the fruits of Cornus officinalis may be used, those sold commercially may be purchased and used, or those collected or cultivated in nature may be used.

The active ingredient included in the composition for enhancing immunity of the present invention can have a significant synergistic effect in terms of enhancing immunity compared to each single substance by combining two or more types of herbal extracts. The combination of two or more of the above herbal extracts is preferably (i) Piper longum extract or Poncirus trifoliata extract; and (ii) red ginseng extract or turmeric ( Curcuma longa ) extract.

In the present invention, the term "immunity enhancement" refers to all phenomena in which factors that increase immunity in the body are increased or factors that suppress immunity in the body are decreased by administration of a composition.

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

In the present invention, the term "extract" may be obtained by extraction with one or more solvents selected from the group consisting of water, C1 to C4 lower alcohol, 1,3-butylene glycol, and ethyl acetate, but is limited thereto. It doesn't work. Specifically, the manufacturing 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, maitake mushroom, or cornelian cherry can be used without limitation, whether cultivated or commercially available.

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, and is preferably may be fermented alcohol, but is not limited thereto. Extraction methods such as fermented alcohol extraction, immersion extraction, reflux cooling extraction, or ultrasonic extraction may be used, but are not limited to these. The extraction solvent can be extracted by adding 1 to 10 times the amount of dried Pilbal, Tangja, red ginseng, turmeric, maitake mushroom, or Cornus officinalis, 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. Additionally, the extraction time may be 1 hour to 48 hours, specifically 2 hours to 8 hours, but is not limited thereto. In addition, the number of extractions may be 1 to 5 times, and specifically, extraction may be repeated 1 to 3 times, but is not limited thereto.

The concentration step of the present invention may additionally include a process of concentrating the extract under reduced pressure, and the reduced pressure concentration may use 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 reduced pressure concentration process, and the drying may be reduced pressure 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.

A composition containing two or more extracts of the present invention described above as active ingredients can be used as a health functional food composition to improve immunity.

In the present invention, the term "food" includes, for example, various foods, beverages, gum, tea, vitamin complexes, health functional foods, etc., and includes all foods in the conventional sense. The food can be manufactured into dosage forms such as tablets, granules, powders, capsules, liquid solutions and pills according to known manufacturing methods, 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 from weight% to 100% by weight, but is not limited to this. Other than including the extract according to the present invention as an active ingredient, there are no particular restrictions on other ingredients, and various common flavoring agents or natural carbohydrates may be included as additional ingredients.

Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (e.g., 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 as food for special health use (FoSHU), which is a medicine processed to efficiently exhibit bioregulatory functions in addition to nutritional supply, and medical effects. It means high food. Here, “function” means adjusting nutrients to the structure and function of the human body or obtaining useful effects for health purposes, such as physiological effects. The food of the present invention can be manufactured by methods commonly used in the industry, and can be manufactured by adding raw materials and ingredients commonly added in the industry. Additionally, the food formulation can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms, and unlike general drugs, it is made from food as a raw material, so there are no side effects that may occur when taking the drug for a long time, and it has the advantage of being highly portable.

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 by encapsulating, powdering, or suspending it, and consuming it may cause certain health problems. It means bringing about an effect, but unlike regular drugs, it has the advantage of not having any side effects that may occur when taking the drug for a long time since it is made from food.

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 can be used.

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

In addition, the food composition contains preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxide) roxitoluene (BHT), etc.), colorants (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, saccharin) , sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), strengtheners, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents, improvers, etc. May contain food additives. The additives can be selected depending on the type of food and used in an appropriate amount.

As an example of the food composition of the present invention, it can be used as a health drink composition, and in this case, it can contain various flavoring agents or natural carbohydrates as additional ingredients, like ordinary drinks. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; polysaccharides such as dextrins and cyclodextrins; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as thaumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can 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 drink composition of the present invention.

In addition to the above, the health drink composition includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol or carbonating agent. Additionally, it may contain pulp for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients can be used independently or in combination. The ratio of the above components may be selected in the range of 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 are preferably contained in relevant laws and regulations of Korea, China, the United States, Europe, Japan, etc. (e.g., Food Code (Korea), Food Additives Code (Korea), and Health Functional Food Code ( Do not exceed the maximum usage value specified 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 permitted by the relevant laws and regulations of each country.

Additionally, a composition containing two or more extracts of the present invention as active ingredients can be used as a pharmaceutical composition to improve immunity.

In the present invention, the term “pharmaceutical composition” refers to a product prepared for the purpose of enhancing immunity, and can be formulated and used in various forms according to conventional methods.

The pharmaceutical composition of the present invention may contain one or more active ingredients that exhibit the same or similar functions in addition to the above ingredients. The pharmaceutical composition of the present invention may further include pharmaceutically acceptable additives, wherein the pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, and calcium hydrogen phosphate. , lactose, mannitol, taffy, 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, white sugar, dextrose, sorbitol, and talc 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 can be administered in various oral and parenteral dosage forms during actual clinical administration. When formulated, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants are used. It can be prepared using. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract of the present invention with at least one excipient, such as starch, calcium carbonate, and sucrose. It can be prepared by mixing sucrose, lactose, or gelatin. Additionally, in addition to simple excipients, lubricants such as magnesium styrate talc may also be used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Preparations for parenteral administration may include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used.

The pharmaceutical composition of the present invention can be administered orally or parenterally according to the desired method. When administered parenterally, it is administered externally through the skin or intraperitoneally, intrarectally, subcutaneously, intravenously, intramuscularly, or intrathoracically. You can choose the injection method. The dosage may vary depending on the patient's weight, age, gender, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

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

Additionally, a composition containing two or more extracts of the present invention as active ingredients can also be used as a quasi-drug composition to improve immunity.

In addition to the above ingredients, the quasi-drug composition of the present invention may further include pharmaceutically acceptable carriers, excipients, or diluents as needed. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effect of the present invention, and includes, for example, fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc. It can be included.

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., and the formulation method, dosage, usage method, and components of the quasi-drug are commonly known in the technical field. It can be appropriately selected from the techniques of.

It was confirmed that the mixture containing two or more herbal extracts of the present invention as active ingredients synergistically enhances interferon gamma when mixed and treated rather than when each extract is treated individually (Tables 1 to 8). Since interferon gamma plays an important role in immunity in the body, this composition can be used to improve immunity in the body.

Therefore, it was confirmed that a composition containing two or more herbal extracts as active ingredients 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.

The composition containing two or more herbal extracts of the present invention as active ingredients promotes the secretion of interferon gamma without being toxic to cells, so it can be usefully used to improve immunity.

Hereinafter, the present invention will be described in detail by the following examples and production 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 following examples and preparation examples. In addition, since these examples are only for the purpose of facilitating understanding of the present invention, the scope of the present invention is not limited by them in any way.

Example

I. Preparation of extract

Manufacturing example One: maitake mushroom , curcuma , Cornus officinalis, tangerine, red ginseng or It's a must Preparation of extract

Maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng or Pilbal extract was prepared by a manufacturing method including the following steps.

1) Washing and drying maitake mushrooms, turmeric, Cornus officinalis, tangerine, red ginseng, or pilbal, then pulverizing them into powder;

2) Adding fermented alcohol to 1000 kg of maitake mushroom, turmeric, Cornus officinalis, tangerine, 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 maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng, or Pilbal extract.

Manufacturing example 2: maitake mushroom , curcuma , Cornus officinalis, tangerine, red ginseng or It's a must Preparation of extract

Maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng or Pilbal extract was prepared by a manufacturing method including the following steps.

1) Washing and drying maitake mushrooms, turmeric, Cornus officinalis, tangerine, red ginseng, or pilbal, then pulverizing them into powder;

2) Adding water to 1000 kg of maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng or Pilbal powder obtained in step 1) and extracting at room temperature for 3 days;

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 maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng, or Pilbal extract.

Manufacturing example 3: maitake mushroom , curcuma , Cornus officinalis, tangerine, red ginseng or It's a must Preparation of extract

Maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng or Pilbal extract was prepared by a manufacturing method including the following steps.

1) Washing and drying maitake mushrooms, turmeric, Cornus officinalis, tangerine, red ginseng, or pilbal, then pulverizing them into powder;

2) Adding water to 1000 kg of maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng or Pilbal powder obtained in step 1) and extracting 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 maitake mushroom, turmeric, Cornus officinalis, tangerine, red ginseng, or Pilbal extract.

II. Evaluation of immunity-boosting effects

Example One: maitake mushroom extract and curcuma Efficacy of extract mixture to promote interferon gamma secretion

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

Interferon gamma (IFNγ) is one of the representative cytokines involved in body immunity, and is reported to play an important role in regulating immune responses to viruses, harmful bacteria, and fungi that cause disease 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 molds that cause disease in the human body is suppressed or killed, making it less likely to get sick.

The 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 various types of cytokines that cause natural killer cells to produce interferon gamma. There are macrophages that secrete. In the body, these two interact and secrete interferon gamma to regulate the body's immunity.

Accordingly, 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 maitake mushroom extract and turmeric extract at various concentrations, and then treated the macrophages with a mixture of maitake mushroom extract and turmeric extract. Macrophage culture medium containing cytokines secreted in response to natural killer cells is treated with natural killer cells, which are other cells mainly involved in the production and secretion of interferon gamma in the body, and then the concentration of interferon gamma secreted by natural killer cells is determined by enzyme binding. It was analyzed by immunoprecipitation assay (Enzyme-linked Immunosorbent Assay, ELISA) as follows.

The day before the experiment to analyze interferon gamma concentration, interferon gamma capture antibody was diluted to 2 μg/ml, then 100 ul per well was added to an immunoplate and left at room temperature. The next day, all capture antibodies were removed, washed three times with 1 During blocking, the culture medium of natural killer cells to analyze the concentration of interferon gamma was centrifuged and the supernatant was taken. 1 hour after starting blocking, all reagent diluent 2 was removed and washed three times with 1X wash buffer. 100 ul of the supernatant taken from above was added to each well and mixed at 300 rpm. At this time, in order to obtain the concentration-absorbance standard curve of interferon gamma, the interferon gamma standard solution was 9.375 ng/ml, 18.75 ng/ml, 37.5 ng/ml, 75 ng/ml, 150 ng/ml, 300 ng/ml, and 600 ng. It was diluted to /ml and added to the well next to the well containing the culture medium of natural killer cells, 100 ul per well. After 2 hours, all natural killer cell culture medium and interferon gamma standard solution were removed, washed three times with 1X wash buffer, and 100ul of detection antibody diluted to 0.125 μg/ml was added to each well and shaken at 300rpm. After 2 hours, all detection antibodies were removed, washed three times with 1 After 20 minutes, all streptoavidin-HRP was removed, 100 ul of substrate was added to each well, blocked from light, and shaken at 300 rpm for 5 minutes. After shaking for 5 minutes, it was confirmed that the color of the well containing the interferon gamma standard solution changed to blue in a concentration-dependent manner. 50ul of 2N sulfuric acid was added per well and the plate was shaken well to mix the contents evenly. Immediately after adding 2N sulfuric acid, the green color changed to yellow as it was mixed 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 the concentration of each interferon gamma. The negative control was converted to 1 in Table 1 below. It is described as one fold value.

Maitake 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 synergistically increased when the maitake mushroom extract and the turmeric extract were mixed to 100 μg/ml, compared to when the maitake mushroom extract and the turmeric extract were treated alone at 100 μg/ml. It has been done. In addition, when maitake mushroom extract and turmeric extract were mixed at a ratio of 50:50, the increase in interferon gamma concentration was the largest. As the ratio of turmeric extract increased, the increase in interferon gamma concentration gradually decreased. As the ratio of extract increased, the increase in interferon gamma concentration rapidly decreased (Table 1).

Therefore, it was found that maitake mushroom extract and turmeric extract synergistically increase interferon gamma concentration, resulting in a synergistic effect of improving immunity in the body.

Example 2: maitake mushroom Interferon gamma secretion-promoting effect of extract and Cornus officinalis extract mixture

The maitake mushroom extract was the same as that prepared in Example 1, and Cornus officinalis was extracted with water at 100°C for 2 hours using the fruit according to Preparation Example 3.

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 maitake mushroom extract and Cornus officinalis extract at various concentrations in the same manner as above, and then the macrophages treated the macrophages with the maitake mushroom extract and Cornus officinalis extract. The concentration of interferon gamma secreted from natural killer cells after treating macrophage culture medium containing cytokines secreted in response to the mixture of extracts with natural killer cells, which are other cells mainly involved in the production and secretion of interferon gamma in the body. was analyzed by enzyme-linked immunosorbent assay (ELISA).

The concentration of interferon gamma was calculated in the culture medium obtained by treating macrophages and natural killer cells with a mixture of maitake mushroom extract and Cornus officinalis extract in the same manner as above. The fold value was calculated by setting the negative control group to 1 in Table 2 below. It was described.

Maitake mushroom extract (ppm) 0 100 90 80 70 60 50 40 30 20 10 0 Cornus officinalis 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 interferon gamma concentration was synergistically greater when the Maitake mushroom extract and Cornus officinalis extract were mixed and treated to 100 μg/ml than when the Maitake mushroom extract and Cornus officinalis extract were treated alone at 100 μg/ml. In addition, the increase in interferon gamma concentration was the largest when the maitake mushroom extract and Cornus officinalis extract were mixed at a ratio of 50:50. When the ratio of Cornus officinalis extract or the ratio of Maitake mushroom extract increased, the increase in interferon gamma concentration was similar. decreased (Table 2).

Therefore, it was found that maitake mushroom extract and Cornus officinalis extract synergistically increase the concentration of interferon gamma, resulting in a synergistic effect of improving the body's immunity.

Example 3: maitake mushroom Interferon-gamma secretion-stimulating effect of extract and Tangja extract mixture

The maitake mushroom extract was the same as that prepared in Example 1, and the fruit of Tangja was extracted with 70% fermented alcohol at 60°C for 2 hours according to Preparation Example 1 above.

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 maitake mushroom extract and Tangja extract at various concentrations in the same manner as above, and then the macrophages reacted with the maitake mushroom extract and Tangja extract. The concentration of interferon gamma secreted from natural killer cells after treating macrophage culture medium containing cytokines secreted in response to the mixture of extracts with natural killer cells, which are other cells mainly involved in the production and secretion of interferon gamma in the body. was analyzed by enzyme-linked immunosorbent assay (ELISA).

Interferon gamma concentration was calculated in the culture medium obtained by treating macrophages and natural killer cells with a mixture of maitake mushroom extract and Tangerine extract in the same manner as above, and the fold value was calculated by setting the negative control group to 1 in Table 3 below. It was described.

Maitake 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 interferon gamma concentration was synergistically greater when the Maitake mushroom extract and Tangerine extract were mixed and treated to 100 μg/ml than when the Maitake mushroom extract and Tangerine extract were treated alone at 100 μg/ml. In addition, the increase in interferon gamma concentration was the largest when maitake mushroom extract and Tangerine extract were mixed in a ratio of 50:50. As the ratio of Tangia extract increased, the increase in interferon gamma concentration gradually decreased, and the increase in interferon gamma concentration decreased gradually. As the ratio of extract increased, the increase in interferon gamma concentration rapidly decreased (Table 3).

Therefore, it was found that maitake mushroom extract and Tangja extract synergistically increase interferon gamma concentration, resulting in a synergistic effect of improving body immunity.

Example 4: maitake mushroom Efficacy of extract and red ginseng extract mixture to promote interferon gamma secretion

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

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 maitake mushroom extract and red ginseng extract at various concentrations in the same manner as above, and then macrophages were treated with the maitake mushroom extract and red ginseng extract. The concentration of interferon gamma secreted from natural killer cells after treating macrophage culture medium containing cytokines secreted in response to the mixture of extracts with natural killer cells, which are other cells mainly involved in the production and secretion of interferon gamma in the body. was analyzed by enzyme-linked immunosorbent assay (ELISA).

The concentration of interferon gamma was calculated in the culture medium obtained by treating macrophages and natural killer cells with a mixture of maitake mushroom extract and red ginseng extract in the same manner as above, and the fold value calculated by setting the negative control group to 1 in Table 4 below. It was described.

Maitake 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 interferon gamma concentration was synergistically greater when the maitake mushroom extract and the red ginseng extract were mixed at 100 μg/ml than when the maitake mushroom extract and the red ginseng extract were treated alone at 100 μg/ml. In addition, when maitake mushroom extract and red ginseng extract were mixed at a ratio of 50:50, the increase in interferon gamma concentration was the largest. When the ratio of red ginseng extract or the ratio of maitake mushroom extract increased, the increase in interferon gamma concentration was similar. decreased (Table 4).

Therefore, it was found that maitake mushroom extract and red ginseng extract synergistically increase the concentration of interferon gamma, resulting in a synergistic effect of improving immunity in the body.

Example 5: Red ginseng extract and It's a must Efficacy of extract mixture to promote interferon gamma secretion

Red ginseng was extracted with 70% fermented alcohol at 60°C for 2 hours using rhizomes according to Preparation Example 1 above, and the pilbal fruit was crushed, extracted with DMSO, and then filtered.

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by treating macrophages and natural killer cells with a mixture of red ginseng extract and pilbal extract in the same manner as above and then culturing them. The fold converted by setting the negative control group to 1 is shown in Table 5 below. It is described 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, it was confirmed that adding Pilbal extract to the red ginseng extract increased the concentration of interferon gamma compared to the treatment of red ginseng alone, and thus the immunity-enhancing effect of red ginseng in the body was enhanced by Pilbal.

Example 6: Interferon gamma secretion promotion effect of red ginseng extract and Tangja extract mixture

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

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by treating macrophages and natural killer cells with a mixture of red ginseng extract and Tangerine extract in the same manner as above and then culturing them. The fold converted by setting the negative control group to 1 is shown in Table 6 below. It is described 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, it was confirmed that adding Tangerine root extract to red ginseng extract increased interferon gamma concentration compared to when red ginseng was treated alone, and thus the immune-enhancing effect of red ginseng in the body was enhanced by Tangerine root.

Example 7: curcuma extract and It's a must Efficacy of extract mixture to promote interferon gamma secretion

The turmeric extract was the same as that prepared in Example 1, and the fruit was crushed, extracted with DMSO, and then filtered.

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by treating macrophages and natural killer cells with a mixture of turmeric extract and pilbal extract in the same manner as above and then culturing them. The fold converted by setting the negative control group to 1 is shown in Table 7 below. It is described 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, it was found that the turmeric extract and the pilbal extract synergistically increase the concentration of interferon gamma, and thus have a synergistic effect in improving the body's immunity.

Example 8: curcuma Interferon-gamma secretion-promoting effect of the extract and Tangja extract mixture

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

The present inventors calculated the concentration of interferon gamma in the culture medium obtained by treating macrophages and natural killer cells with a mixture of turmeric extract and Tangerine extract in the same manner as above and then culturing them. The fold converted by setting the negative control group to 1 is shown in Table 8 below. It is described 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, it was confirmed that adding Tangja extract to turmeric extract increased the concentration of interferon gamma compared to treatment with turmeric alone, and thus the immune-enhancing effect of turmeric in the body was enhanced by Tangja.

III. Food sensory evaluation

A beverage was prepared as shown in Table 9 below, and 50 healthy men and women aged 18 to 60 years old were allowed to drink it and then sensory evaluation was performed.

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 evaluators installed individual booths with partitions so that they could concentrate without exchanging opinions, and a sink for mouthwash 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 using the following sensory evaluation scale, and the average value was calculated.

<Sensory evaluation scale>

Strong taste (5), Tasty (4), Average (3), Weak taste (2), Hard to taste (1)

The results of drinks 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 Required (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 Required (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 confirmed in Tables 10 and 11 above, the Pilbal extract alone had strong spicy, bitter, and sour tastes, and thus had low palatability as a food. In addition, it was confirmed that the red ginseng extract alone had a strong bitter and sour taste, and the turmeric extract alone had a strong spicy, bitter, and sour taste. However, when Pilbal extract was mixed with red ginseng or turmeric extract, loss of taste was observed in terms of pungency, bitterness, and sourness, and the taste as a food was greatly improved.

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

Red ginseng (ppm) 100 60 50 40 0 Tangerine (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 Tangerine (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 confirmed in Tables 12 and 13 above, the bitter and sour taste of the Tangja extract alone was particularly strong, and thus its palatability as a food was low. In addition, it was confirmed that red ginseng extract alone and turmeric extract alone had a strong bitter and sour taste. However, when Tangerine extract was mixed with red ginseng or turmeric extract, loss of taste in terms of bitterness and sourness was confirmed, and the taste as a food was greatly improved.

In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.

Claims (5)

(i) Piper longum extract or Poncirus trifoliata extract; and
(ii) red ginseng extract or turmeric (Curcuma longa) extract
A health functional food composition for improving immunity containing a combination of the following as an active ingredient,
The Pilbal extract is a DMSO (Dimethyl sulfoxide) extract of Pilbal fruit, the Tangja extract is an ethanol aqueous solution extract of Tangja fruit, the red ginseng extract is an ethanol aqueous solution extract of red ginseng rhizome, and the turmeric extract is an ethanol aqueous solution extract of Curcuma rhizome. ,
A health functional food composition wherein the active ingredient is any one selected from the group consisting of:
- Red ginseng extract and Tangerine extract are combined in a weight ratio of 40:60 to 10:90 (red ginseng extract: Tangerine extract),
- Turmeric extract and Tangerine extract combined at a weight ratio of 70:30 to 40:60 (turmeric extract: Tangerine extract), and
- A combination of turmeric extract and pilbal extract at a weight ratio of 80:20 to 10:90 (turmeric extract: pilbal extract). delete delete The health functional food composition according to claim 1, wherein the active ingredient promotes the secretion of interferon gamma. A food containing the health functional food composition according to any one of paragraphs 1 or 4.