Immunity-boosting composition containing two or more herbal extracts as active ingredients

Abstract

To provide a composition for enhancing immunity and a composition for promoting interferon-γ secretion, comprising two or more herbal extracts as active ingredients, where the combination of the two or more herbal extracts synergistically increases the secretion of interferon-γ promoting immunity in the body without being toxic to cells and thus can be usefully employed for enhancing immunity.SOLUTION: A composition for food with health claims for enhancing immunity comprises, as active ingredients, two or more selected from the group consisting of Grifola frondosa extract, Cornus officinalis extract, Poncirus trifoliata extract, Piper longum extract, Curcuma longa extract, and Panax ginseng extract.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a composition for enhancing immunity containing two or more natural herbal extracts as active ingredients, and more specifically to a functional health food composition and pharmaceutical composition for enhancing immunity by increasing the secretion of interferon-gamma (IFNγ), which enhances immunity in the body. [Background technology]

[0002] Immunity is the ability of the human body to fight against external pathogens such as bacteria and viruses, killing or neutralizing them. Therefore, even when exposed to the same external pathogen, people with strong immunity will not become infected, while people with weak immunity will be susceptible to infection. Therefore, it can be said that strengthening immunity is of utmost importance in order to live a healthy life. Immunity can be divided into congenital immunity, which is born with us, and acquired immunity, which is acquired through exposure to external pathogens or vaccination.

[0003] Immunity first became known because those who survive an infectious disease are protected from the same disease. Immunity, first discovered around 430 BC, has since been extensively studied, leading to the development of various methods to enhance immunity. One of the most widely used methods is vaccination. Vaccination involves artificially injecting a weak infectious agent to stimulate the immune system, thereby strengthening immunity against that infectious agent. While vaccination is the most effective method currently developed to prevent infectious diseases, it has limitations, such as a narrow range of effects because it targets only one disease and the rare possibility of side effects. In fact, vaccinations are administered every year at the change of seasons to prevent influenza, which is caused by the influenza virus. However, because the prevalent influenza virus changes every year, vaccinations are ineffective unless the prevalent influenza virus for each year can be accurately predicted. Therefore, to overcome these limitations of vaccination, the body's own internal immunity must be enhanced to provide broad resistance to various infectious agents.

[0004] Cytokines are a collective term for proteins secreted by immune cells in the body and are important factors in regulating the body's immune system. Among these, interferon is known to primarily inhibit intracellular viral replication. Interferon was discovered in 1957 as a substance that inhibits viral growth. Interferon inhibits intracellular protein synthesis by producing protein kinase R (PKR), thereby preventing the synthesis of proteins necessary for the growth of viruses that have infected cells. It also inhibits viral growth by producing RNase L, which destroys viral RNA. Based on this principle, interferon is used to treat Middle East Respiratory Syndrome (MERS) and pandemic influenza. It is also known that bats, which carry many viruses, have high levels of interferon and therefore do not become ill. 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 suppress the growth of cancer cells. Interferon activates major histocompatibility complex (MHC) molecules I and II, activating cytotoxic T cells and helper T cells, which either directly recognize and kill infected cells or activate other immune cells.

[0005] Most immune-boosting drugs developed to date cite an increase in the number of immune cells as a mechanism for boosting immunity. However, even if the number of immune cells increases, if most of them are not activated, it cannot be said that immunity has increased. Therefore, it is also important to confirm the degree to which immune cells are activated. Interferon not only has the ability to boost the body's immune system itself, but can also be used as an indicator of various immune cell activities. Therefore, materials that can increase interferon levels in the body can be developed as materials for boosting the body's immune system.

[0006] In this specification, a number of documents are referenced and cited, and the disclosures of the cited documents are incorporated herein by reference in their entirety to more clearly describe the state of the art to which the present invention pertains and the content of the present invention. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Korean Patent No. 10-1967073 Summary of the Invention [Problem to be solved by the invention]

[0008] As a result of research efforts by the inventors to develop natural products that have immune-boosting effects without side effects, they discovered that combining two or more natural herbal extracts has a significant immune-boosting effect, leading to the completion of the present invention.

[0009] Therefore, an object of the present invention is to provide a functional health food composition for enhancing immunity, which contains two or more herbal extracts as active ingredients.

[0010] Another object of the present invention is to provide a pharmaceutical composition for enhancing immunity, which contains two or more herbal extracts as active ingredients.

[0011] Other objects and advantages of the present invention will be more clearly understood from the following detailed description of the invention, the claims and the drawings. [Means for solving the problem]

[0012] One aspect of the present invention provides a composition for enhancing immunity, comprising two or more active ingredients selected from the group consisting of Maitake mushroom (Grifola frondosa) extract, Cornus officinalis extract, Trifoliate orange (Poncirus trifoliata) extract, Long pepper (Piper longum) extract, Turmeric (Curcuma longa) extract, and Red ginseng (Panax ginseng) extract.

[0013] In one embodiment of the present invention, the active ingredients include a combination of (i) Maitake mushroom extract and (ii) turmeric extract, Cornus fruit extract, or Trifoliate orange extract, and may further include red ginseng extract.

[0014] In another embodiment of the present invention, the active ingredient may include a combination of (i) a long pepper extract or a trifoliate orange extract and (ii) a red ginseng extract or a turmeric extract.

[0015] The present invention will be specifically described below.

[0016] Maitake mushrooms (Grifola frondosa) belong to the genus Grifola in the family Grifolaraceae. They are called maitake because of their appearance of overlapping ginkgo-like structures. They come in black, white, and brown. They are known to contain particularly high amounts of β-glucan, a compound commonly found in mushrooms. They were approved by the U.S. FDA in 1998 as an anticancer adjuvant and have been clinically reported to have anticancer efficacy. In addition to their anticancer efficacy, they are also known as a natural plant with blood sugar suppression, cholesterol reduction, and anti-aging effects. In the present invention, maitake fruiting bodies may be used to produce maitake extract. Commercially available maitake mushrooms, as well as those harvested or cultivated from the wild, may be used.

[0017] Cornus officinalis (Cornus officinalis) belongs to the Cornaceae family and is primarily used for its fruit. It is oblong and initially green, turning a deep reddish-purple or purple as it ripens, becoming glossy and coarsely wrinkled. Its main components are ursolic acid, oleanolic acid, loganin, quercetin, kaempferol, and organic acids. It is a natural plant with the effects of strengthening the liver and kidneys, protecting the lower back and knees, and improving sexual function. In the present invention, Cornus officinalis fruit may be used to prepare Cornus officinalis extract. Commercially available Cornus officinalis fruit may be used, or Cornus officinalis fruit may be harvested or cultivated.

[0018] Trifoliate orange (Poncirus trifoliata) is a deciduous tree belonging to the genus Trifoliate orange in the family Rutaceae. It is spherical, with a dark brown or brown exterior and a light gray-brown interior. The immature fruit is primarily used as a medicinal ingredient. In traditional Chinese medicine, it is primarily used to clear phlegm and eliminate congestion, and is also used as a painkiller, antipyretic, and diuretic. Its main components are flavonoid glycosides, vitamin C (ascorbic acid), and acetic acid. In the present invention, immature trifoliate orange fruits may be used to produce a trifoliate orange extract. Commercially available fruits, naturally harvested fruits, or cultivated fruits may be used.

[0019] Piper longum is a plant belonging to the Piperaceae family, genus Piper. It has a similar smell to pepper but a more bitter taste. It is cylindrical, with numerous small granular fruits attached around the stalk. It is reddish-brown or black-brown in color. In traditional Chinese medicine, it is used to treat headaches, vomiting, diarrhea, etc. Its main components include piperlongumine, pipercide, piperine, sylvatine, and piplartine. In the present invention, dried long pepper fruits may be used to produce long pepper extract. Commercially available fruits, as well as fruits harvested or cultivated from the wild, may be used.

[0020] Turmeric (Curcuma longa) is a perennial herb of the Zingiberaceae family, primarily grown from rhizomes. It is primarily cultivated in subtropical regions with high rainfall and has a variety of shapes, including oval and spindle-shaped, and some are curved or branched. Its main components include curcumin, demethoxycurcumin, zedoarone, turmerone, and eugenol. In traditional Chinese medicine, it is used to remove blood stasis and improve digestive function. Turmeric has long been used as a medicinal ingredient and spice in both Eastern and Western cultures. In the present invention, turmeric rhizomes may be used to produce turmeric extract. These rhizomes may be commercially available, or may be harvested or cultivated in the wild.

[0021] Red ginseng (Panax ginseng) is the steamed root of Panax ginseng, a perennial herb belonging to the Araliaceae family. It has a long, slender, cylindrical-to-spindle shape, and some have numerous lateral roots emerging from the middle. The exterior is light gray-brown, while the interior is lighter brown. Its main component is ginsenoside, and while the type varies depending on the position and number of sugars attached, red ginseng is characterized by its high content of ginsenoside Rg3. It is used for purposes such as hangover relief, anti-aging, fatigue recovery, stress relief, immune enhancement, and male infertility treatment. In the present invention, commercially available red ginseng, as well as wild-harvested or cultivated red ginseng, may be used to prepare red ginseng extract.

[0022] The active ingredients contained in the immunity-boosting composition of the present invention can exert a significant synergistic effect in terms of immunity enhancement by combining two or more of the above-mentioned herbal extracts, compared to each substance alone.

[0023] In a preferred embodiment, the combination of two or more herbal extracts may be a combination of (i) Maitake mushroom extract and (ii) turmeric extract, Cornus fruit extract, or Trifoliate orange extract.

[0024] In another preferred embodiment, the combination of two or more herbal extracts may be a combination of (i) a long pepper extract or a trifoliate orange extract and (ii) a red ginseng extract or a turmeric extract.

[0025] In the present invention, the term "immunity enhancement" refers to any phenomenon in which factors that enhance immunity in the body are increased or factors that suppress immunity in the body are decreased by administration of a composition.

[0026] The extracts of natural herbal ingredients such as Maitake mushroom, Cornus japonica, Trifoliate orange, Long pepper, Turmeric, and Red ginseng may include simple water extracts, organic solvent extracts, solvent fractions of the extracts, and polysaccharide fractions, or may be mixtures or composites of the simple water extracts, organic solvent extracts, solvent fractions, and polysaccharide fractions.

[0027] In the present invention, the term "extract" may refer to a substance obtained by extraction with one or more solvents selected from the group consisting of water, C1-C4 lower alcohols, 1,3-butylene glycol, and ethyl acetate, but is not limited thereto. Specifically, the process for producing the extract of the present invention broadly includes an extraction step, a concentration step, and a drying step. In the process for producing the extract, Maitake mushroom, Cornus cherry, Trifoliate orange, Pepper, Turmeric, and Red ginseng can be used without limitation, whether they are cultivated or commercially available.

[0028] In the extraction step of the present invention, the extraction solvent may be one or more solvents selected from the group consisting of water, C1-C4 lower alcohols, 1,3-butylene glycol, and ethyl acetate, preferably fermented alcohol, but is not limited thereto. The extraction method may be, but is not limited to, fermented alcohol extraction, immersion extraction, reflux cooling extraction, or ultrasonic extraction. The extraction solvent may be added to dried Maitake mushroom, Cornus japonica, Trifoliate orange, Long pepper, Turmeric, or Red ginseng in a 1:1 to 10:1 ratio, and the extraction temperature may be 20°C to 100°C, specifically, but is not limited to, 50°C to 80°C. The extraction time may be 1 hour to 48 hours, specifically, but is not limited to, 2 hours to 8 hours. The number of extractions may be 1 to 5 times, specifically, 1 to 3 times, but is not limited thereto.

[0029] The concentrating step of the present invention may further include a process of concentrating the extract under reduced pressure, and the vacuum concentration may be performed using a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto.

[0030] The drying step of the present invention further includes a step of drying the extract obtained by the vacuum concentration process, and the drying may be, but is not limited to, vacuum drying, vacuum drying, boiling drying, spray drying, or freeze drying.

[0031] The extract of the present invention may enhance the human immune system by increasing the secretion of interferon-γ through the activation of immune cells.

[0032] A composition containing two or more of the above-mentioned extracts of the present invention as active ingredients can be used as a functional health food composition for enhancing immunity.

[0033] In the present invention, the term "food" refers to, for example, various foods, beverages, gums, teas, vitamin complexes, and health functional foods, and includes all foods in the usual sense. The food can be prepared in dosage forms such as tablets, granules, powders, capsules, liquid solutions, and pills using known preparation methods, and the content of the extract according to the present invention can be adjusted to, but is not limited to, 0.0001% by weight to 100% by weight based on the total weight of the food composition depending on the dosage form. In addition to the extract according to the present invention as an active ingredient, there are no particular limitations on other ingredients, and various common flavorings or natural carbohydrates may be included as additional ingredients.

[0034] Examples of the natural carbohydrates include common sugars such as monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), polysaccharides (e.g., dextrin, cyclodextrin, etc.), and sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). Other flavoring agents that can be preferably used include natural flavoring agents (thaumatin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (saccharin, aspartame, etc.). The proportion of the natural carbohydrates per 100 ml of the composition of the present invention can generally be about 1 g to 20 g, specifically about 5 g to 12 g.

[0035] In the present invention, the term "health functional food" is the same as "food for special health use (FoSHU)" and refers to a food with high medical and therapeutic effects that is processed to efficiently perform bioregulatory functions in addition to providing nutrients. Here, "functional" refers to obtaining useful health effects, such as regulating nutrients or physiological effects on the structure and function of the human body. The food of the present invention can be prepared by methods commonly used in the art and can be prepared by adding raw materials and ingredients commonly added in the art. Furthermore, the dosage form of the food is not particularly limited as long as it is a dosage form that is recognized as a food. The food composition of the present invention can be prepared in various dosage forms, and unlike general medicines, it is made from food ingredients, so it is free from side effects that can occur with long-term drug administration and has the advantages of being highly portable.

[0036] Specifically, the functional health food refers to a food product prepared by adding the composition of the present invention to food ingredients such as beverages, teas, spices, gum, and confectioneries, or by encapsulating, powdering, or suspending the composition, which provides specific health benefits when ingested. However, unlike general medicines, the functional health food has the advantage of being free from side effects that can occur when taking medicines for a long period of time, since it is made from food ingredients.

[0037] The food composition may further contain a physiologically acceptable carrier, but the type of carrier is not particularly limited, and any carrier commonly used in the art may be used.

[0038] The food composition may also contain additional ingredients commonly used in food compositions to improve smell, taste, visual appearance, etc. Examples include vitamin A, vitamin C, vitamin D, vitamin E, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, biotin, folate, panthotenic acid, etc. Minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu) may also be included; and amino acids such as lysine, tryptophan, cysteine, and valine.

[0039] The food compositions may also contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high-strength bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color formers (sodium nitrite, sodium acetate, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, sodium saccharin, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium D-bitartrate, etc.), strengtheners, emulsifiers, thickeners (thickeners), coating agents, gum bases, foam inhibitors, solvents, and improvers. The additives may be selected and used in appropriate amounts depending on the type of food.

[0040] An example of a food composition according to the present invention is a health drink composition, which, like conventional beverages, may contain various flavorings or natural carbohydrates as additional ingredients. The natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; or sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of sweeteners that can be used include natural sweeteners such as thaumatin and stevia extract; and synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates per 100 ml of the health drink composition of the present invention may generally be about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g.

[0041] The health drink composition may also contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonation agents. Fruit pulp may also be included for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in combination. The ratio of the above ingredients may be selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

[0042] Preferably, all ingredients described in the present invention do not exceed the maximum use values ​​stipulated in relevant laws, regulations, and standards of Korea, China, the United States, Europe, Japan, etc. (e.g., Food Codex (Korea), Food Additives Codex (Korea), Health Functional Food Codex (Korea)). That is, preferably, the food or food composition according to the present invention contains the ingredients according to the present invention in amounts within the limits permitted by the relevant laws, regulations, and standards of each country.

[0043] Furthermore, a composition containing two or more extracts of the present invention as active ingredients can also be used as a pharmaceutical composition for enhancing immunity.

[0044] In the present invention, the term "pharmaceutical composition" means a substance prepared for the purpose of enhancing immunity, and can be formulated into various forms by conventional methods for use.

[0045] The pharmaceutical composition of the present invention may further contain one or more active ingredients that perform the same or similar functions as the above-mentioned ingredients. The pharmaceutical composition of the present invention may further contain pharmaceutically acceptable additives, including starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, candy, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry®, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, and talc. The pharmaceutically acceptable additives of the present invention may be present in an amount of 0.1 to 90 parts by weight of the composition, but are not limited thereto.

[0046] The pharmaceutical composition of the present invention can be administered in various oral and parenteral dosage forms. Conventional diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, can be used in formulations. Solid formulations for oral administration include tablets, pills, powders, granules, and capsules. These solid formulations can be prepared by mixing the extract of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin. Lubricants, such as magnesium stearate and talc, can also be used in addition to simple excipients. Oral liquid formulations include suspensions, oral solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, can be used. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspension solvents that can be used include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. Suppository bases that can be used include Witepsol (registered trademark), Macrogol, Tween 61, cacao butter, laurin butter, glycerol gelatin, etc.

[0047] The pharmaceutical composition of the present invention can be administered orally or parenterally depending on the intended method, and parenteral administration can be performed by topical application to the skin or by injection such as intraperitoneal, rectal, subcutaneous, intravenous, intramuscular, or intrathoracic injection. The dosage range may vary depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease.

[0048] The dosage of the pharmaceutical composition of the present invention varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease. The daily dosage may be 0.0001 mg / kg to 100 mg / kg, specifically 0.001 mg / kg to 10 mg / kg, based on the amount of the extract of the present invention, and may be administered once to six times a day, but is not limited thereto.

[0049] Furthermore, a composition containing two or more extracts of the present invention as active ingredients can also be used as a quasi-drug composition for enhancing immunity.

[0050] In addition to the above ingredients, the quasi-drug composition of the present invention may further contain a pharmaceutically acceptable carrier, excipient, or diluent, as needed. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effects of the present invention, and may include, for example, a filler, extender, binder, wetting agent, disintegrant, surfactant, lubricant, sweetener, flavoring agent, preservative, etc.

[0051] The quasi-drug composition of the present invention includes, but is not limited to, disinfectants, cleansers, shower gels, ointments, wet tissues, coating agents, etc., and the formulation method, dosage, application method, components, etc. of the quasi-drug can be appropriately selected from conventional techniques known in the art.

[0052] It was confirmed that a mixture containing two or more herbal extracts of the present invention as active ingredients synergistically enhanced interferon-γ when mixed and treated compared to when each extract was treated alone (Tables 1 to 8). Since interferon-γ plays an important role in the body's immune system, the composition can be used to enhance the body's immune system.

[0053] Therefore, it has been confirmed that a composition containing two or more types of 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. [Effects of the Invention]

[0054] The composition containing two or more herbal extracts of the present invention as active ingredients is non-toxic to cells and promotes the secretion of interferon-γ, so it can be useful for enhancing immunity. DETAILED DESCRIPTION OF THE INVENTION

[0055] The present invention will be described in detail below with reference to examples and preparation examples. However, the following examples and preparation examples are merely illustrative of the present invention and are not intended to limit the scope of the present invention. Furthermore, the following examples are provided merely to aid in understanding the present invention and are not intended to limit the scope of the present invention in any way. [Example]

[0056] I. Extract Preparation Production Example 1: Production of Maitake mushroom, turmeric, Cornus japonica, Trifoliate orange, red ginseng or long pepper extract Extracts of Maitake mushroom, turmeric, Cornus cherry, Trifoliate orange, red ginseng or long pepper were prepared by a method including the following steps. 1) washing and drying Maitake mushroom, turmeric, Cornus japonica, Trifoliate orange, red ginseng or long pepper, and then grinding them into powder; 2) adding fermented alcohol to 1000 kg of the maitake mushroom, turmeric, Chinese quince, trifoliate orange, red ginseng or long pepper 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 cherry, Trifoliate orange, red ginseng or long pepper extract.

[0057] Production Example 2: Production of Maitake mushroom, turmeric, Cornus cherry, Trifoliate orange, red ginseng or long pepper extract Extracts of Maitake mushroom, turmeric, Cornus cherry, Trifoliate orange, red ginseng or long pepper were prepared by a method including the following steps. 1) washing and drying Maitake mushroom, turmeric, Cornus japonica, Trifoliate orange, red ginseng or long pepper, and then grinding them into powder; 2) adding water to 1000 kg of the maitake mushroom, turmeric, Chinese quince, trifoliate orange, red ginseng or long pepper powder obtained in step 1), and extracting the powder 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 cherry, Trifoliate orange, red ginseng or long pepper extract.

[0058] Production Example 3: Production of Maitake mushroom, turmeric, Cornus cherry, Trifoliate orange, red ginseng or long pepper extract Extracts of Maitake mushroom, turmeric, Cornus cherry, Trifoliate orange, red ginseng or long pepper were prepared by a method including the following steps. 1) washing and drying Maitake mushroom, turmeric, Cornus japonica, Trifoliate orange, red ginseng or long pepper, and then grinding them into powder; 2) adding water to 1000 kg of the maitake mushroom, turmeric, Chinese quince, trifoliate orange, red ginseng or long pepper 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 cherry, Trifoliate orange, red ginseng or long pepper extract.

[0059] II. Evaluation of immune-enhancing effects Example 1: Interferon-γ secretion promoting effect of a mixture of Maitake mushroom extract and turmeric extract Maitake mushroom mycelium was extracted for two days in water at room temperature according to Production Example 2, and turmeric rhizomes were extracted for two hours in 70% fermented alcohol at 60°C according to Production Example 1.

[0060] Interferon-γ (IFNγ) is one of the major cytokines involved in the body's immune system and has been reported to play an important role in regulating the immune response to viruses, harmful bacteria, fungi, etc. that cause disease in the human body. Therefore, if the secretion of interferon-γ increases, the body's immunity will increase, and the growth of viruses, harmful bacteria, fungi, etc. that cause disease in the human body will be suppressed or killed, resulting in less illness.

[0061] The cells that are primarily responsible for the production and secretion of interferon-γ in the body are natural killer cells (NK cells), which produce interferon-γ, and macrophages, which secrete various cytokines so that natural killer cells can produce interferon-γ. These two cells interact with each other in the body to secrete interferon-γ and regulate the body's immune system.

[0062] Therefore, the inventors treated macrophages, which are one of the cells mainly involved in the production and secretion of interferon-γ in the body, with a mixture of Maitake mushroom extract and turmeric extract at various concentrations, and then treated the macrophage culture medium containing cytokines secreted by the macrophages in response to the mixture of Maitake mushroom extract and turmeric extract with apoptotic cells, which are other cells mainly involved in the production and secretion of interferon-γ in the body, and analyzed the concentration of interferon-γ secreted by apoptotic cells using an enzyme-linked immunosorbent assay (ELISA) as follows.

[0063] The day before the interferon-γ concentration assay, the interferon-γ capture antibody was diluted to 2 μg / ml and added at 100 μl per well to the immunoplate and left at room temperature. The next day, the capture antibody was removed, and the plate was washed three times with 1X wash buffer. 300 μl of 1X diluted reagent diluent 2 was added per well to block any areas not covered by the capture antibody. During blocking, the culture medium of the naturally occurring cells to be assayed for interferon-γ concentration was centrifuged and the supernatant was collected. One hour after the start of blocking, the reagent diluent 2 was removed, and the plate was washed three times with 1X wash buffer. 100 μl of the collected supernatant was added per well and mixed at 300 rpm. To obtain an interferon-γ concentration-absorbance standard curve, interferon-γ standard solution was diluted to 9.375 ng / ml, 18.75 ng / ml, 37.5 ng / ml, 75 ng / ml, 150 ng / ml, 300 ng / ml, and 600 ng / ml and added at 100 μl per well to wells adjacent to the wells containing the natural killer cell culture medium. After 2 hours, the natural killer cell culture medium and interferon-γ standard solution were removed, and the wells were washed three times with 1X wash buffer. Then, 100 μl of detection antibody diluted to 0.125 μg / ml was added per well and shaken at 300 rpm. After 2 hours, the detection antibody was removed, the wells were washed three times with 1X wash buffer, and 100 μl of streptavidin-HRP diluted to 1X was added per well and shaken at 300 rpm in the dark. After 20 minutes, all streptavidin-HRP was removed, and 100 μl of substrate was added per well. The plate was then protected from light and shaken at 300 rpm for 5 minutes. After 5 minutes of shaking, the wells containing the interferon-γ standard solution were confirmed to have turned blue in a concentration-dependent manner. 50 μl of 2N sulfuric acid was then added per well, and the plate was vigorously shaken to mix the contents uniformly. The color was green immediately after adding the 2N sulfuric acid, but turned yellow as it mixed.The absorbance was then measured at 450 nm and 570 nm, and the absorbance of each well was substituted into the interferon-γ concentration-absorbance standard curve obtained from the standard solution to calculate each interferon-γ concentration. The concentrations were then listed in Table 1 as fold values ​​converted from the negative control group to 1.

[0064] [Table 1]

[0065] As a result, it was confirmed that the increase in interferon-γ concentration was synergistically greater when maitake extract and turmeric extract were mixed at 100μg / ml than when either maitake extract or turmeric extract was used alone at 100μg / ml. Furthermore, the increase in interferon-γ concentration was greatest when maitake extract and turmeric extract were mixed at a 50:50 ratio, and the increase in interferon-γ concentration gradually decreased as the ratio of turmeric extract increased, and the increase in interferon-γ concentration rapidly decreased as the ratio of maitake extract increased (Table 1).

[0066] Therefore, it was found that the Maitake mushroom extract and the turmeric extract synergistically increase the concentration of interferon-γ, and therefore have a synergistic effect of enhancing the body's immune system.

[0067] Example 2: Interferon-γ secretion promoting effect of a mixture of Maitake mushroom extract and Cornus japonica extract The same Maitake mushroom extract as prepared in Example 1 was used, and the fruit of the Cornus japonica was extracted with water at 100°C for 2 hours according to Preparation Example 3.

[0068] According to the above-mentioned method, the inventors treated macrophages, which are one of the cells mainly involved in the production and secretion of interferon-γ in the body, with a mixture of Maitake mushroom extract and Cornus cherry extract at various concentrations. Then, the macrophage culture medium containing cytokines secreted by the macrophages in response to the mixture of Maitake mushroom extract and Cornus cherry extract was treated with apoptotic cells, which are other cells mainly involved in the production and secretion of interferon-γ in the body, and the concentration of interferon-γ secreted by the apoptotic cells was analyzed by enzyme-linked immunosorbent assay (ELISA).

[0069] The interferon-γ concentration in the culture medium obtained by treating macrophages and natural killer cells with a mixture of Maitake mushroom extract and Cornus japonica extract as described above and culturing them was calculated and shown in Table 2 as a multiplier value converted from the negative control group to 1.

[0070] [Table 2]

[0071] The results showed that the increase in interferon-γ levels was synergistically greater when the maitake and Cornus cherry extracts were mixed at 100μg / ml than when either maitake or Cornus cherry extract was used alone at 100μg / ml. Furthermore, the increase in interferon-γ levels was greatest when the maitake and Cornus cherry extracts were mixed at a 50:50 ratio, and the increase in interferon-γ levels decreased similarly as the ratio of Cornus cherry extract or maitake extract increased (Table 2).

[0072] Therefore, it was found that the Maitake mushroom extract and the Cornus fruit extract synergistically increase the interferon-γ concentration, and therefore have a synergistic effect of enhancing the body's immune system.

[0073] Example 3: Interferon-γ secretion promoting effect of a mixture of Maitake mushroom extract and Trifoliate orange extract The same Maitake mushroom extract as prepared in Example 1 was used, and trifoliate orange fruit was extracted with 70% fermented alcohol at 60°C for 2 hours according to Production Example 1.

[0074] According to the above-mentioned method, the present inventors treated macrophages, which are one of the cells mainly involved in the production and secretion of interferon-γ in the body, with a mixture of Maitake mushroom extract and Trifoliate orange extract at various concentrations. Then, the macrophage culture medium containing cytokines secreted by the macrophages in response to the mixture of Maitake mushroom extract and Trifoliate orange extract was treated with spontaneously activated cells, which are other cells mainly involved in the production and secretion of interferon-γ in the body, and the concentration of interferon-γ secreted by the spontaneously activated cells was analyzed by enzyme-linked immunosorbent assay (ELISA).

[0075] The interferon-γ concentration in the culture medium obtained by treating macrophages and apoptotic cells with a mixture of Maitake extract and Trifoliate orange extract as described above and culturing them was calculated and shown in Table 3 as a multiplier value converted from the negative control group to 1.

[0076] [Table 3]

[0077] The results showed that the increase in interferon-γ levels was synergistically greater when the maitake and trifoliate orange extracts were mixed at 100μg / ml than when either maitake or trifoliate orange extract was used alone at 100μg / ml. Furthermore, the increase in interferon-γ levels was greatest when the maitake and trifoliate orange extracts were mixed at a 50:50 ratio. The increase in interferon-γ levels gradually decreased as the ratio of trifoliate orange extract increased, and the increase in interferon-γ levels rapidly decreased as the ratio of maitake extract increased (Table 3).

[0078] Therefore, it was found that the Maitake mushroom extract and the Trifoliate orange extract synergistically increase the interferon-γ concentration, and therefore have a synergistic effect of enhancing the body's immune system.

[0079] Example 4: Interferon-γ secretion promoting effect of a mixture of Maitake mushroom extract and red ginseng extract The same Maitake mushroom extract as prepared in Example 1 was used, and red ginseng rhizomes were extracted with 70% fermented alcohol at 60°C for 2 hours according to Preparation Example 1.

[0080] According to the above-mentioned method, the inventors treated macrophages, which are one of the cells mainly involved in the production and secretion of interferon-γ in the body, with a mixture of Maitake mushroom extract and red ginseng extract at various concentrations. Then, the macrophage culture medium containing cytokines secreted by the macrophages in response to the mixture of Maitake mushroom extract and red ginseng extract was treated with natural killer cells, which are other cells mainly involved in the production and secretion of interferon-γ in the body, and the concentration of interferon-γ secreted by the natural killer cells was analyzed by enzyme-linked immunosorbent assay (ELISA).

[0081] The interferon-γ concentration in the culture medium obtained by treating macrophages and apoptotic cells with a mixture of Maitake mushroom extract and red ginseng extract as described above and culturing them was calculated and shown in Table 4 as a multiplier value converted from the negative control group to 1.

[0082] [Table 4]

[0083] As a result, the increase in interferon-γ levels was synergistically greater when maitake extract and red ginseng extract were combined at 100μg / ml than when either maitake extract or red ginseng extract was used alone at 100μg / ml. Furthermore, the increase in interferon-γ levels was greatest when maitake extract and red ginseng extract were combined at a 50:50 ratio, and the increase in interferon-γ levels decreased similarly with increasing the ratio of red ginseng extract or maitake extract (Table 4).

[0084] Therefore, it was found that the Maitake mushroom extract and the red ginseng extract synergistically increase the concentration of interferon-γ, and therefore have a synergistic effect of enhancing the body's immune system.

[0085] Example 5: Interferon-γ secretion promoting effect of a mixture of red ginseng extract and long pepper extract Red ginseng roots were extracted with 70% fermented alcohol at 60°C for 2 hours according to Production Example 1, and long pepper fruit was crushed, extracted with dimethyl sulfoxide (DMSO), and then filtered.

[0086] The inventors calculated the interferon-γ concentration in the culture medium obtained by treating macrophages and apoptotic cells with a mixture of red ginseng extract and long pepper extract using the above-mentioned method, and the results are shown in Table 5 as a multiplier value converted from the negative control group to 1.

[0087] [Table 5]

[0088] As a result of the experiment, it was confirmed that when red ginseng extract was combined with long pepper extract, the interferon-γ concentration increased compared to when red ginseng was used alone, and that the immune-boosting effect of red ginseng was enhanced by long pepper.

[0089] Example 6: Interferon-γ secretion promoting effect of a mixture of red ginseng extract and trifoliate orange extract Red ginseng rhizomes were extracted with 70% fermented spirit at 60°C for 2 hours according to Production Example 1, and trifoliate orange fruits were extracted with 70% fermented spirit at 60°C for 2 hours according to Production Example 1.

[0090] The present inventors treated macrophages and apoptotic cells with a mixture of red ginseng extract and trifoliate orange extract using the above-mentioned method, and then cultured the culture medium. The interferon-γ concentration in the culture medium was calculated and shown in Table 6 as a multiplier value calculated by setting the negative control group at 1.

[0091] [Table 6]

[0092] The results of the experiment showed that when trifoliate orange extract was added to red ginseng extract, the interferon-γ concentration increased compared to when red ginseng was used alone, confirming that the immune-boosting effect of red ginseng was enhanced by trifoliate orange.

[0093] Example 7: Interferon-γ secretion promoting effect of a mixture of turmeric extract and long pepper extract The turmeric extract used was the same as that prepared in Example 1, and the long pepper fruit was crushed, extracted with DMSO, and then filtered.

[0094] The inventors calculated the interferon-γ concentration in the culture medium obtained by treating macrophages and apoptotic cells with a mixture of turmeric extract and long pepper extract using the method described above, and the results are shown in Table 7 as a multiplier value converted from the negative control group to 1.

[0095] [Table 7]

[0096] As a result of the experiment, it was found that turmeric extract and long pepper extract synergistically increased the concentration of interferon-γ, and therefore had a synergistic effect of enhancing the body's immune system.

[0097] Example 8: Interferon-γ secretion promoting effect of a mixture of turmeric extract and trifoliate orange extract The turmeric extract used was the same as that prepared in Example 1, and trifoliate orange fruit was extracted with 70% fermented alcohol at 60°C for 2 hours according to Production Example 1.

[0098] The present inventors calculated the interferon-γ concentration in the culture medium obtained by treating macrophages and apoptotic cells with a mixture of turmeric extract and trifoliate orange extract using the above-mentioned method, and the results are shown in Table 8 as a multiplication factor converted from the negative control group to 1.

[0099] [Table 8]

[0100] The results of the experiment showed that when trifoliate orange extract was added to turmeric extract, the interferon-γ concentration increased compared to when turmeric was used alone, confirming that the immune system-boosting effect of turmeric was enhanced by trifoliate orange extract.

[0101] III. Sensory Evaluation of Food A beverage was prepared according to the composition shown in Table 9, and 50 healthy men and women aged 18 to 60 drank it, after which a sensory evaluation was performed.

[0102] [Table 9]

[0103] To allow the sensory evaluators to concentrate without exchanging opinions, individual booths with partitions were set up, each equipped with a sink for gargling. The temperature was maintained at 20-25°C, the humidity at 50-60%, and ventilation facilities were provided to remove odors. The results of the drinking were evaluated using the sensory evaluation scale below, and the average score was calculated.

[0104] <Sensory evaluation scale> Strong taste (5), Slightly strong taste (4), Average taste (3), Weak taste (2), Hard to detect taste (1)

[0105] The results of the beverages prepared by mixing the scaled long pepper extract with red ginseng or turmeric extract are shown in Tables 10 and 11, respectively.

[0106] [Table 10]

[0107] [Table 11]

[0108] As can be seen from Tables 10 and 11, the long pepper extract alone had strong pungent, bitter, and sour tastes, resulting in poor palatability as a food. Red ginseng extract alone also had strong bitter and sour tastes, while turmeric extract alone also had strong pungent, bitter, and sour tastes. However, when the long pepper extract was mixed with red ginseng or turmeric extract, the pungent, bitter, and sour tastes were actually lost, resulting in a significant improvement in the taste as a food.

[0109] Furthermore, the results of the beverages produced by mixing trifoliate orange extract with red ginseng or turmeric extract using the above scale are shown in Tables 12 and 13, respectively.

[0110] [Table 12]

[0111] [Table 13]

[0112] As can be seen from Tables 12 and 13, the trifoliate orange extract alone had a particularly strong bitter and sour taste, making it unpalatable as a food. It was also confirmed that the red ginseng extract and turmeric extract alone also had strong bitter and sour tastes. However, when the trifoliate orange extract was mixed with the red ginseng or turmeric extract, the bitter and sour tastes were actually lost, resulting in a significant improvement in the taste as a food.

[0113] The above examples are illustrative in all respects and should not be understood as limiting. The scope of the present invention should be interpreted as including all modifications and variations derived from the meaning and scope of the claims that follow, as well as equivalent concepts, from the detailed description.

Claims

1. A health functional food composition for enhancing immunity, comprising as active ingredients a combination of Maitake mushroom (Grifola frondosa) extract and Cornus officinalis extract, or a combination of Maitake mushroom (Grifola frondosa) extract and Trifoliate orange (Poncirus trifoliata) extract.

2. The health functional food composition for enhancing immunity according to claim 1, further comprising a red ginseng (Panax ginseng) extract.

3. A health functional food composition for enhancing immunity, comprising as active ingredients a combination of long pepper (Piper longum) extract and red ginseng (Panax ginseng) extract, a combination of long pepper (Piper longum) extract and turmeric (Curcuma longa) extract, a combination of trifoliate orange (Poncirus trifoliata) extract and red ginseng (Panax ginseng) extract, or a combination of trifoliate orange (Poncirus trifoliata) extract and turmeric (Curcuma longa) extract.

4. 4. The immune system enhancing health functional food composition according to claim 1, wherein the active ingredient promotes the secretion of interferon-γ.

5. An immunity-boosting food comprising the immunity-boosting health functional food composition according to any one of claims 1 to 4.

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