KR101862282B1 - A Composition Comprising the combined herbal extract of Dendropanax morbifera and Acanthopanax senticosus HARMS for immuno-stimulating activity - Google Patents

Abstract

The present invention relates to a composition for prevention and treatment of immune enhancement or hypoimmunopathy comprising a complex herbal medicine extract comprising a mixture of a woody plant extract and an astringent plant, wherein the extract of the present invention is used in an experiment using an immunodeficient mouse, Effect on efficacy The effect on NK cell activity in the experiment The NK cell activity was measured in the experiment. As a result, the NK cell activity was higher in the group consumed with the mixture of Hwangchulchae and Rhizoma than in the comparative group consumed alone And T / B cell proliferative activity. In the experiment, the T / B proliferative capacity reduced to the nutritional deficiency was significantly increased through the ingestion of the mixture of Ganoderma lucidum and Ganoderma lucidum, In the cytokine secretion assay, cytokine (IL-2, IFN-γ) secretion was measured Visible kigo Th2 cytokine (IL-4, 6, 10, TNF-α) secretion and reduce In particular, it was confirmed that the mutant function was synergistically increased in terms of immune function and cytokine secretion ability than the comparative group in which each of the wild grasses and goatskin were individually ingested, thereby enhancing the immune function synergistically, thereby activating the immune function, A health food, a functional food, a health supplement, a food additive, and the like.

Description

[0001] The present invention relates to a composition for enhancing immunity comprising a herbal extract comprising a combination of a herbal extract and a herbal extract. [0002] The present invention relates to a composition for enhancing an immune response comprising Dendropanax morbifera and Acanthopanax senticosus HARMS for immuno-stimulating activity,

The present invention relates to an immunoconjugate composition containing a herbal extract comprising a combination of an extract of Ganoderma lucidum and Ganoderma lucidum.

[Literature 1] Immunology. 49, 1992; Dig dis Sci. 52, pp 1890-1896, 2007; J Life Sci. 19, pp 479-485, 2009; Biol Pharm Bull. 27, pp 617-620, 2004

[Document 2] J. Allergy clin immunol. 9, pp 616, 1993; Immunology. 47, pp 75, 1982

[Literature 3] A-Reum Yu, Ho-Young Park, Yun-Sook Kim, Sang-Keun Ha, Hee-Do Hong, Hee-Don Choi. 2012. Immuno-enhancing Effect of Seed Extracts on a RAW 264.7 Macrophage Cell Line. J Korean Soc Food Sci Nutr 41 (12): 1671-1676

[4] Uthaisangsook S, Day NK, Bahna SL, Good RA, Haraguchi S. 2002. Innate immunity and its role against infections. Ann. Allergy Asthma Immunol. 88: 253-264

[5] Birk RW, Gratchev A, Hakiy N, Politz O, Schledzewski K, Guillot P, Orfanos CE, Goerdt S. 2001. Alternative activation of antigen-presenting cells: concepts and clinical relevance. Hautarzt 52: 193-200

[6] Seon A Yoo, Ok Kyung Kim, Da-Eun Nam, Yongjae Kim, Humyoung Baek, Woojin Jun, Jeongmin Lee. 2014. Immunomodulatory Effects of Fermented Curcuma longa L. Extracts on RAW 264.7 Cells. J Korean Soc Food Sci Nutr 43 (2): 216-243

[Literature 7] Myung-Woo Byun. 2013. Immunomodulatory Activities of Apple Seed Extracts on Macrophage. J Korean Soc Food Sci Nutr 42 (9): 1513-1517

[Reference 8] Yi Seul Seo and Kwang-Soon Shin. 2012. Immune System-Stimulating Activities of Mucilage Polysaccharides Isolated from Opuntia humifusa. J Korean Soc Food Sci Nutr 41 (1): 95-102

[9] A-Reum Yu, Ho-Young Park, In-Wook Choi, Yong-Kon Park, Hee-Do Hong, Hee-Don Choi. 2012. Immune Enhancing Effect of Medicinal Herb Extracts on a RAW 264.7 Macrophage Cell Line. J Korean Soc Food Sci Nutr 41 (11): 1521-1527

[Literature 10] Journal of Ethnopharmacology 90, 403-408 (2004)

[Document 11] Korean Patent Registration No. 10-1194947

[Document 12] Korean Patent Registration No. 10-1443230

[Document 13] Korean Patent Laid-open Publication No. 10-2015-0056721

[Literature 14] Information Sourcing, Illustrated Dictionary of Contemporary Art, Yeongrim Publishers, pp432-435, 1998

[Literature 15] Information Sourcing, Illustrated Dictionary of Contemplation, Younglim Corporation, p858, 1998

The human body has the characteristic of trying to maintain homeostasis, and it also works in the immune system. Immunization means a series of bio-defense reactions that occur when a substance other than its own components breaks the homeostasis of the body or threatens itself. It protects the body from harmful substances entering through the skin, digestive tract, and respiratory system. This is a defense system that removes external stimulants, or acts to reduce serious inflammation in the wound. All immune responses, including the inflammatory response activated by influx of pathogens in the body, return to normal after removal of the pathogen, but when the immune response is difficult, immune responses are significantly higher or lower than those of normal individuals do. Immune deficiency or deficiency of the immune function is called the immune system in this state, the immune response is not activated properly, the body does not respond properly to foreign substances, causing the infection. In contrast, immune hypersensitivity is a condition in which some immune responses overreact, resulting in an imbalance of the immune system resulting in an allergic reaction (Immunology, 49, 1992; Dig dis Sci., 52, pp 1890-1896, 2007; J Life Sci. 19, pp 479-485, 2009; Biol Pharm Bull. 27, pp 617-620, 2004).

If the immune system is not properly regulated and becomes unbalanced, imbalance of cytokine in the body, unbalance of T / B cell proliferation, depletion of antioxidant nutrients may cause impaired immune regulation and increase inflammatory substance secretion, And it can not cope with influx of pathogens, so that inflammation can be intensified. Therefore, the immune response of the human body is balanced and the control can be maintained normally, so that the immune control ability is important for the prevention and treatment of disease (J Allergy Clin Immunol., 9, pp 616, 1993; ).

In the spleen, it is involved in immune regulation and secretes cytokines such as interferon (IFN) -γ, tumor necrosis factor (TNF) -α and interleukin (IL) -1β, IL-5, IL-6 and PGE2. These cytokines act in the execution phase of the immune response and play an important role in the signal transduction between the immune cells and the inflammatory system.

Since 2000, acute respiratory syndrome SARS, avian flu, Ebola virus, and other acute new infectious diseases have emerged worldwide, and the importance of immunity, which is the defense ability of the human body, is increasing. Immunity, which is a bio-defense against biological threats such as breakdown of homeostasis in the body or pathogen infection, is a specific immunity involving nonspecific immunity involving macrophages or natural killer (NK) cells and T cells or B cells (1. A-Reum Yu, Ho-Young Park, Yun-Sook Kim, Sang-Keun Ha, Hee-Do Hong, Hee-Don Choi. 2012. Immuno-enhancing Effect of Seed Extracts on a RAW 264.7 Macrophage Cell Line. J Korean Soc Food Sci Nutr 41 (12): 1671-1676). Phagocytes such as neutrophile, monocyte, and macrophage are the primary cells of the innate immune response that protects the human body from pathogens when pathogens enter the body (Uthaisangsook S, Day NK, Bahna SL, Good RA, Haraguchi S. 2002. Innate immunity and its role against infections. Ann. Allergy Asthma Immunol. 88: 253-264). In particular, macrophages function as antigen-presenting cells as the first counterparts in biological defense after epithelial cell barrier, and also affect T cells in response to adaptive immunity (3. Birk RW, Gratchev, Hakiy N, Politz O, Schledzewski K, Guillot P, Orfanos CE, Goerdt S. 2001. Alternative activation of antigen-presenting cells: concepts and clinical relevance. Hautarzt 52: 193-200) (TNF-α) and nitric oxide (NO), which contribute to the immune response, and secrete cytokines such as tumor necrosis factor-α (TNF-α) Humyoung Baek, Woojin Jun, Jeongmin Lee 2014. Immunomodulatory Effects of Fermented Curcuma longa L. Extracts on RAW 264.7 Cells. J Korean Soc Food Sci Nutr 43 (2): 216-243).

Research on natural substances to find the active factor of macrophages responsible for congenital immune function that can cope with diseases before adaptive immune function of human body is actively proceeded. 2012. Immune Enhancing Effect of Medicinal Herb Extracts on a RAW 264.7 Macrophage Cells (A-Reum Yu, Ho-Young Park, In-Wook Choi, Yong-Kon Park, Hee- In this study, we investigated the effect of the extract of apple juice on the immune response of macrophages using macrophage extracts. In the present study, we investigated the effect of fermented kojang on the immune response of RAW264.7 macrophages. Respectively. (4Seon A Yoo, Ok Kyung Kim, Da-Eun Nam, Yongjae Kim, Humyoung Baek, Woojin Jun, Jeongmin Lee. 2): 216-243)

       Macrophages are the phagocytic immune cells that play an important primary defense function for the immune immune system that exists in almost all tissues of the human body. Macrophage is an immune cell that secretes a number of cytokines and plays a pivotal role in the immune response to antigens in the process of detecting and removing germs and foreign substances (12Yi Seul Seo and Kwang-Soon Shin, 2012. Immune System-Stimulating Activities of Mucilage Polysaccharides Isolated from Opuntia humifusa. J Korean Soc Food Sci Nutr 41 (1): 95-102). The NO produced in macrophages is an important signaling agent that protects against tumor cells and infected microorganisms in the immune system. The effect of NO on RAW264.7 in herbal medicine extracts of licorice, (LPS), which is similar to the treatment of LPS, was shown to be effective for the production of NO in macrophages (5.A-Reum Yu, Ho-Young Park, Yong-Kon Park, Hee -Do Hong, Hee-Don Choi, 2012. Immune Enhancing Effect of Medicinal Herb Extracts on a RAW 264.7 Macrophage Cell Line. J Korean Soc Food Sci Nutr 41 (11): 1521-1527).

  Dendropanax morbifera is an evergreen broad-leaved arboreous tree native to Southwest Jeollanam-do and Jeju-do, which belongs to the Araliaceae of Korea. It is a species that does not fall into winter when it blooms in the winter. When it bruises the bark, yellow resinous juice comes out, ).

    Huangchil has been used as a rare paint that emits golden colors of emperor's armor, helmets and other metal ornaments since the Three Kingdoms period. It has been used in the Goryeo period, written in the Goryeo period, in the Guilin variety in China, , And it is said that it was a special product of Baekje before the time, and it remains in the book service area of the Tang Dynasty history book. Dendrofenacoids and polyacetylenic compounds have been found in the study of their constituents (Journal of Ethnopharmacology 90, 403-408 (2004) Park, Bo-Young, etc.) Considering the fact that it has been used in various diseases, there is almost no scientific identification of it.

        An invention relating to a cosmetic composition comprising an extract of Dendropanax morbifera as an active ingredient (Korean Patent No. 10-1176529); A composition for improving blood pressure, a composition for improving blood pressure, a composition for improving liver function (Korean Patent Registration No. 10-1443230), a composition for improving blood pressure, A hangover remover containing Hwangchujang and a method for producing the same (Korean Patent Laid-Open No. 10-2015-0056721).

Acanthopanax is a plant of the Araliaceae family, senticosus HARMS), the components include a large amount of glycoside, daucosterol, ethyl-alpha-galactoside, syringaresinol glucoside, syringin, And it is known that it is distributed throughout the country and exhibits adaptogens. (Information Sourcing, Illustrated Dictionary of Illustrations, Young Lim Publishing House, pp432-435, 1998).

Therefore, the inventors of the present invention measured weight and dietary efficiency in an experiment using an immunodeficient mouse of the present invention in an experiment on the effect of body weight and diet efficiency on experimental animals, was typically low, and the weight gain in the DM, aS, low, High group were significantly increased compared to the mal group, feed efficiency was confirmed that increased significantly compared to the mal group in DM, aS, High group (experimental Example 1-1-1 ); Effects on NK cell activity In the experiment, NK cell activity was found to increase synergistically with NK cell activity than the comparative group, which consumed a mixture of Hwaseum japonica and Caerimaceae, alone. It was to be (example 1-2- 1); The T / B cells decreased to malnutrition in the impact test on the proliferation T / B The proliferation was confirmed that there is a significant immunomodulatory effect in the increase in through the intake of hwangchil trees and gasiohgapi mixture (Example 1-3- 1 ); In a cytokine secretion measurement experiment, Th1 Increase the cytokine (cytokine) (IL-2, IFN-γ) secretion and Th2 cytokine (IL-4, 6, 10, TNF -α) reducing secretion and in particular, hwangchil wood, increases in the comparison group of cytokines associated with immune system than Kin secretion side for intake of the gasiohgapi alone synergistically synergistic immune ( Experimental Example 1-4-1 ) to activate the immune function, thereby being usefully used as a composition for the treatment and prevention of hypoimmune diseases, a health functional food, a functional food, a health supplement, a food additive, etc. The present invention has been completed.

In order to accomplish the above object, the present invention provides an immunoconjugate composition containing a complex herbal medicine extract consisting of Hwangbukchae and Rhizoma griseo.

In order to solve the above-mentioned problems, the present invention provides a pharmaceutical composition for preventing or treating immunity enhancement or hypoimmunities, which comprises an extract of a complex herb medicine comprising a mixture of woody spruce tree and scabbard as an active ingredient.

The herbal medicine extracts as defined herein preferably have a weight mixing ratio (w / w) of 1 to 100: 0.01 to 1 part by weight (w / w), more preferably 1-50: 0.1-1 (W / w), more preferably 1-30: 1 part by weight of the composition. The extracts defined herein include extracts which are soluble in water, water, alcohol, lower alcohols having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water or a mixed solvent of water and alcohol, including purified water.

The pharmaceutical composition of the extract may be used in an amount of 0.1 to 90% by weight based on the total weight of the composition.

The immunosuppressive conditions defined herein include those caused by a decrease in immune function by chemotherapy or radiation therapy or a disease caused by immune reduction after bone marrow transplantation, a disease caused by immune system damage and a cancer disease caused by a decrease in immune function, , Preferably a disease caused by a decrease in immune function by chemotherapy or radiotherapy or immunological degradation after bone marrow transplantation, more preferably a bacterial / viral infection in an elderly person, a chronic respiratory infection, a chronic urinary tract infection , Pressure sores, flu, pneumonia, varicella, measles, sudden rash, foot flu, rubella, or Crohn's disease.

Hereinafter, the present invention will be described in detail.

The extract of the present invention can be obtained as follows.

The weight ratio (w / w) of the whitish wood and the sea mustard is 1-100: 0.01-1 parts by weight (w / w), more preferably 1-50: 0.1 to 1 part by weight (w / w), and even more preferably 1-30: 1 part by weight to obtain a formulation sample; The sample of the first stage is washed and the sample is washed with water, methanol, ethanol or a mixed solvent thereof with a weight of 1 to 20 times (w / v), preferably 1 to 10 times (w / v) A cold extraction method or an ultrasonic extraction method in a mixed solvent of water and ethanol at 50 to 120 ° C, preferably about 80 to 100 ° C for 1 hour to 5 hours, preferably 2 hours to 4 hours, A second step of performing an extraction method; A third step of filtering the extract obtained in the above step with a filter paper to obtain a filtrate; The crude herbal medicine extract of the present invention can be prepared through a fourth step in which the filtrate is lyophilized, dried at room temperature or hot air dried, and preferably freeze-dried to obtain a dry herbal herb extract.

Accordingly, the present invention provides a pharmaceutical composition and a health functional food for the prevention and treatment of immune enhancement or immunosuppression, which comprises the herbal extract obtained from the above-described production method and the herbal extract of Hwasungi .

In the experiment using the immuno-deficient mice of the present invention prepared by the above-mentioned method, weight and dietary efficiency of the experimental animals were tested in terms of body weight change and diet efficiency. As a result, , And DM, AS, Low, and High groups showed significantly higher weight gain than the mal group, and DM, AS, and High groups showed a significant increase in body weight gain compared to the mal group ( Experimental Example 1-1-1 ); Effects on NK cell activity In the experiment, NK cell activity was found to increase synergistically with NK cell activity than the comparative group, which consumed a mixture of Hwaseum japonica and Caerimaceae, alone. It was to be (example 1-2- 1); The T / B cells decreased to malnutrition in the impact test on the proliferation T / B The proliferation was confirmed that there is a significant immunomodulatory effect in the increase in through the intake of hwangchil trees and gasiohgapi mixture (Example 1-3- 1 ); In a cytokine secretion measurement experiment, Th1 Increase the cytokine (cytokine) (IL-2, IFN-γ) secretion and Th2 cytokine (IL-4, 6, 10, TNF -α) reducing secretion and in particular, hwangchil wood, increases in the comparison group of cytokines associated with immune system than Kin secretion side for intake of the gasiohgapi alone synergistically synergistic immune ( Experimental Example 1-4-1 ) to activate the immune function, thereby being usefully used as a composition for the treatment and prevention of hypoimmune diseases, a health functional food, a functional food, a health supplement, a food additive, etc. And it was confirmed to be useful for the treatment and prevention of hypoimmunities.

Accordingly, the present invention provides an immuno-enhancement or hypoimmune pharmaceutical composition comprising the herbal extract of Hwangchujanggul and the herbal composition as an active ingredient.

The composition for the prevention and treatment of hypomagnesa of the present invention contains 0.1 to 99% by weight of the above extract relative to the total weight of the composition.

However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.

The composition comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.

The composition containing the extract according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like in the form of oral preparations or external preparations, Examples of carriers, excipients and diluents that can be included in the composition containing the extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . The external preparation for parenteral administration includes sterilized aqueous solution, non-aqueous solution, suspension, emulsion, freeze-dried preparation, and suppository. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The preferred dosage of the extract of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the administration route and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at a dose of 0.01 mg / kg to 10 g / kg per day, preferably 1 g / kg to 5 g / kg per day. The administration may be carried out once a day or divided into several doses. Accordingly, the dosage is not limited in any way to the scope of the present invention.

The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

In addition, the present invention provides a health functional food for preventing or ameliorating immune enhancement or hypoimmunity, which contains the herbal extract of the present invention comprising the combination herbal extracts comprising the above-mentioned Yellowtail and woody vegetables.

The present invention also provides a health supplement for preventing or ameliorating immune enhancement or hypoimmunity, which comprises the herbal extract of the present invention comprising the combination herbal extracts comprising the above-mentioned Hwangchujanggwa and Ganoderma lucidum.

Accordingly, the present invention also provides a food or food additive for preventing or improving immunity enhancement or hypoimmunity, which comprises an extract of a complex herb medicine comprising an extract of Ganoderma lucidum and Ganoderma lucidum as an active ingredient.

The composition containing the extract of the present invention can be used variously for medicines, foods and beverages for the prevention and improvement of hypoimmunities. Examples of foods to which the extract of the present invention can be added include various kinds of foods such as candy, beverages, gums, tea, vitamin complex, health supplement foods and the like, and they are in the form of powder, granule, tablet, capsule or beverage Can be used.

Since the extract of the present invention has little toxicity and side effects, it can be safely used even for long-term administration for preventive purpose.

The extract of the present invention can be added to food or beverage for the purpose of prevention and improvement of hypoimmunities. At this time, the amount of the extract in the food or beverage is generally 0.01 to 15% by weight of the total food weight of the health food composition of the present invention, and 0.02 to 10 g, Can be added in a proportion of 0.3 to 1 g.

The food of the present invention may be a health supplement food, a health functional food, a functional food, and the like, but is not limited thereto and includes the addition of the extract of the present invention to natural foods, processed foods, patient foods, and general food materials. The food composition of the present invention can be used as it is or in combination with other food or food compositions, and can be suitably used according to conventional methods. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, improvement, or therapeutic treatment). In general, the extract of the present invention may be added in an amount of 0.1 to 70 parts by weight, preferably 2 to 50 parts by weight, based on 100 parts by weight of the raw material for food or beverage in the production of food or beverage. The effective dose of the above extract may be used in accordance with the effective dose of the pharmaceutical composition, but may be less than the above range for health and hygiene purposes or long-term intake for health control purposes, It can be used in an amount exceeding the range described above. There is no particular limitation on the kind of the food. The food composition may be used in the form of tablets, hard or soft capsules, liquids, suspensions, and the like, which may contain conventional excipients, such as excipients in the case of oral preparations, Binders, disintegrators, lubricants, solubilizers, suspending agents, preservatives or extenders.

Examples of the food to which the above extract can be added include dairy products such as meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen noodle, gums, ice cream, various soups, drinks, tea, Alcoholic beverages and vitamins complex, and other nutrients, but the present invention is not limited to these kinds of foods.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient, as long as it contains the extract as an essential ingredient at the indicated ratio, and there is no particular limitation to the liquid ingredient. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like Sugar, and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

As described above, in the experiment using the immunodeficient mouse of the present invention, the body weight and the diet efficiency were measured in the experiment of the effect on the body weight change and the diet efficiency of the experimental animals. As a result, . In DM, AS, Low and High groups, weight gain was significantly higher than that of mal group. Dietary efficiency was significantly increased in DM, AS and High group compared to mal group 1-1-1); Effects on NK cell activity In the experiment, NK cell activity was found to increase synergistically with NK cell activity than the comparative group, which consumed a mixture of Hwaseum japonica and Caerimaceae, alone. (Experimental Example 1-2-1); Effect on T / B Cell Proliferation Ability In the experiment, it was confirmed that the T / B proliferative capacity reduced to the nutritional deficiency was significantly increased through the ingestion of the mixture of Ganoderma lucidum and Ganoderma lucidum, One); In the cytokine secretion assay, the secretion of Th1 cytokines (IL-2, IFN-γ) and the secretion of Th2 cytokines (IL-4, 6, 10 and TNF-α) (1), (2), (3), (4), (5), (6), (7), (7), and (7) of the present invention, the immune function , Drugs for the treatment and prevention of hypoimmunities, health functional foods, functional foods, health supplements, food additives and the like.

FIG. 1 is a graph showing the cytotoxicity of NK (Natural Killer) cells to Yac-1 cells obtained from a spleen in a C57BL / 6 mouse experimental model (NK cells (110 ⁵ cells / well) Cells were coincubated with each other and incubated for 4 hours. Cell activity was measured with an ELISA reader. All data were expressed as mean ± standard deviation (SD) Six times (n = 6), NC: Normal control, Mal: Malnutrition, DM: Dendropanax Morbifera L. 300 mg / kg BW, AS: Acantho Panax senticosus 300 mg / kg BW, Low: DM + AS 100 mg / kg BW, and High: DM + AS 300 mg / kg BW, respectively);
FIG. 2 is a graph showing the effect of the C57BL / 6 mouse experimental model on the T / B cell proliferation obtained from the spleen (after 48 hours of culture, the proliferation was measured by an ELISA reader, Normal control, Mal: Malnutrition, DM: Dendropanax (n = 6), NC: Normal control, Morbifera L. 300 mg / kg BW, AS: Acantho Panax senticosus 300 mg / kg BW, Low: DM + AS 100 mg / kg BW, and High: DM + AS 300 mg / kg BW, respectively);
FIG. 3 is a graph showing the effect of the cytokines (IL-2, IFN-γ) obtained from the spleen on the C57BL / 6 mouse experimental model (cytokines were measured by an ELISA reader, (n = 6), NC: normal control, Mal: malnutrition, DM: Dendropanax Morbifera L. 300 mg / kg BW, AS: Acantho Panax senticosus 300 mg / kg BW, Low: DM + AS 100 mg / kg BW, and High: DM + AS 300 mg / kg BW, respectively);
FIG. 4 is a graph showing the effect of C57BL / 6 mouse test sample on the cytokines (IL-4, IL-6) obtained from the spleen (cytokines were measured by an ELISA reader, (n = 6), NC: normal control, Mal: malnutrition, DM: Dendropanax Morbifera L. 300 mg / kg BW, AS: Acantho Panax senticosus 300 mg / kg BW, Low: DM + AS 100 mg / kg BW, and High: DM + AS 300 mg / kg BW, respectively);
FIG. 5 is a graph showing the effect of the C57BL / 6 mouse sample on the Th2 cytokines (IL-10, TNF-α) obtained from the spleen in the experimental model. (N = 6), NC: normal control (SD), and each experiment was performed 6 times (n = 6) Mal: Malnutrition, DM: Dendropanax Morbifera L. 300 mg / kg BW, AS: Acantho Panax senticosus 300 mg / kg BW, Low: DM + AS 100 mg / kg BW, and High: DM + AS 300 mg / kg BW, respectively).

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

COMPARATIVE EXAMPLE 1. Preparation of extract of Hwangchil (comparative)

(V / w) of water was added to 500 g of Hwangchu-myeon, which is cultivated in Cheju-do farm, of Hambakbang BioPharm Co., Ltd. for 5 hours by reflux extraction at 90 ℃ or higher. The extract was filtered with a filter paper (Whatman), and then extracted with water extract (hereinafter referred to as DM) by Eyela (Rotary Evaporator N-1000) and freeze-drying (Eyela, Freeze Dryer FDU-540) Was used as a comparative sample in the following experimental examples.

COMPARATIVE EXAMPLE 2 Preparation of Acanthopanax senticosus extract (for comparison)

Water (v / w) was added to 500 g of cauliflower grown in Nonsan Farm, Chungnam Province and extracted with reflux extraction for 5 hours at 90 ℃ or higher. The extract was filtered with a filter paper (Whatman), and then concentrated under reduced pressure (Eyela, Rotary Evaporator N-1000) and freeze-dried (Eyela, Freeze Dryer FDU-540) to obtain a watery extract (hereinafter referred to as AS) Was used as a comparative sample in the experimental example.

Example 1: Preparation of mixed extract of Hwangchujanggwa and Ganoderma lucidum

(V / v) was added to 500 g of a mixture of Hwangchulchae grown on a Jeju Island farm and a mixture of Kwachyogi cultivated on a Nonsan farm in Chungcheongnam-do, at a ratio of 7: 3 (weight ratio) w) and extracted by reflux extraction at a temperature of 90 ° C or higher for 5 hours. The extract was filtered with a filter paper (Whatman), and then the complex herbal medicine water extract (hereinafter referred to as DMS) was obtained by concentration under reduced pressure (Eyela, Rotary Evaporator N-1000) and freeze drying (Eyela, Freeze Dryer FDU-540) Was used as a sample in the following experimental example.

Experimental Example  1. 7.5% 미ron 유통  (Vitamin, Mineral) deficiency in the mouse model of immunodeficiency mice. in vivo  Immunomodulatory efficacy experiment

1-1. Preparation and treatment of experimental animals

1-1-1. Animal processing

: C57BL / 6 mouse mice (Central experimental animals) were supplied with 4 weeks old female mice of about 20 g / day and light and dark cycles were performed at a temperature of 23 ± 2 ° C and a relative humidity of 50 ± 5% Were used for the experiment through the adaptation device for one week. During the adaptation period, AIN-93G diet and drinking water were allowed to be freely consumed, and the animals determined to be healthy during the adaptation period were measured for body weight, and individuals close to the average weight were selected and grouped by randomization.

1-1-2. Dietary Preparation and Test Group Separation

: A mouse with a 7.5% micronutrient (vitamin, mineral) deficiency disrupts the immune system for 10 weeks to produce an immunocompromising animal model. The water extract of Huangchuang, water extract of Ganoderma lucidum and mixture of Ganoderma lucidum and Ganoderma lucidum (100 mg / kg bw And 300 mg / kg bw), diets and body weights were measured twice weekly and sacrificed 10 weeks later. The test groups were as follows: normal group (NC), control group (Mal), yellowtail water extract treatment group (DM), astringent water extract treatment group (AS) (High) treatment group (Table 1).

Table 1. Experimental design of animal (n = 6 / group) Group Immune deficiency Provided diet NC - AIN 93G Mal + 7.5% micronutrient in AIN 93G DM + DM 300 mg / kg B.W. in 7.5% micronutrient AIN 93G AS + AS 300 mg / kg B.W. in 7.5% micronutrient AIN 93G Low + DM + AS 100 mg / kg B.W. in 7.5% micronutrient AIN 93G High + DM + AS 300 mg / kg B.W. in 7.5% micronutrient AIN 93G

1-2. Experimental course

: In order to confirm the effect of the sample of the above example on the immunoregulatory ability of the immunodeficient mouse , the following experiment was conducted by applying the method described below.

1-2-1. Effect of Mixed Extracts of Wood and Pine Needle on the Weight Change and Dietary Efficacy of Laboratory Animals

: The dietary weight and body weight were measured twice a week, and the body weight change and the dietary efficiency were calculated using the following equations (1) and (2).

1-2-2. Serum and tissue processing

: Experimental animals were fasted for 12 hours and anesthetized and laparotomized. The organs (spleen, liver, lymph nodes) were excised and weighed. The spleen was washed with Roswell Park Memorial Institute medium 1640 (Hyclone Laboratories, Logan, Utah, USA) immediately after extraction and the cell suspension obtained using a 0.45 μm cell strainer (BD biosciences, San Diego, USA) Cell lysing buffer (Sigma-Aldrich Co., St. Louis, Mo., USA) was used to hemolyze red blood cells. Separated spleen cells were used for cytotoxic and T and B cell proliferative activity, and isolated peritoneal macrophages were used for macrophage activation assay.

1-2-3. The extract of Hwangchu-gil In proliferation  Impact

: Splenocytes were plated at a density of 1 × 10 ⁶ cells / well in a 96-well plate, and 5 μg / mL of ConA (Sigma-Aldrich Co., St. Louis, MO, USA) Mitogen LPS (Sigma-Aldrich Co., St. Louis, Mo., USA) was treated at 5 μg / mL and cultured at 37 ° C and 5% CO ₂ . After 48 hours, 20 μL of Ez-Cytox (Dugen, Seoul, Korea) solution was added and absorbance was measured at 450 nm using an ELISA reader (VERSAMAXSL-20 Molecular Devices, Korea).

1-2-4. Cytokine ( Cytokine ) Secretion measurement

: Separated splenocytes were plated in 96 well plates at 1 × 10 ⁶ cells / well per well. IL-2, IL-4, IL-10 and IFN-γ were treated with 5 μg / mL of ConA and IL-6 and TNF-α were treated with 5 μg / mL of LPS. The supernatant was collected 24 hours after IFN-γ and 72 hours after IFN-γ and cultured at 37 ° C and 5% CO ₂ . Absorbance was measured at 650 nm with an ELISA reader using R & D system, USA.

1-3. Experiment result

1-3-1. The extracts of Hwangchujang and Ganoderma lucidum were compared with those of experimental animals Eating efficiency  Impact

The results of the experiment on the effect of nutrient deficiency on the body weight change and the diet efficiency of the mixed immuno-asymmetry models of the mixed extract of Ganoderma lucidum and Ganoderma lucidum var. .

Table 2. Mice body weight gain, food consumption and food efficiency rates of exprimental groups for 10 weeks. (n = 6 / group) Group ^{1 )} Weight gain ^{2 )} (g) Food consumption ^{3 )} (g) FER4 ⁾ NC 2.48 ± 0.79 ^a 1.83 ± 0.19 ^{ns 5)} 3.86 ± 1.27 ^a Mal -1.25 ± 2.3 ^c 1.73 ± 0.39 -3.96 + 3.11 ^c DM -0.05 ± 0.49 ^b 1.53 + - 0.27 -0.09 ± 0.92 ^b AS -0.28 ± 2.04 ^b 1.55 ± 0.3 -0.52 ± 3.77 ^b Low -0.75 + 0.08 ^b 1.48 0.18 -1.45 + 1.55 ^bc High 1.47 ± 0.32 ^a 1.61 + 0.19 2.61 ± 0.57 ^a Values are presented as means ± SD. Different letters show a significant difference at p <0.05 as determined by Duncan's multiple range test.
¹⁾ NC: Normal control, Mal: Malnutrition, DM: Dendropanax Morbifera L. 300 mg / kg BW, AS: Acantho Panax senticosus 300 mg / kg BW, Low: DM + AS 100 mg / kg BW, High: DM + AS 300 mg / kg BW
²⁾ Weight gain (g / 10weeks) = final body weight (g) initial body weight (g)
³⁾ Food consumption (g / day) = food consumption (g / day)
⁴⁾ Food efficiency rate (FER) = {weight (g) / food intake (g)} * 100
⁵⁾ ns: no significant difference

The body weight and dietary efficiency were significantly lower in the Mal group than in the NC group, and the weight gain was significantly increased in the DM, AS, Low and High groups compared to the mal group, , AS, and High groups, respectively.

1-3-2. The extracts of Hwangchu - NK  Effect on cell activity

FIG. 1 shows the results of an experiment on the NK cell activity of immunoequilibrium model animals with nutritional deficiency during 10 weeks of the above experiment for 10 weeks.

NK cell activity was decreased by 61.47% in the Mal group compared with the NC group. The NK cell activity of the sample group (DM, AS, Low, High) was 187.52% , 192.16%, 218.45%, and 233.69%, respectively. Especially, the group ingesting the mixed extract of Hwangchujanggwa and Kjelliphora increased the NK cell activity synergistically (hwangchil wood water extract-treated group (DM): 187.52%, gasiohgapi water extract-treated group (AS): 192.16%, hwangchil wood and gasiohgapi mixed extract low concentration treatment group (Low): 218.45%, hwangchil wood and gasiohgapi mixed extract a high concentration treatment (High: 233.69%) . Therefore, it was confirmed that it is effective for immunity enhancement.

1-3-3. The extracts of Hwangchujang and Ganoderma lucidum were used as T / B cells In proliferation  Impact

The results of the experiment on the effect of the mixed extract of Ganoderma lucidum and Ganoderma Lucidum on the T / B cell proliferation of the immuno-imbalance model animal model due to nutritional deficiency for 10 weeks were shown in FIG. 2 .

The T cell proliferative capacity was significantly decreased by 32.93% in Mal group compared to NC group. B cell proliferative capacity was significantly decreased by 30.26% in Mal group compared to NC group. T cell proliferation by T Cell proliferation was increased by 107.16%, 105%, 116.13% and 121.83%, respectively, in the sample intake (DM, AS, Low, High) In group 1, the T cell proliferative capacity was significantly increased as compared with that of Mal group. The B cell proliferative capacity of the sample was measured by the sample intake (DM, AS, Low, High) , 104.30%, 104.09%, 131.03%, and 131.03%, respectively. In particular, in the group fed with the mixed extract of Ganoderma lucidum and Ganoderma lucidum, the B cell proliferative capacity was significantly increased as compared with the Mal group, Group for comparison It was confirmed that the proliferation of B cells is confirmed to be increased synergistically.

 Therefore, it was confirmed that the T / B proliferative capacity reduced by nutritional deficiency was significantly increased through ingestion of the mixed extract of Ganoderma lucidum var.

1-3-4. Cytokine ( Cytokine ) Secretion measurement

(IL-2, IFN-γ) secretion in the experimental animal model of immune imbalance due to nutritional deficiency during 10 weeks of the experiment, The results of the experiment are shown in FIG.

IL-2 secretion was 6.56 ± 3.85 pg / mL in the Mal group and 117.67 ± 13.33 and 118.78 ± 7.70 in the sample intake (DM, AS, Low, High) , 154.33 ± 3.33, and 238.50 ± 29.17 pg / mL, respectively, which were significantly higher than those of Mal group,

- IFN-γ secretion was 297.22 ± 9.62 pg / mL in the Mal group and significantly lower in the NC group (1458.33 ± 83.33 pg / mL) than in the NC group (DM, AS, The IFN-γ secretion potency was 769.44 ± 38.49, 908.33 ± 83.33, 830.56 ± 19.25, and 730.56 ± 50.92 pg / mL, respectively, which were increased by 258.88%, 305.61%, 279.44%, and 245.79% It was confirmed that Th1 cytokine secretion concentration was synergistically increased compared with the comparative group ingested alone.

(IL-4, 6, 10, TNF-α) secretion in the mouse model of immune imbalance due to nutritional deficiency during 10 weeks of the experiment. The results of the experiment were shown in Figs. 4 to 5.

First, the IL-4 secretion level was 286.00 ± 8.66 pg / mL in the Mal group and 252.67 ± 2.89 and 269.33 in the sample intake (DM, AS, Low, High) ± 2.89, 259.33 ± 2.89, and 249.33 ± 2.89 pg / mL, respectively.

Serum IL-6 levels were significantly increased in the Mal group (393.50 ± 25.00 pg / mL) compared to the NC group (35.17 ± 2.89 pg / mL) The IL-6 secretion potencies were significantly reduced by 11.01%, 12.07%, 32.72% and 40.66%, respectively, compared with that of the Mal group, which was 350.17 ± 16.07, 346.00 ± 12.50, 264.75 ± 23.75 and 233.50 ± 15.00 pg /

IL-10 secretion was significantly increased in the Mal group (568.00 ± 52.92 pg / mL) compared to the NC group (354.67 ± 23.09 pg / mL) The IL-10 secretion potencies were significantly reduced by 23.47%, 21.13%, 32.86%, and 37.56%, respectively, compared with the Mal group, which was 434.67 ± 23.09, 448.00 ± 40.00, 381.33 ± 30.55 and 654.67 ± 23.09 pg / And Caesia cocinum were not significantly different from those of NC group.

The TNF-α secretion activity was significantly increased in the Mal group (440.56 ± 25.46 pg / mL) compared to the NC group (129.44 ± 19.25 pg / mL) The TNF-α secretion potency was 326.67 ± 8.33, 335.00 ± 16.67, 318.33 ± 16.67, and 189.17 ± 20.83 pg / mL, respectively, which were significantly lower than those of Mal group by 25.85%, 23.96%, 27.74% and 57.06%, respectively.

Statistical processing

Statistical analysis was performed using the one-way ANOVA test (Statistical Package for Social Science, version 20.0, SPSS Inc., Chicago, IL, USA). Statistical analysis was performed using a one-way ANOVA test The significance of the experimental groups was compared at Duncan's multiple range test at P <0.05 level.

The formulation examples of the compositions containing the extracts of the present invention are illustrated, but the present invention is not intended to be limited thereto but is only specifically described.

Formulation example  One. Sanje  Produce

DMS ---------------------------------------------- 20 mg

Lactose -------------------------------------------- 100 mg

Talc --------------------------------------------- 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2. Preparation of tablets

DMS ---------------------------------------------- 10 mg

Corn starch -------------------------------------- 100 mg

Lactose -------------------------------------------- 100 mg

Magnesium stearate ------------------------------- 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation example  3. Preparation of capsules

DMS --------------------------------------------- 10 mg

Crystalline cellulose - 3 mg

Lactose --------------------------------------- 14.8 mg

Magnesium stearate - 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

DMS --------------------------------------------- 10 mg

Mannitol ------------------------------------------ 180 mg

Sterile sterilized distilled water for injection - 297 mg

Na ₂ HPO ₄ 12H ₂ O -------------------------------------- 26 mg

(2 ml) per ampoule in accordance with the usual injection method.

Formulation example  5. Liquid  Produce

DMS --------------------------------------------- 20 mg

Isomerization Party ------------------------------------------ 10 g

Mannitol --------------------------------------------- 5 g

Purified water --------------------------------------------

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6. Of health food  Produce

DMS -------------------------------------------- 1000 mg

Vitamin mixture -------------------------------------

Vitamin A Acetate ------------------------------ 70 g

Vitamin E ---------------------------------------- 1.0 mg

Vitamin B ₁ --------------------------------------- 0.13 mg

Vitamin B ₂ --------------------------------------- 0.15 mg

Vitamin B ₆ ---------------------------------------- 0.5 mg

Vitamin B ₁₂ --------------------------------------- 0.2 g

Vitamin C ----------------------------------------- 10 mg

Biotin ------------------------------------------- 10 [mu] g

Nicotinic acid amide 1.7 mg

Folic acid --------------------------------------------- 50 μg

Calcium pantothenate ----------------------------------- 0.5 mg

Inorganic mixture -------------------------------------

Ferrous sulfate -------------------------------------- 1.75 mg

Zinc oxide --------------------------------------- 0.82 mg

Magnesium carbonate - 25.3 mg

Potassium phosphate monohydrate 15 mg

Secondary calcium phosphate -------------------------------------- 55 mg

Potassium citrate --------------------------------------- 90 mg

Calcium carbonate ---------------------------------------- 100 mg

Magnesium chloride ----------------------------------- 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation example  7. Manufacture of health drinks

DMS -------------------------- == ----------------- 100 mg

Vitamin C ------------------------------------------- 15 g

Vitamin E (powder) ------------------------------------ 100 g

Lactic Acid Iron ------------------------------------------ 19.75 g

Zinc oxide ------------------------------------------ 3.5 g

Nicotinic acid amide 3.5 g

Vitamin A ------------------------------------------ 0.2 g

Vitamin B ₁ ----------------------------------------- 0.25 g

Vitamin B ₂ ------------------------------------------ 0.3 g

Water ------------------------------------------------- - Quantity

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Formulation example  8. Cereal  Produce

DMS ---------------------------------------------- 7.51 g

Rice ---------------------------------------------- 40.49 g

Brown rice -------------------------------------------- 24.29 g

Sorghum --------------------------------------------- 2.02 g

Barley --------------------------------------------- 2.02 g

Glutinous rice --------------------------------------------- 2.02 g

Salt --------------------------------------------- 0.49 g

Ses ----------------------------------------------- 0.16 g

Sugar -------------------------------------------- 17.75 g

Sunflower oil --------------------------------------- 1.21 g

Water ------------------------------------------------- dose

The ingredients are mixed according to a conventional cereal manufacturing method and then used in the production of the cereal composition of the present invention after extrusion molding, primary drying, aging, ripening, syrup coating, secondary drying, and packaging.

Although the compositional ratio is a mixture of the components suitable for the preference, it is also possible to arbitrarily modify the blending ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Formulation example 9. Choco Crunch  Produce

DMS ------------------------------------------------- --- 5.00 g

Rice ------------------------------------------------- --- 21.50 g

Congratulations ------------------------------------------------ 23.02 g

Mixed cell milk powder ------------------------------------------ 27.78 g

Cocoa Butter -------------------------------------------- 15.35 g

Vegetable hardening oil ------------------------------------------ 13.02 g

Lecithin ------------------------------------------------- 0.15 g

Polyglycerin condensed ricinoleic acid ester 0.15 g

Vanillin ------------------------------------------------- 0.03 g

Formulation Example 10 . Manufacture of confectionery

DMS ------------------------------------------------- ---- 5.00 g

Starch ------------------------------------------------- --- 83.00 g

Rice flour ------------------------------------------------- 10.00 g

Sun salt ------------------------------------------------- - 0.5 g

Yuzu seasoning ----------------------------------------------- 1.5 g

Claims (8)

Immunostimulation or chemotherapy comprising a water extract of a combination herb comprising a mixture of woodwort and bitter gourd mixed with a weight ratio (w / w) of a woody bitter gourd and a bitter gourd (w / w) of 1-50: 0.1-1 parts by weight A pharmaceutical composition for the prevention and treatment of diseases with reduced immune function by chemotherapy such as radiation therapy. delete delete delete Immunostimulation or chemotherapy comprising a water extract of a combination herb comprising a mixture of woodwort and bitter gourd mixed with a weight ratio (w / w) of a woody bitter gourd and a bitter gourd (w / w) of 1-50: 0.1-1 parts by weight A health functional food for the prevention and improvement of diseases with reduced immune function by chemotherapy such as radiation therapy. The health functional food according to claim 5, which is in the form of a powder, a granule, a tablet, a capsule or a drink. Immunostimulation or chemotherapy comprising a water extract of a combination herb comprising a mixture of woodwort and bitter gourd mixed with a weight ratio (w / w) of a woody bitter gourd and a bitter gourd (w / w) of 1-50: 0.1-1 parts by weight Health supplements for prevention and improvement of diseases with lowered immune function by chemotherapy such as radiation therapy. delete

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