KR20200062771A - Composition for Enhancing Immunity - Google Patents

Abstract

The present invention relates to a composition for enhancing immunity which contains lupeol and daucosterol as active components for promoting the production of antibodies and increasing the activity of immune cells, has no side effects, and is safe for the human body. In addition, the composition of the present invention is safe for the human body and has little side effects even when being taken for a long time.

Description

Composition for enhancing immunity {Composition for Enhancing Immunity}

The present invention relates to a composition for promoting the production of antibodies and increasing the activity of immune cells by including lupeol and dacosterol as active ingredients, and having no side effects and safe for the human body.

Immunity is a self-defense system that exists in the body, and it is a process of recognizing and removing and metabolizing various substances or living things that the human body invades from the outside as a foreign substance to itself.

It defends itself from damage from external stimuli or from the invasion of pathogenic microorganisms, but can also damage its own tissues, such as inflammatory reactions. It is a function that restores to normal by controlling changes in immune function or reduces the range of changes, and is divided into suppression of immune function or enhancement of immune function. Mitigating hyperimmune reactions means suppressing an undesirably increased immune response, such as allergic reactions caused by adverse reactions to foreign substances or responses to modified autoantigens.

Factors affecting immune function include genetic and environmental factors, age, gender, psychological stress, nutritional status, dietary habits, and diseases. Health functional foods having the function of regulating the immune function are classified into the functions of enhancing the reduced immune function, the function of regulating the excessive immune function, and the function of regulating the autoimmune function to control the immune response to the self component. When the immune function decreases, hypersensitivity reactions may occur. These hypersensitivity reactions may include asthma, seasonal or same-year rhinitis, allergic sinusitis, conjunctivitis, allergies caused by food and drugs, atopic dermatitis, hives, and allergies due to bee sting. Appears.

In order to overcome the problem of impairment of immune function due to these various causes, various immune enhancers and therapeutic agents are used, but there are side effects problems, and those that are commercially available are used for treatment rather than prevention. It is possible, and there is a need for a composition for improving immunity suitable for prevention.

The present inventors selected lupeol and dacosterol as materials having excellent efficacy while screening a substance having an immune enhancing function based on this immune-related gene mechanism, and showed excellent immunity enhancing efficacy as lupeol. The present invention was completed by examining the synergistic effect of and docosterol.

This result was studied with the support of the High Value-added Food Technology Development Project of the Ministry of Food, Agriculture, Forestry and Fisheries Technology Planning and Evaluation (115036-3).

Korean Registered Patent No. 10-0838435 Korean Registered Patent No. 10-1859296 Korean Patent Publication No. 10-2015-0019565

The object of the present invention is to promote the production of antibodies, including lupeol (Lupeol) and docosterol (Daucosterol) as an active ingredient to promote immunity and increase the activity of immune cells to help improve immunity, and onset It can be taken at any time before, there is no side effect showing a preventive and curative effect to provide a composition for promoting immune safety to the human body.

In order to achieve the above object, the present invention provides a composition for enhancing immunity comprising Lupeol and Daucosterol as active ingredients.

In addition, it provides a functional food comprising the composition for enhancing immunity.

In addition, it provides a pharmaceutical composition comprising the composition for enhancing immunity.

The composition for enhancing immunity of the present invention includes a mixture of lupeol and dacosterol as an active ingredient to promote the production of antibodies and increase the activity of immune cells, thereby improving immunity. In particular, for the rapid recovery of the immune function of the patient with reduced immunity, the composition for enhancing immunity of the present invention can be used, and even for a semi-healthy person with reduced immunity or a normal person with no abnormality in immune function, the immunity is enhanced. In order to use the composition of the present invention. In addition, the composition of the present invention is safe for the human body and has little side effects even when taken for a long time.

Therefore, the composition according to the present invention can be easily used as a functional food that can be seen as an effect by continuous administration. In addition, since it can be obtained from natural plant extracts, it can be preferably used for subjects with concerns about drug side effects, such as children, the elderly, or pregnant women.

1 is a test result of NK1.1+/CD69+ immune cell activity in the spleen by ingestion of Lupeol and Dacosterol in an inflammatory bowel disease mouse model (IBD, Inflammatory bowel disease).
FIG. 2 shows the results of the test for confirming the activity of CD8+/CD69+ immune cells in the spleen by ingestion of Lupeol and Dacosterol in an inflammatory bowel disease mouse model (IBD, Inflammatory bowel disease).
FIG. 3 is a test result of confirming the activity of CD11c+/CD69+ immune cells in the mesenteric lymph node by ingestion of Lupeol and Dacosterol in an inflammatory bowel disease mouse model (IBD, Inflammatory bowel disease).
4 is a result of a test for confirming a change in the level of a decrease in the amount of free radicals that is increased upon treatment with lipopolysaccharide (LPS) in a mouse macrophage cell line (RAW264.7) by treatment with Lupeol and Dacosterol.

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

In one embodiment, the present invention provides a composition for enhancing immunity comprising a mixture of lupeol and daucosterol as an active ingredient.

The chemical structures of the rufeol and the docosterol are shown in Chemical Formulas 1 to 2 below.

<Formula 1>

<Formula 2>

The lupeol is found in various plants, such as mango, legumes, and dandelion roots, and can be extracted using extraction solvents and supercritical extracts commonly used in the art.

The luperol is a pharmaceutical effective triterpenoid-based material, and is known to exhibit antibacterial, antiviral, anti-inflammatory, and anti-cancer effects.

The daucosterol (Daucosterol) is found in a variety of plants, such as thorns gallop, ginseng, cheonma, burdock, astragalus, can be extracted using the commonly used organic solvent, water or mixtures thereof, preferably It can be extracted using an organic solvent.

The daucosterol is a kind of pharmacologically effective saponin, and is known to exhibit neuroprotective action, cancer cell suppression action, candidiasis relief, stem cell proliferation, and antibacterial action.

Examples of the extraction solvent used for the extraction of the rufeol and docosterol purified water; Lower alcohols having 1 to 10 carbon atoms such as methanol, ethanol, isopropyl alcohol, and butanol; Polyhydric alcohols having 2 to 30 carbon atoms such as glycerol, ethylene glycol, propylene glycol, and 1,3-butylene glycol; And hydrocarbon-based solvents having 1 to 30 carbon atoms such as petroleum ether, methyl acetate, ethyl acetate, benzene, hexane, methylene chloride, diethyl ether, dichloromethane, and chloroform are exemplified. In addition, one or two or more solvents may be used in combination.

In one embodiment, the mixture of the rufeol and the docosterol may be included in an amount of 0.0001 to 50% by weight, preferably 0.001 to 10.0% by weight, and less than 0.0001% by weight based on 100% by weight of the total composition based on solid content. The effect of enhancing immunity is negligible, and it is not preferable because the formulation is difficult to apply at a concentration of more than 50% by weight.

In one embodiment, the lupeol and the docosterol may be included in a weight ratio of 1:1 to 1:10, preferably a weight ratio of 1:4, and if it exceeds the weight ratio, the immune enhancing effect may be insignificant.

The composition according to the present invention relates to an enhancement of immune function, refers to a food contributing to enhancing the immune function of the deteriorated human body, and may also include a case of promoting normal immune function.

Biomarkers for confirming the functional enhancement of immune function include NK cells, cytokines (IFN-α,IFN-β,TNF-α,IL-1,IL-6,IL-12), phagocytosis, NO for innate immunity , iNOS, COX, complement system, etc., and for adaptive immunity, cytokines (IFN-γ,TNF-β,IL-2,IL-4,IL-5,IL-6,IL-9,IL-10,IL- 13), lymphocyte subpopulation ratio, and immunoglobulin. In addition, there are common cell survival rates (increased number of immune cells), splenocyte proliferation, and tissue weight (spleen, thymus) in both innate and adaptive immunity. The contents of each biomarker are as follows.

Cell survival rate means an increase in immune function as the number of immune cells increases.

NO (Nitric oxide) is produced by the action of inducible nitric oxide synthase (iNOS) enzymes in macrophages, and the generated NO inhibits the modification of predated microbial (pathogen) constituent molecules and the action of enzymes containing Fe-S. To show anti-microbial action. In addition, NO is secreted from activated macrophages or neutrophils, killing extracellular pathogens.

Inducible nitric oxide synthase (iNOS) oxidizes L-arginine to produce L-citrulline and NO, and iNOS is expressed by stimulation such as cytokines in almost all cells.

COX2 (Cyclooygenase-2) is an enzyme that converts arachidonic acid to prostaglandin E2 (PGE2), which is expressed by inflammation or immune stimulation, and is induced in response to various cytokine growth factors.

Phagocytosis, also called phagocytosis, is the action of macrophages, such as white blood cells, to phagocytosis to remove bacteria and foreign substances. Macrophages have receptors for various molecular substances, and these receptors interact with ligands and microorganisms. Provokes reactions, which enhance the gluttony of pathogens.

Cytokine (Cytokine) is a protein that mediates cell-to-cell signaling, including interluekin (IL), interferon (IFN), and tumor necrosis factors (TNF). It is mainly produced by activated macrophages or lymphocytes, and the types of cytokines secreted from TH1 auxiliary T lymphocytes are IL-1, IL-2, IL-12, IL-15, IL-18, IFN-γ, and TNF-α Etc., these are inflammatory cytokines that exhibit cytotoxicity and are involved in cell-mediated immune responses.

The lymphocyte subpopulation ratio is a white blood cell formed by the differentiation and maturation of hematopoietic stem cells into lymphoid hematopoietic stem cells, and is largely divided into B lymphocytes, T lymphocytes, and NK cells. It is morphologically difficult, including even observation. This distinction is made possible by using a cell surface marker (CD) antibody characteristic for each cell. In the case of these markers, lymphocytes are characteristically different, and NK cells are CD3, CD56, CD16, CD94, and CD161, and NK1.1 and B lymphocytes, which are receptors acting on the functions of NK cells, are CD10, CD19, CD20, CD21, and CD22. , CD23, CD24, CD40, etc., T lymphocytes express CD3, secondary T lymphocytes CD4, cytotoxic T lymphocytes CD8 as markers on the cell surface, and several hematopoietic stem cells, T lymphocytes, etc. Express. In addition, macrophages, monocytes, and dendritic cells express CD11b, CD11c, and the like as markers.

The spleen and thymus are tissues related to immune function, the spleen is the tissue involved in lymphocyte production, and the thymus is T lymphocyte differentiation and maturation. Therefore, the spleen thymic tissue weight is measured to see the systematic effect of immune-related tissue.

The spleen is a tissue that makes lymphocytes and destroys aging red blood cells. Therefore, it means that the immune function improves as the number of splenocytes isolated from the test animal ingesting the test substance increases.

Immunoglobulin (Ig) is a serum protein that plays a major role in humoral immunity, and is classified into IgA, IgD, IgE, IgG, and IgM.Immunoglobulin (IgM) is an immunoglobulin mainly measured by immune function enhancing functions. IgA accounts for about 20% of immunoglobulins in serum, and is present in mucus such as saliva and upper respiratory secretion, and is involved in the defense of mucosal infection. IgG is divided into four subgroups IgG1, IgG2, IgG3 and IgG4, and is involved in toxin or virus neutralization, sedimentation, and aggregation reactions. IgM is complement binding and has high cohesive reactivity. An increase in immunoglobulins (IgA, IgG, IgM) means enhancing immune function.

Complement is a protein contained in blood and lymph fluid. Complement has no immunological specificity and binds to a complex of antigen and antibody to hemolyze and lyse to remove bacteria and antigens that have passed through the physical barrier. Complement consists of more than 20 proteins, mainly measuring C3, C4, C5. Activation of complement is associated with various immune responses, such as activation of macrophages or lymphocytes and enhancement of immune function.

NK cells are white blood cells in the blood that are responsible for innate immunity, and recognize cells that are infected with viruses or stressed cells and kill them directly or secrete inflammatory cytokines and kill them. It can be judged whether or not the immune function is enhanced by changing the NK cell activity.

According to another aspect of the present invention, the present invention provides a functional food for enhancing immunity comprising the composition for enhancing immunity.

The food composition according to the present invention can be prepared in the form of a composition by mixing with a known active ingredient known to be effective in promoting immunity or maintaining immunity, functional food, nutritional supplement, health food ( health food and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art. For example, as healthy foods, it can be prepared in the form of tea, juice, and drinks containing Lupeol and Dacosterol as active ingredients, and Lupeol and Dacosterol. It can be taken by granulating, encapsulating or powdering the food composition as an active ingredient.

In the food composition of the present invention, mineral-containing compounds such as vitamins A, B, C, D, E, beta-carotene, Ca, Mg, and Zn are used as auxiliary ingredients together with the active ingredients of the lupeol and daucosterol. It can be used, and red ginseng, ginseng, mushroom mycelium, beta glucan, Angelica mixed extract, spirulina, etc., can be used by mixing materials known to help boost the immune system.

In addition, in addition to the above components, as a known additive, natural flavors such as plum, lemon, pineapple, or herb flavors, natural juices, fructose, which is a natural pigment such as chlorophyllin and flavonoids, and sweeteners , Honey, sugar alcohol, sugar, citric acid and sodium citrate.

In the food composition of the present invention, the preferred content of the active ingredients that are Lupeol and Dacosterol may include about 0.0001 to 50% by weight based on the total weight of the food. Preferably, when ingested in an amount of 0.01mg/kg/day to 10mg/kg/day, an effect of enhancing immunity, restoring fatigue, or maintaining immune homeostasis may be exhibited.

The present invention provides a functional food in a form selected from the group consisting of tablets, pills, granules, powders, capsules, sprays and liquids containing the food composition.

Functional food refers to food designed and processed to sufficiently exert bioregulatory functions on the living body, such as biological defense of food ingredients, control of biorhythms, and prevention and recovery of diseases.

The functional food preparation is not particularly limited except that the food composition of the present invention is included, and may contain additional ingredients required for each food as a conventional food-acceptable food supplement.

Tablets are powdery or crystalline substances that are pressed together and made into rounded discs or cone-shaped formulations that are easy to take and the dosage is accurate. The functional food tablet of the present invention is not limited thereto, but may include excipients such as lactose, starch, white sugar, mannitol, sorbitol, inorganic salts, magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, May contain a binder such as sodium carboxymethylcellulose and/or polyvinylpyrrolidine, optionally disintegrating or boiling mixtures such as starch, agar, alginic acid or its sodium salt and/or absorbents, colorants, flavoring agents, And sweeteners. The formulation can be prepared by conventional mixing, granulating or coating methods.

Granules are round, fine-grained formulations, and in the case of mixed preparations, they can always take a certain ratio and are easy to take. Colorants, fragrances, and the like may be added to the granules of the present invention, if necessary, and can be coated with a suitable coating agent. The granules of granules are preferably 20% of the premise having a size of 355um or more and 1400um or less. The liquid formulation refers to a liquid formulation, and a stabilizer, mating agent, and preservative may be added according to the needs of the present invention. In addition, the functional beverage composition of the present invention may further include a fruit extract for improving flavor. The fruit extract gives the advantage of using various active ingredients possessed by the fruit together with improving flavor. The fruit used in the fruit extract is, but is not limited to, pineapple, lemon, citrus, orange, apple, pear, grapefruit, apricot, strawberry, peach, melon, guava, lemon, plum, and at least one selected from the Can be. Other functional beverage compositions of the present invention may include known additives. As these known additives, vitamin amide, vitamin A, B, C, D, E, folic acid, etc. can be used as vitamins, and L-glycine, L-lysine, DL-methionine, and L- as amino acids. Valine, biotin, L-cysteine, L-cystine, L-tryptophan, L-threonine, L-phenylalanine, etc. can be used, and as herbal medicines, goji berry, agapi, baekchul, county green, red ginseng, red ginseng, ginseng, yin and yang, woowang , Hwangjeong, etc. can be used, mixed fruit flavor, apple flavor, yogurt flavor, drink flavor, etc. can be used. Sweeteners include baekbaek sugar, glucose, fructose, fructooligosaccharide, isomaltoligosaccharide, malto-oligosaccharide, Chitosan oligosaccharides, chitooligosaccharides, soybean oligosaccharides, xyloligosaccharides, isomerized saccharides (high fructose), invert sugar, aspartame, stevioside, sorbitol, mannitol, etc. can be used, and citric acid, DL-apple acid, and succinic acid are used as acidifying agents. As a stabilizer, polysorbate 20, polysorbate 40, polysorbate 80, etc. among sorbates may be used, and as a preservative, benzoic acid and derivatives thereof, dehydroacetic acid and salts thereof, sorbic acid, and Salts and the like can be used.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for enhancing immunity comprising the composition for enhancing immunity.

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically acceptable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant, excipient, disintegrant, sweetener, binder, coating agent, expander, lubricant, A lubricant or flavoring agent may be used.

The pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.

Formulations of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectables. For example, for formulation in the form of tablets or capsules, the active ingredient can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and colorants can also be included in the mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, trachocanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

Acceptable pharmaceutical carriers in compositions formulated as liquid solutions include, as sterile and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets.

Furthermore, it can be preferably formulated according to each disease or component using methods disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA by appropriate methods in the art.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, and preferably oral administration. .

Suitable dosages of the pharmaceutical compositions of the present invention vary by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity, Usually, a skilled physician can easily determine and prescribe a dose effective for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.01 to 10 mg/kg/day.

The pharmaceutical composition of the present invention is prepared in a unit dose form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by those skilled in the art to which the present invention pertains. Or it can be manufactured by incorporating into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of ex, powder, granule, tablet, or capsule, and may further include a dispersant or stabilizer.

Hereinafter, the present invention will be described in more detail through examples and comparative examples. However, the configuration shown in the drawings and examples described in this specification is only one of the most preferred embodiments of the present invention, and does not represent all of the technical spirit of the present invention, and can replace them at the time of this application. It should be understood that there may be equivalents and variations.

Example: Preparation of lufeol and daucosterol

The lupeol and daucosterol were purchased through Sigma-Aldrich and were subjected to the following experiment.

Experimental Example 1: Level change test of NK1.1+/CD69+ cells in the spleen by ingestion of Lupeol and Daucosterol in an inflammatory bowel disease mouse model (IBD, Inflammatory bowel disease)

Inflammatory bowel disease (IBD) is a mouse model in which inflammation of the intestine is caused by abnormalities in the immune system. The IBD mouse model is tested to the extent that inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, which are chronic inflammatory bowel diseases, are expressed when DSS (dextran sodium sulfate) is ingested, and these conditions are alleviated by ingestion of test substances. The immune-improving effect of a substance can be measured.

IBD mice were administered 10 mg/kg/day of Lupeol and Dacosterol, respectively, for each test group, and samples mixed with Lupeol and Dacosterol in a ratio of 1:4. The control group was orally administered daily for 14 days at 10 mg/kg/day. Dextran sodium sulfate (DSS) was administered orally on the 15th day to induce inflammatory bowel disease, and samples were administered to the mixed administration group of Lupeol, Daucosterol and Lupeol and Dowosterol 1:4 for 9 days, respectively. Was administered. After completion of the test, the immune organs of the IBD mouse were separated and the immune cell changes of each immune organ were measured.

In order to confirm the activity and number change in the cells of each immune organ isolated from the IBD mouse model, the antibodies against the cell surface molecules serving as the respective markers were reacted and analyzed through flow cytometry (FACS).

In the spleen, 10 mg/kg of the test group mixed with lupeol and dowsterol 1:4 compared to the test group of 10 mg/kg/day administered with Lupeol or Daucosterol alone. It was confirmed that in the test group administered per day, the NK cell marker NK1.1+/CD69+, which is an immune cell, increased at the highest level and increased to a level similar to the normal group (WT). As a result, synergism between rufeol and daucosterol was confirmed.

The results are shown in FIG. 1.

Experimental Example 2: Level change test of CD8+/CD69+ cells in the spleen by ingestion of Lupeol and Daucosterol in an inflammatory bowel disease mouse model (IBD, Inflammatory bowel disease)

In mice with inflammatory bowel disease (IBD), Lupeol and Daucosterol were administered at 10 mg/kg/day for each test group, and Lupeol and Daucosterol were 1:4. The mixed sample was orally administered to the control group at 10 mg/kg/day for 14 days daily. Dextran sodium sulfate (DSS) was administered orally on the 15th day to induce inflammatory bowel disease, and samples were administered to the mixed administration group of Lupeol, Daucosterol and Lupeol and Dowosterol 1:4 for 9 days, respectively. Was administered. After completion of the test, the immune organs of the IBD mouse were separated and the immune cell changes of each immune organ were measured.

In order to confirm the activity and number change in the cells of each immune organ isolated from the IBD mouse model, the antibodies against the cell surface molecules serving as the respective markers were reacted and analyzed through flow cytometry (FACS).

In the spleen, 10 mg/kg of the test group mixed with lupeol and dowsterol 1:4 compared to the test group of 10 mg/kg/day administered with Lupeol or Daucosterol alone. It was confirmed that CD8+/CD69+, a marker of cytotoxic T cells, which are immune cells in the test group administered per day, increased to the highest level. As a result, it was confirmed that the 1:4 mixture of lufeol and docosterol showed synergy.

The results are shown in FIG. 2.

Experimental Example 3: Test of CD11c+/CD69+ cell level change in mesenteric lymph nodes by ingestion of Lupeol and Dacosterol in an inflammatory bowel disease mouse model (IBD, Inflammatory bowel disease)

In mice with inflammatory bowel disease (IBD), Lupeol and Daucosterol were administered at 10 mg/kg/day for each test group, and Lupeol and Daucosterol were 1:4. The mixed sample was orally administered to the control group at 10 mg/kg/day for 14 days daily. Dextran sodium sulfate (DSS) was administered orally on the 15th day to induce inflammatory bowel disease, and samples were administered to the mixed administration group of Lupeol, Daucosterol and Lupeol and Dowosterol 1:4 for 9 days, respectively. Was administered. After completion of the test, the immune organs of the IBD mouse were separated and the immune cell changes of each immune organ were measured.

In order to confirm the activity and number change in the cells of each immune organ isolated from the IBD mouse model, the antibodies against the cell surface molecules serving as the respective markers were reacted and analyzed through flow cytometry (FACS).

In mesenteric lymph nodes (MLN, Mesenteric lymph node), the test was a mixture of lupeol and docosterol in a ratio of 1:4 compared to the test group administered with 10 mg/kg/day of Lupeol or Daucosterol alone. In the test group administered with 10 mg/kg/day, it was confirmed that CD11c+/CD69+, which are markers such as macrophage (Macrophage), monocytes, dendritic cells, etc., which are immune cells, increased to the highest level. As a result, it was confirmed that the 1:4 mixture of lufeol and docosterol showed synergy.

The results are shown in FIG. 3.

Experimental Example 4: Test for measuring the level of decrease in the amount of free radicals that is increased when LPS (lipopolysaccharide) treatment in a mouse macrophage cell line (RAW264.7) is reduced by treatment with Lupeol and Dacosterol

When the mouse macrophage cell line RAW264.7 cells are treated with gram-negative bacterial cell wall component LPS (lipopolysaccharide), they generate free radicals (ROS) by an immune response, and when the test substance is treated, the amount of free radicals (ROS) decreases Therefore, the immune enhancing activity can be evaluated.

RAW264.7 cells, a mouse macrophage cell line, were dispensed into 12 well plates to be 10 ⁵ cells/well, and cultured for 24 hours. After treating the cultured cells with a concentration of 500 ng/ml LPS (lipopolysaccharide), the cells were cultured again for 24 hours to induce RAW264.7 cells to generate free radicals (ROS). The test substance, Lupeol, Daucosterol, and a mixture of Lufeol and Daucosterol 1:4 were treated at a concentration of 100 ug/ml, and then cultured for 6 hours. For the measurement of free radicals (ROS), 2',7'-dichlorofluorescin diacetate (DCF-DA; 20 μM) was treated to analyze fluorescently stained cells through flow cytometry (FACS).

Compared with the test group treated with Lupeol or Daucosterol alone, the highest free radical (ROS) reduction in the test group treated with the sample mixed with 1:4 of Lufeol and Dowsterol It was shown.

The results are shown in FIG. 4.

Formulation example:

[Production Example 1] Preparation of powder

20mg of a 1:4 mixture of lufeol and daucosterol

Lactose 100mg

Talc 10mg

The above ingredients are mixed and filled in an airtight fabric to prepare a powder.

[Formulation Example 2] Preparation of tablets

10 mg of a 1:4 mixture of lufeol and daucosterol

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 1 mg

Microcrystalline cellulose 84 mg

Povidone 3 mg

Sodium starch glycolate 2 mg

After mixing the above components, tablets are prepared by tableting according to a conventional tablet manufacturing method.

[Formulation Example 3] Preparation of healthy food

1000 mg of a 1:4 mixture of lufeol and daucosterol

Vitamin mixture

Vitamin A Acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ folic acid

Calcium Pantothenate 0.5 mg

Suitable amount of mineral mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium phosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the vitamin and mineral mixture is a composition suitable for a relatively healthy food in a preferred embodiment, the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for preparing a healthy food. , Granules can be prepared and used in the preparation of healthy food compositions according to conventional methods.

[Formulation Example 4] Preparation of healthy drinks

1000 mg of a 1:4 mixture of lufeol and daucosterol

Citric acid 1000 mg

Oligosaccharide 100 g

Taurine 1 g

900 ml total by adding purified water

After mixing the above components according to a conventional health drink manufacturing method, and stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered, obtained in a sterilized 2 liter container, sealed and sterilized, then stored in the refrigerator and then stored in the present invention. Used in the manufacture of healthy beverage compositions

Although the above composition ratio is a mixture of components suitable for a preference beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand nation, and usage.

Nr in the drawing is an abbreviation for Normal.
DA on the drawing is an abbreviation for Daucosterol.
WT on the drawing is an abbreviation for Wild type.

Claims (6)

A composition for enhancing immunity, comprising a mixture of lupeol and dacosterol as an active ingredient.
According to claim 1,
The mixture of the lupeol (Lupeol) and docosterol (Daucosterol) is a composition for enhancing immunity, characterized in that contained in 0.0001 to 50% by weight relative to 100% by weight of the total composition based on solids.
According to claim 1,
The lupeol (Lupeol) and docosterol (Daucosterol) is a composition for enhancing immunity, characterized in that it is included in a weight ratio of 1:1 to 1:10.
According to claim 1,
The immunity-enhancing composition is a tablet, pill, powder, granule, powder, liquid, spray, or capsules prepared for immunity.
Functional food comprising the composition for enhancing immunity according to any one of claims 1 to 4.
A pharmaceutical composition comprising the composition for enhancing immunity according to any one of claims 1 to 4.

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