WO2024128690A1 - 면역저하상태에서의 인삼 열매 다당류의 선천성 면역조절 효과 - Google Patents
면역저하상태에서의 인삼 열매 다당류의 선천성 면역조절 효과 Download PDFInfo
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- WO2024128690A1 WO2024128690A1 PCT/KR2023/020172 KR2023020172W WO2024128690A1 WO 2024128690 A1 WO2024128690 A1 WO 2024128690A1 KR 2023020172 W KR2023020172 W KR 2023020172W WO 2024128690 A1 WO2024128690 A1 WO 2024128690A1
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- korean ginseng
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- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
- ZUFONQSOSYEWCN-UHFFFAOYSA-M sodium;2-(methylamino)acetate Chemical compound [Na+].CNCC([O-])=O ZUFONQSOSYEWCN-UHFFFAOYSA-M 0.000 description 1
- HYRLWUFWDYFEES-UHFFFAOYSA-M sodium;2-oxopyrrolidine-1-carboxylate Chemical compound [Na+].[O-]C(=O)N1CCCC1=O HYRLWUFWDYFEES-UHFFFAOYSA-M 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
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- SFVFIFLLYFPGHH-UHFFFAOYSA-M stearalkonium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCCCC[N+](C)(C)CC1=CC=CC=C1 SFVFIFLLYFPGHH-UHFFFAOYSA-M 0.000 description 1
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- 229960005322 streptomycin Drugs 0.000 description 1
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- 125000000020 sulfo group Chemical group O=S(=O)([*])O[H] 0.000 description 1
- AGGIJOLULBJGTQ-UHFFFAOYSA-N sulfoacetic acid Chemical compound OC(=O)CS(O)(=O)=O AGGIJOLULBJGTQ-UHFFFAOYSA-N 0.000 description 1
- 229940117986 sulfobetaine Drugs 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
- 239000002600 sunflower oil Substances 0.000 description 1
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- BORJONZPSTVSFP-UHFFFAOYSA-N tetradecyl 2-hydroxypropanoate Chemical compound CCCCCCCCCCCCCCOC(=O)C(C)O BORJONZPSTVSFP-UHFFFAOYSA-N 0.000 description 1
- DZKXJUASMGQEMA-UHFFFAOYSA-N tetradecyl tetradecanoate Chemical compound CCCCCCCCCCCCCCOC(=O)CCCCCCCCCCCCC DZKXJUASMGQEMA-UHFFFAOYSA-N 0.000 description 1
- 239000000892 thaumatin Substances 0.000 description 1
- 235000010436 thaumatin Nutrition 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 239000011747 thiamine hydrochloride Substances 0.000 description 1
- 235000019190 thiamine hydrochloride Nutrition 0.000 description 1
- 229960000344 thiamine hydrochloride Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000010936 titanium Substances 0.000 description 1
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- OGIDPMRJRNCKJF-UHFFFAOYSA-N titanium oxide Inorganic materials [Ti]=O OGIDPMRJRNCKJF-UHFFFAOYSA-N 0.000 description 1
- 229940042585 tocopherol acetate Drugs 0.000 description 1
- 235000015961 tonic Nutrition 0.000 description 1
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- 229960000716 tonics Drugs 0.000 description 1
- 235000010487 tragacanth Nutrition 0.000 description 1
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- LOIYMIARKYCTBW-OWOJBTEDSA-N trans-urocanic acid Chemical compound OC(=O)\C=C\C1=CNC=N1 LOIYMIARKYCTBW-OWOJBTEDSA-N 0.000 description 1
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- 235000013337 tricalcium citrate Nutrition 0.000 description 1
- 229960003500 triclosan Drugs 0.000 description 1
- WEAPVABOECTMGR-UHFFFAOYSA-N triethyl 2-acetyloxypropane-1,2,3-tricarboxylate Chemical compound CCOC(=O)CC(C(=O)OCC)(OC(C)=O)CC(=O)OCC WEAPVABOECTMGR-UHFFFAOYSA-N 0.000 description 1
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- 235000013769 triethyl citrate Nutrition 0.000 description 1
- SZEMGTQCPRNXEG-UHFFFAOYSA-M trimethyl(octadecyl)azanium;bromide Chemical compound [Br-].CCCCCCCCCCCCCCCCCC[N+](C)(C)C SZEMGTQCPRNXEG-UHFFFAOYSA-M 0.000 description 1
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- VLPFTAMPNXLGLX-UHFFFAOYSA-N trioctanoin Chemical compound CCCCCCCC(=O)OCC(OC(=O)CCCCCCC)COC(=O)CCCCCCC VLPFTAMPNXLGLX-UHFFFAOYSA-N 0.000 description 1
- APVVRLGIFCYZHJ-UHFFFAOYSA-N trioctyl 2-hydroxypropane-1,2,3-tricarboxylate Chemical compound CCCCCCCCOC(=O)CC(O)(C(=O)OCCCCCCCC)CC(=O)OCCCCCCCC APVVRLGIFCYZHJ-UHFFFAOYSA-N 0.000 description 1
- COXJMKGEQAWXNP-UHFFFAOYSA-N tris(14-methylpentadecyl) 2-hydroxypropane-1,2,3-tricarboxylate Chemical compound CC(C)CCCCCCCCCCCCCOC(=O)CC(O)(C(=O)OCCCCCCCCCCCCCC(C)C)CC(=O)OCCCCCCCCCCCCCC(C)C COXJMKGEQAWXNP-UHFFFAOYSA-N 0.000 description 1
- RHNXTZDKMRCKKT-UHFFFAOYSA-N tris(6-methylheptyl) 2-hydroxypropane-1,2,3-tricarboxylate Chemical compound CC(C)CCCCCOC(=O)CC(O)(C(=O)OCCCCCC(C)C)CC(=O)OCCCCCC(C)C RHNXTZDKMRCKKT-UHFFFAOYSA-N 0.000 description 1
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- 239000004474 valine Substances 0.000 description 1
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- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
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- 235000019155 vitamin A Nutrition 0.000 description 1
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- NCYCYZXNIZJOKI-UHFFFAOYSA-N vitamin A aldehyde Natural products O=CC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-UHFFFAOYSA-N 0.000 description 1
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- MECHNRXZTMCUDQ-RKHKHRCZSA-N vitamin D2 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)/C=C/[C@H](C)C(C)C)=C\C=C1\C[C@@H](O)CCC1=C MECHNRXZTMCUDQ-RKHKHRCZSA-N 0.000 description 1
- 235000005282 vitamin D3 Nutrition 0.000 description 1
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- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 1
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- 229940021056 vitamin d3 Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 239000010497 wheat germ oil Substances 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
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- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- CPYIZQLXMGRKSW-UHFFFAOYSA-N zinc;iron(3+);oxygen(2-) Chemical compound [O-2].[O-2].[O-2].[O-2].[Fe+3].[Fe+3].[Zn+2] CPYIZQLXMGRKSW-UHFFFAOYSA-N 0.000 description 1
- 229910001928 zirconium oxide Inorganic materials 0.000 description 1
- 239000004711 α-olefin Substances 0.000 description 1
- 229930007845 β-thujaplicin Natural products 0.000 description 1
Images
Definitions
- the purpose of the present invention is to provide the innate immunomodulatory effect of ginseng fruit polysaccharide in immunosuppressed conditions.
- the immune system is an organization of cells and molecules that defends the body against infection and consists of two major subsystems: the innate immune system and the adaptive immune system. Changes in immune responses and immune cell populations such as immune organ markers (bone marrow, spleen, thymus) and immune cell populations (macrophages, spleen cells, neutrophils, natural killer cells) are essential mechanisms to improve the body's immune function.
- This immune response produces a variety of cytokines (e.g., interferon- ⁇ (IFN- ⁇ ), tumor necrosis factor- ⁇ (TNF- ⁇ ), interleukin-2 (IL-2), and IL-6), which have various immunomodulatory functions. Activates other inflammatory mediators, and natural products of plant extracts have the ability to stimulate the host's immune system and could be a potential treatment alternative to chemotherapy.
- IFN- ⁇ interferon- ⁇
- TNF- ⁇ tumor necrosis factor- ⁇
- IL-2 interleukin-2
- IL-6 interleukin-6
- An immune response is a series of reactions that activated immune cells produce against foreign and endogenous substances (antigens).
- host cells secrete factors such as cytokines that cause the inflammatory response to overcome the infection, thereby causing an inflammatory response.
- cytokines that cause the inflammatory response to overcome the infection, thereby causing an inflammatory response. It is known that when these microorganisms and foreign substances in living organisms are recognized by immune cells, the immune cells are activated, and the activated immune cells also secrete many factors that cause an inflammatory response, causing an inflammatory response.
- innate immunity which a person has from birth
- acquired immunity which is acquired through adaptation to life.
- innate immunity is also called 'natural immunity', and reacts non-specifically to antigens and does not show any special memory function.
- the innate immune system includes skin, mucus tissue, and acidic stomach acid that block the invasion of antigens, and phagocytes such as macrophages and polymorphonuclear leukocytes that are responsible for phagocytosis to remove invaders that have entered the body.
- phagocytes such as macrophages and polymorphonuclear leukocytes that are responsible for phagocytosis to remove invaders that have entered the body.
- This innate immunity actually protects against most infections.
- acquired immunity has a memory function for the first invading antigen, and when invasion occurs again, it has the characteristic of reacting specifically and effectively eliminating the antigen.
- Anticancer drugs such as cyclophosphamide (CY) have been used in chemotherapy to treat cancer, autoimmune diseases, and immune-mediated diseases. However, it can cause serious side effects such as cytotoxicity and oxidative stress, and it also damages the DNA of normal cells, causing significant immunosuppression. In these cases, it is important to restore the immunosuppressed state, and the cyclophosphamide (CY)-induced immunosuppressed mouse model has been used in many previous studies to evaluate the immunomodulatory effects of functional substances.
- CY cyclophosphamide
- Polysaccharides are a type of bioactive polymer material with beneficial pharmacological activities, including anti-tumor, antioxidant, anti-diabetic and immunomodulatory effects.
- Korean ginseng (Panax ginseng C. A. Meyer) has been reported to have immune-modulating, anti-cancer, anti-aging, and anti-diabetic effects in Korea, China, and several East Asian countries.
- Polysaccharides from ginseng can exert immunomodulatory activities in both in vitro and in vivo systems, and crude polysaccharides from American ginseng ( P. quinquefolius ) not only promote spleen lymphocyte proliferation but also promote macrophage phagocytosis and nitric oxide. NO) production can be increased.
- the present inventors found that after cellulase- and ⁇ -amylase-based extraction, the polysaccharides of Korean ginseng induced increases in spleen and thymus indices, lymphocyte proliferation, white blood cell count, and NK cell activity in cyclophosphamide (CY)-induced immunosuppressed mice, thereby improving immunity. By confirming that there was an enhancement effect, the present invention was completed.
- the purpose of the present invention is to provide a food composition for enhancing immunity containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- an object of the present invention is to provide a cosmetic composition for enhancing immunity containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- the purpose of the present invention is to provide a feed composition for immunity enhancement containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- the present invention provides a food composition for enhancing immunity containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- the present invention provides a cosmetic composition for enhancing immunity containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- the present invention provides a feed composition for immunity enhancement containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- the Korean ginseng fruit polysaccharide of the present invention was confirmed to have an immunity-increasing effect. Specifically, the Korean ginseng fruit polysaccharide of the present invention was administered to immunosuppressed mice induced under anti-cancer, immunosuppression, or infection conditions to increase spleen size and spleen indices, and it was confirmed that NK cell activity was activated. In addition, it was confirmed that lymphocytes in the spleen were proliferated, the expression of immune-related genes was increased, and both innate and acquired immunity were confirmed to be enhanced, so it can be usefully used in related projects.
- FIG 1 is a diagram showing the spleen size of mice treated with various concentrations of Korean ginseng fruit polysaccharide (GBP).
- GBP Korean ginseng fruit polysaccharide
- FIG 2 is a diagram showing the spleen index of mice treated with various concentrations of Korean ginseng fruit polysaccharide (GBP).
- GBP Korean ginseng fruit polysaccharide
- FIG 3 is a diagram showing spleen cell proliferation in mice treated with Korean ginseng fruit polysaccharide (GBP) and induced with concanavalin A (Con A) or lipopolysaccharide (LPS).
- GBP Korean ginseng fruit polysaccharide
- Con A concanavalin A
- LPS lipopolysaccharide
- FIG 4 is a diagram showing NK cell activity by treatment with Korean ginseng fruit polysaccharide (GBP).
- FIG. 5 is a diagram showing mRMA expression of IL-1 ⁇ by treatment with Korean ginseng fruit polysaccharide (GBP).
- FIG. 6 is a diagram showing mRMA expression of IL-2 by treatment with Korean ginseng fruit polysaccharide (GBP).
- FIG. 7 is a diagram showing mRMA expression of IL-4 by treatment with Korean ginseng fruit polysaccharide (GBP).
- FIG 8 is a diagram showing mRMA expression of IL-6 by treatment with Korean ginseng fruit polysaccharide (GBP).
- FIG 9 is a diagram showing mRMA expression of TNF- ⁇ by treatment with Korean ginseng fruit polysaccharide (GBP).
- FIG 10 is a diagram showing mRMA expression of IFN- ⁇ by treatment with Korean ginseng fruit polysaccharide (GBP).
- FIG 11 is a diagram showing mRMA expression of TLR-4 by treatment with Korean ginseng fruit polysaccharide (GBP).
- Figure 12 is a diagram showing mRMA expression of COX-2 by treatment with Korean ginseng fruit polysaccharide (GBP).
- the present invention provides a food composition for enhancing immunity containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- the Korean ginseng fruit extract is extracted with any solvent selected from the group consisting of water, C1 to C4 lower alcohol, or a mixed solvent thereof, but is not limited thereto.
- the Korean ginseng fruit polysaccharide consists of rhamnose, arabinose, mannose, glucose, and galactose, but is not limited thereto.
- the Korean ginseng fruit polysaccharide has a molecular weight of 328.4 and 54.2 ⁇ 10 3 (g/mol), but is not limited thereto.
- the Korean ginseng fruit polysaccharide contains 4.0% of rhamnose, 19.8% of arabinose, 2.2% of mannose, 27.7% of glucose, and 46.3% of galactose, but is not limited thereto.
- the composition increases spleen size and spleen parameters, but is not limited thereto.
- the composition increases T lymphocytes and B lymphocytes, but is not limited thereto.
- the composition increases NK cell activity, but is not limited thereto.
- the composition increases immune-related genes, but is not limited thereto.
- the immune-related gene is selected from the group consisting of IL-2, IL-4, IL-6, TNF- ⁇ , IFN- ⁇ , TLR-4, and COX-2, but is not limited thereto. .
- the immunity is reduced immunity due to anticancer drugs, immunosuppressants, or infection, but is not limited thereto.
- the food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients like a typical food composition.
- Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
- the above-described flavoring agents include natural flavoring agents (thaumatin), stevia extracts (e.g. rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).
- the food composition of the present invention can be formulated in the same way as the pharmaceutical composition and used as a functional food or added to various foods.
- Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes, health supplements, etc. There is.
- the food composition contains, in addition to the extract as an active ingredient, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, It may contain alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.
- the food composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages.
- the functional food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc.
- 'health functional food composition' refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with Act No. 6727 on Health Functional Food, and refers to food that is related to the structure and function of the human body. It means taking it for the purpose of controlling nutrients or obtaining useful health effects such as physiological effects.
- the health functional food of the present invention may contain common food additives, and its suitability as a food additive is determined in accordance with the general provisions and general test methods of the food additive code approved by the Food and Drug Administration, unless otherwise specified. Judgment is made according to specifications and standards.
- Items listed in the 'Food Additive Code' include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as dark pigment, licorice extract, crystalline cellulose, kaoliang pigment, and guar gum; Examples include mixed preparations such as sodium L-glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations.
- the health functional food in tablet form is made by granulating a mixture of the active ingredient of the present invention with excipients, binders, disintegrants and other additives in a conventional manner, and then adding a lubricant and compression molding, or The mixture can be directly compression molded.
- the health functional food in the form of tablets may contain flavoring agents, etc., if necessary.
- hard capsules can be manufactured by filling a regular hard capsule with a mixture of the active ingredient of the present invention with additives such as excipients
- soft capsules can be prepared by mixing the active ingredient of the present invention with additives such as excipients. It can be manufactured by filling the mixture with a capsule base such as gelatin.
- the soft capsule may contain plasticizers such as glycerin or sorbitol, colorants, preservatives, etc., if necessary.
- the health functional food in the form of a pill can be prepared by molding a mixture of the active ingredient of the present invention and excipients, binders, disintegrants, etc., using a known method. If necessary, it can be coated with white sugar or other coating agent. Alternatively, the surface can be coated with substances such as starch or talc.
- Health functional food in the form of granules can be manufactured into granules by mixing a mixture of excipients, binders, disintegrants, etc. of the active ingredients of the present invention by a known method, and may contain flavoring agents, flavoring agents, etc., if necessary. You can.
- %' used to indicate the concentration of a specific substance means (w/w) % for solid/solid, (w/v) % for solid/liquid, and Liquid/liquid is (v/v) %.
- the present invention provides a cosmetic composition for enhancing immunity containing Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- Products to which this composition can be added include, for example, cosmetics such as various creams, lotions, skins, and essences, as well as shampoos, rinses, cleansers, face washes, soaps, treatments, packs, and beauty essences.
- the cosmetic of the present invention includes a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids, and seaweed extracts.
- Water-soluble vitamins may be any that can be mixed into cosmetics, but preferably include vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, and vitamin H. and their salts (thiamine hydrochloride, sodium ascorbate, etc.) or derivatives (sodium ascorbic acid-2-phosphate, magnesium ascorbic acid-2-phosphate, etc.) are also water-soluble vitamins that can be used in the present invention. Included. Water-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification from microbial cultures, enzymatic methods, or chemical synthesis.
- Oil-soluble vitamins may be any that can be mixed into cosmetics, but preferably include vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol), etc. , their derivatives (ascorbine palmitate, ascorbine stearate, ascorbine dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol nicotinate, vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl ethyl ether, etc.) are also included in the oil-soluble vitamins used in the present invention. Oil-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification from microbial cultures, enzymatic or chemical synthesis.
- the polymer peptide may be any peptide that can be blended into cosmetics, but preferably includes collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, and keratin.
- High molecular weight peptides can be purified and obtained by conventional methods such as purification from microbial culture media, enzymatic methods, or chemical synthesis, or they can usually be purified and used from natural products such as dermis of pigs or cows or silk fiber of silkworms.
- the high molecular polysaccharide may be any one that can be blended into cosmetics, but preferably includes hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or its salts (sodium salt, etc.).
- chondroitin sulfate or its salt can usually be purified and used from mammals or fish.
- the sphingolipid may be any one that can be blended into cosmetics, but preferred examples include ceramide, phytosphingosine, and sphingolipid. Sphingolipids can usually be purified by conventional methods from mammals, fish, shellfish, yeast, or plants, or obtained by chemical synthesis.
- the cosmetic of the present invention may contain other ingredients usually blended in cosmetics as needed.
- Other ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohol, colorants, fragrances, Examples include blood circulation promoters, cooling agents, antiperspirants, and purified water.
- oil and fat components include ester-based fats and oils, hydrocarbon-based fats and oils, silicone-based fats and oils, fluorine-based fats and oils, animal fats and oils, and vegetable fats and oils.
- Ester-based oils include glyceryl tri2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid.
- Reyl isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, cetoester 2-ethylhexanoate.
- triisocetyl citrate triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, malic acid.
- Diisostearyl acid 2-ethylhexyl hydroxystearate, di2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate.
- cholesteryl hydroxystearate cholesteryl oleate, dihydrocholesteryl oleate, phytsteryl isostearate, phytsteryl oleate, 12-stealoyl hydroxystearate isocetyl, 12-stealoyl Ester systems such as stearyl monohydroxystearate and isostearyl 12-stealoylhydroxystearate can be mentioned.
- Hydrocarbon-based fats and oils include squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybudene, microcrystalline wax, and petroleum jelly.
- Silicone oils include polymethyl silicone, methylphenyl silicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane and methylcetyloxysiloxane copolymer, dimethylsiloxane and methylstealoxysiloxane copolymer, and alkyl. Modified silicone oil, amino-modified silicone oil, etc. can be mentioned.
- fluorine-based fats and oils examples include perfluoropolyether.
- Animal or plant oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, birdflower oil, soybean oil, corn oil, rapeseed oil, apricot oil, palm kernel oil, palm oil, castor oil, sunflower oil, and grape seed oil. , cottonseed oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, shea butter, walnut colostrum oil, marker damia nut oil, meadow oil, egg yolk oil, beef tallow, horse oil, mink oil, orange rape oil, jojoba oil. , animal or plant oils such as candelier wax, carnaba wax, liquid lanolin, and hydrogenated castor oil.
- Moisturizers include water-soluble low-molecular-weight moisturizers, oil-soluble molecular moisturizers, water-soluble polymers, and fat-soluble polymers.
- fat-soluble low-molecular-weight moisturizers examples include cholesterol and cholesterol esters.
- Water-soluble polymers include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water-soluble chitin, chitosan, and dextrin. You can.
- oil-soluble polymers examples include polyvinylpyrrolidone and eicosene copolymers, polyvinylpyrrolidone and hexadecene copolymers, nitrocellulose, dextrin fatty acid ester, and polymer silicone.
- emollients include long-chain acyl glutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, and lanolin fatty acid cholesteryl ester.
- surfactants include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants.
- Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE.
- Anionic surfactants include fatty acid soap, alpha-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkyl amide sulfate, alkyl phosphate, POE alkyl phosphate industry, alkyl amide.
- alkyloyl alkyl taurine salt N-acylamino acid salt
- POE alkyl ether carboxylate alkyl sulfosuccinate, sodium alkyl sulfoacetate, acylated hydrolyzed collagen peptide salt, perfluoroalkyl phosphate ester, etc. there is.
- Cationic surfactants include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, and chloride.
- Examples include benzalkonium, diethylaminoethylamide stearate, dimethylaminopropyl stearate, and quaternary ammonium salts of lanolin derivatives.
- Amphoteric surfactants include carboxy beta type, amide beta type, sulfo beta type, hydroxy sulfo beta type, amide sulfo betaine type, phosphobetain type, aminocarboxylate type, imidazoline derivative type, and amidamine type. Amphoteric surfactants, etc. can be mentioned.
- Organic and inorganic pigments include silicic acid, anhydrous silicic acid, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, and aluminum oxide.
- Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene and styrene copolymer, Examples include organic pigments such as silk powder, cellulose, CI pigment yellow, and CI pigment orange, and complex pigments of these inorganic pigments and organic pigments.
- organic powder examples include metal soap such as calcium stearate; Alkyl phosphate metal salts such as sodium zinc cetilate, zinc laurylate, and calcium laurylate; Acyl amino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc, and N-lauroyl glycine calcium; Amidesulfonic acid polyvalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylizine, N-alpha-paritoylolnithine, N-alpha-lauroylarginine, N-alpha-hydrogenated beef tallow fatty acid acylarginine, etc.
- metal soap such as calcium stearate
- Alkyl phosphate metal salts such as
- N-acyl polypeptides such as N-lauroylglycylglycine
- Alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid
- Examples include polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene and styrene copolymers, and ethylene tetrafluoride.
- UV absorbers include para-aminobenzoic acid, ethyl para-aminobenzoate, amyl para-aminobenzoate, octyl para-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, and benzyl cinnamate.
- Disinfectants include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zincphyllithione, benzalkonium chloride, and photosensitive.
- Sub-No. 301, mononitroguaiacol sodium, undecirenic acid, etc. can be mentioned.
- Antioxidants include butylhydroxyanisole, propyl gallate, and elisorbic acid.
- pH adjusters examples include citric acid, sodium citrate, malic acid, sodium malate, fumal acid, sodium fumalate, succinic acid, sodium succinate, sodium hydroxide, and sodium monohydrogen phosphate.
- alcohol examples include higher alcohols such as cetyl alcohol.
- ingredients that may be added are not limited to this, and any of the above ingredients can be blended within the range that does not impair the purpose and effect of the present invention.
- the cosmetic of the present invention may take the form of a solution, emulsion, viscous mixture, etc.
- the ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions as active ingredients, for example, conventional auxiliaries and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances. Includes.
- the cosmetic composition for skin whitening and skin aging prevention of the present invention can be manufactured in any formulation commonly manufactured in the art, for example, emulsion, cream, lotion, pack, foundation, lotion, beauty essence, hair cosmetics, etc. You can.
- the cosmetic composition of the present invention includes skin lotion, skin softener, skin toner, milk lotion, astringent, lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, foundation, essence, nutritional essence, It includes formulations of packs, soaps, cleansing foams, cleansing lotions, cleansing creams, hair lotions, hair tonics, hair essences, hair shampoos, hair rinses, hair treatments, body lotions, and body cleansers.
- the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier ingredient. You can.
- the formulation of the present invention is a powder or spray
- lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient.
- chlorofluorohydrocarbon and propane may be used as carrier ingredients.
- May contain propellants such as butane or dimethyl ether.
- a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, fatty esters of glycerol, fatty acid esters of polyethylene glycol or sorbitan.
- the carrier ingredients include water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, and microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
- a liquid diluent such as ethanol or propylene glycol
- a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester
- microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
- the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, and fatty acid amide.
- Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative, or ethoxylated glycerol fatty acid ester can be used.
- the present invention provides a feed composition for immunity enhancement comprising Korean ginseng berry polysaccharides (GBP) as an active ingredient.
- GBP Korean ginseng berry polysaccharides
- the feed composition of the present invention replaces existing antibiotics and inhibits the growth of harmful food pathogens, which is expected to improve the health of animals, improve weight gain and meat quality of livestock, and increase milk production and immunity. You can.
- the feed composition of the present invention can be manufactured in the form of fermented feed, compounded feed, pellet form, silage, etc.
- the fermented feed can be manufactured by fermenting organic matter by adding various microorganisms or enzymes other than the peptide of the present invention, and the compounded feed can be manufactured by mixing various types of general feed with the peptide of the present invention.
- Pellet-type feed can be manufactured by applying heat and pressure to the above-mentioned mixed feed, etc.
- silage in a pellet machine, and silage can be manufactured by fermenting the green feed with the microorganism according to the present invention.
- Wet fermented feed collects and transports organic matter such as food waste, mixes excipients for sterilization and moisture control in a certain ratio, and then ferments for more than 24 hours at a temperature suitable for fermentation to ensure that the moisture content is about 70%. It can be manufactured by adjusting it.
- Fermented dried feed can be manufactured by adjusting wet fermented feed to an additional drying process so that the moisture content is about 30% to 40%.
- the feed composition of the present invention may further include ingredients added to conventional feed.
- ingredients added to such feed may include grain powder, meat powder, and beans.
- the grain powder may be one or more selected from rice flour, wheat flour, barley flour, and corn flour.
- the meat powder may be a meat powder obtained by pulverizing any one or more selected from chicken, beef, pork, and ostrich meat.
- the pulses may be one or more types selected from soybeans, kidney beans, peas, and black beans.
- the feed composition of the present invention can add one or more selected from nutrients and minerals to increase the nutritional value of the feed, and feed quality.
- it may contain one or more selected from anti-fungal agents, antioxidants, anti-coagulants, emulsifiers, and binders.
- mice Six-week-old male BALB/c mice (21-23 g) were purchased from the Central Research Institute, Animal Inc. (Korea), the temperature of the mice was maintained at 22 ⁇ 2 o C, and a 12-hour/12-hour dark/light cycle was used as standard. It was accepted with conditions.
- mice were provided with a standard laboratory diet and water for at least 1 week before the experiment. All experimental procedures were approved by the Gangneung-Wonju National University committee.
- Korean ginseng fruit powder (20 g) purchased from Boeun, Chungcheong-do, Korea was extracted with 99% ethanol for 3 hours, the extract was centrifuged at 4000 xg for 15 minutes, and the precipitate was dried at room temperature. Distilled water was added to the dried precipitate and extracted at 100°C for 3 hours.
- the polysaccharide extract was obtained by adding 99% ethanol, centrifugation, filtration, washing with acetone, and lyophilization on the dried polysaccharide precipitate to produce crude polysaccharide (GBP-C).
- GBP-C was fractionated using ion exchange chromatography on a DEAE Sepharose fast flow column (17-0709-01; GE Healthcare Bio-Science AB, Uppsala, Sweden), containing rhamnose (4.0%), arabinose ( Considered as a domestically produced product with a molecular weight of 328.4 and 54.2 Korean ginseng berry polysaccharides (GBP) were prepared.
- Cyclophosphamide was used to induce an immunosuppression model by intraperitoneal (IP) injection at a dose of 80 mg/kg body weight (BW), and levamisole (LEV) was used to induce a cell-mediated, That is, as an immunomodulator capable of regulating T cell-induced immunity, a dose of 40 mg/kg BW was used as a positive control, and commercial red ginseng syrup was used as a positive control at a dose of 100 mg/kg BW. In addition, commercially available ginseng products were used, and for the samples in Table 1, mice were sacrificed 24 hours after the last treatment.
- the spleen was dissected and weighed.
- the spleen index was calculated according to the following formula.
- Spleen index (mg/g) spleen weight/mouse body weight
- splenocytes were weighed and the spleen of the mouse was placed in ice-cold PBS for spleen cell isolation.
- Mouse splenocytes were extracted using RBC Lysis Buffer (eBioscience, USA). According to the manufacturer's instructions, spleen cells were cultured in RPMI1640 medium (Gibco Laboratories, USA) supplemented with 1% fetal bovine serum, 1% streptomycin (100 ⁇ g/ml), and penicillin (100 IU/ml) (Welgene, Korea). The concentration was adjusted to 2 ⁇ 10 6 cells/ml.
- EZ-Cytox Cell Viability Assay Kit (Daeil Lab Service, Korea) was used to measure splenic lymphocyte proliferation.
- Spleen cells were seeded in a 96-well plate and cultured at 37°C, 5% CO 2 for 1 hour. Cultured spleen cells were stimulated with 5 ⁇ g/ml concanavalin A (Con A) as a T cell mitogen or 10 ⁇ g/ml lipopolysaccharide (LPS) as a B cell mitogen. After incubation at 37°C with 5% CO 2 for 48 hours, 25 ⁇ l EZ-Cytox reagent was added to each well and the plate was incubated for an additional 1 hour at 37°C with 5% CO 2 . Absorbance was measured at 450 nm using a microplate reader (EL800; BioTek, USA).
- Con A concanavalin A
- LPS lipopolysaccharide
- Spleen cells were cultured with YAC-1 cells (Korean Cell Line Bank) or co-cultured without YAC-1 cells to obtain a ratio of effector cells to NK-sensitive target cells of 50:1. After 24 hours of incubation, the supernatant was collected after centrifugation. The activity of NK cells was evaluated using the CytoTox 96 Non-Radioactive Cytotoxicity Assay kit (Promega, USA) at 250 ⁇ g for 5 minutes. Then, the absorbance at 490 nm was measured.
- Tri reagent (Molecular Research Center, Inc., USA) was used to extract total RNA from splenic lymphocytes. Total RNA was reverse transcribed into cDNA using a high-capacity cDNA reverse transcription kit (Applied Biosystems, USA). To amplify cDNA, cDNA was subjected to qRT-PCR using TB Green Premix Ex Taq II (Takara Bio Inc., Japan) in QuantStudio 3 Flex Real-Time PCR System (Thermo Fisher Scientific, USA). Amplification conditions were as follows: initial denaturation at 95°C for 30 s, followed by 40 cycles of 95°C for 5 s and 60°C for 34 s. Primer sequences are listed in Table 2.
- cyclophosphamide (CY) 80 mg/kg was administered to immunosuppressed mice and Levamisole (Levamisole) was administered to immunosuppressed mice.
- LEV 40 mg/kg treatment group, red ginseng (100 mg/kg) treatment group, and Korean ginseng fruit polysaccharide (GBP) treatment group at different concentrations (50, 100, 250, and 500 mg/kg BW). and spleen indices were confirmed.
- the cyclophosphamide (CY) treated group showed a decrease in both spleen size and indicators, while the levamisole (LEV) treated group and the red ginseng treated group had increased spleen size and indicators.
- spleen size and indices were confirmed to be increased in a concentration-dependent manner (50, 100, 250, and 500 mg/kg BW).
- the Korean ginseng fruit polysaccharide (GBP) of the present invention increases immunity by increasing spleen size ( Figure 1) and spleen index ( Figure 2).
- GBP Korean ginseng fruit polysaccharide
- Concanavalin A Con A
- lipopolysaccharide LPS
- CY cyclophosphamide
- the Korean ginseng fruit polysaccharide (GBP) of the present invention increases immunity by increasing the proliferation of T lymphocytes and B lymphocytes.
- NK cell activity at concentrations of 100, 250, and 500 mg/kg BW of Korean ginseng fruit polysaccharide (GBP) was confirmed to be significantly higher than that of the cyclophosphamide (CY) treated group in a concentration-dependent manner.
- the Korean ginseng fruit polysaccharide (GBP) of the present invention increases immunity by increasing NK cell activity.
- Example 4 Effect of Korean ginseng fruit polysaccharide (GBP) on immune-related gene expression.
- GBP Korean ginseng fruit polysaccharide
- Korean ginseng fruit polysaccharide (GBP) was used as a T cell mitogen, the mitogen concanavalin A (Con A), or a B cell mitogen.
- Con A the mitogen concanavalin A
- B cell mitogen the mitogen concanavalin A
- LPS lipopolysaccharide
- Korean ginseng fruit polysaccharide (250 and 500 mg/kg BW) increased the expression level of immune-related genes compared to the normal group.
- the Korean ginseng fruit polysaccharide (GBP) of the present invention increases immunity by increasing immune-related genes.
- Korean ginseng fruit polysaccharide (GBP), cyclophosphamide (CY), exhibits potent immunomodulatory properties in induced immunosuppressed mice, and treatment with Korean ginseng fruit polysaccharide (GBP) in immunosuppressed mice improves spleen indices and improves the immune response. has been strengthened.
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Abstract
본 발명의 목적은 고려인삼열매의 다당류을 유효성분으로 포함하는 면역강화 조성물을 제공하는 것으로서, 본 발명의 고려 인삼 열매 다당류는 면역 증가 효과를 확인 하였다. 구체적으로 항암, 면역억제 또는 감염 조건에서 유발된 면역억제 마우스에 본 발명의 고려 인삼 열매 다당류를 투여하여 비장 크기 및 비장 지표를 증가 시켰으며, NK세포 활동을 활성화 시키는 것을 확인하였다. 또한, 비장의 림프구를 증식 시켰으며, 면역관련 유전자의 발현을 증가시키는 것을 확인하였고, 선천면역 또는 후천면역을 모두 증강시킴을 확인하여, 이와 관련된 사업에 유용하게 사용될 수 있다.
Description
본 발명의 목적은 면역저하상태에서의 인삼 열매 다당류의 선천성 면역조절 효과를 제공하는 것이다.
면역 체계는 감염으로부터 신체를 방어하는 세포와 분자의 조직으로서,선천 면역 체계와 적응 면역 체계의 두 가지 주요 하위 시스템으로 구성된다. 면역 기관 지표(골수, 비장, 흉선)와 같은 면역 반응과 면역 세포 집단(대식세포, 비장 세포, 호중구, 자연 살해 세포)의 변화는 신체의 면역 기능을 향상시키는 데 필수적인 메커니즘이다. 이러한 면역 반응은 다양한 사이토카인(예: 인터페론-γ(IFN-γ), 종양 괴사 인자-α(TNF-α), 인터루킨-2(IL-2) 및 IL-6)을 생성하여 다양한 면역 조절 기능을 활성화하고 다른 염증 매개체이며, 식물 추출물의 천연 제품은 숙주의 면역 체계를 자극하는 능력이 있으며 화학 요법을 대체할 수 있는 잠재적인 치료법이 될 수 있다.
면역반응은 활성화된 면역세포가 외래성 및 내인성 물질(항원)에 대하여 일으키는 일련의 반응이다. 세균, 바이러스 등을 포함한 미생물 및 생체의 이물질 등이 생체로 유입되면, 숙주세포는 감염을 극복하기 위해 염증반응의 원인이 되는 사이토카인 등의 인자를 분비하여 염증반응을 유발시킨다. 그리고 이러한 미생물 및 생체의 이물질 등이 면역세포에 의해 인식이 되면 면역세포가 활성화되고, 활성화된 면역세포에서도 염증반응의 원인이되는 많은 인자를 분비하여 염증반응을 유발한다고 알려져 있다.
이러한 면역반응은 태어날 때부터 가지고 있는 선천면역 (innate immunity)과 후천적으로 생활 등에 적응되어 얻어지는 후천면역(acquired immunity)로 구분된다. 여기서, 선천면역은 일명 '자연면역'이라고도 하는 것으로, 항원에 대하여 비특이적으로 반응하며 특별한 기억작용은 보이지 않는다. 선천면역체계로는 항원의 침입을 차단하는 피부, 점액조직, 상산성의 위산, 체내로 들어온 침입자들을 제거하는 식균작용을 담당하는 대식세포(macrophage), 다형핵 백혈구 등과 같은 식세포 등이 있다. 이러한 선천면역은 실제 대부분의 감염을 방어한다. 한편, 후천면역은 처음 침입한 항원에 대한 기억작용이 있어, 다시 침입이 발생하였을 때, 특이적으로 반응하여 효과적으로 항원을 제거하는 특징이 있다.
이러한 면역기능이 저하되면, 천식, 계절성 또는 통년성 비염, 알러지성 비염, 결막염, 아토피성 피부염, 두드러기, 적혈구의 용혈, 급성 사구체 신염, 감기, 만성 피로 및 암 등 다양한 면역질환이 발생할 수 있다. 이러한면역질환은 치료보다 예방이 중요한데, 현재 시판되고 있는 치료제는 예방을 위하여 복용하기에는 어려운 것이 현실이다. 이에 부작용의 가능성이 있는 면역증강제의 복용 없이, 면역기능 장애를 예방 및 치료할 수 있는 방법이 요구되었다.
시클로포스파미드(cyclophosphamide, CY)와 같은 항암제는 암, 자가면역 질환 및 면역 매개 질환을 치료하기 위한 화학요법에 사용되어 왔다. 그러나, 세포 독성 및 산화 스트레스와 같은 심각한 부작용을 야기시킬수 있으며, 또한, 정상 세포의 DNA를 손상시켜 상당한 면역 억제 상태를 야기 시킨다. 이러한 경우에는 면역 억제 상태를 회복시키는 것이 중요하며, 시클로포스파미드(CY) 유도 면역 억제 마우스 모델은 기능성 물질의 면역 조절 효과를 평가하기 위해 많은 이전 연구에서 사용되왔다.
다당류(Polysaccharides)는 유익한 약리 활성을 가진 생체 활성 고분자 물질의 한 종류로서, 항 종양, 항산화, 항 당뇨병 및 면역 조절 효과를 가지고 있다.
고려인삼(Panax ginseng C. A. Meyer)은 한국, 중국 및 몇몇 동아시아 국가들에 인삼 열매는 면역 조절을 나타내는 것으로 보고되어 왔고, 항암, 항노화, 항당뇨 효과가 있다고 보고 되었다.
인삼의 다당류는 in vitro 및 in vivo 시스템 모두에서 면역 조절 활성을 발휘할 수 있으며, 서양삼(P. quinquefolius)의 조다당류는 비장 림프구 증식뿐만 아니라 대식세포 포식작용(pacrophage phagocytosis) 및 산화질소(nitric oxide, NO) 생산을 증가시킬 수 있다.
이에 본 발명자는 셀룰라아제 및 α-아밀라아제 기반 추출 후 고려인삼의 다당류는 시클로포스파미드(CY) 유발 면역억제된 마우스의 비장 및 흉선 지수, 림프구 증식, 백혈구 수, NK 세포 활성 증가를 유도하여, 면역 증진 효과가 있음을 확인 하여, 본 발명을 완성 하였다.
본 발명의 목적은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 식품 조성물을 제공하는 것이다.
또한, 본 발명의 목적은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 화장료 조성물을 제공하는 것이다.
아울러, 본 발명의 목적은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 사료 조성물을 제공하는 것이다.
본 발명은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 식품 조성물을 제공한다.
또한, 본 발명은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 화장료 조성물을 제공한다.
아울러, 본 발명은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 사료 조성물을 제공한다.
본 발명의 고려 인삼 열매 다당류는 면역 증가 효과를 확인 하였다. 구체적으로 항암, 면역억제 또는 감염 조건에서 유발된 면역억제 마우스에 본 발명의 고려 인삼 열매 다당류를 투여하여 비장 크기 및 비장 지표를 증가 시켰으며, NK세포 활동을 활성화 시키는 것을 확인하였다. 또한, 비장의 림프구를 증식 시켰으며, 면역관련 유전자의 발현을 증가시키는 것을 확인하였고, 선천면역 또는 후천면역을 모두 증강시킴을 확인하여, 이와 관련된 사업에 유용하게 사용될 수 있다.
도 1은 고려인삼열매 다당류(GBP)가 다양한 농도로 처리된 마우스의 비장 크기를 나타낸 도이다.
도 2는 고려인삼열매 다당류(GBP)가 다양한 농도로 처리된 마우스의 비장 지수를 나타낸 도이다.
도 3은 고려인삼열매 다당류(GBP)가 처리되고 콘카나발린 A(Con A) 또는 리포폴리사카라이드(LPS)로 유발된 마우스의 비장 세포 증식을 나타낸 도이다.
도 4는 고려인삼열매 다당류(GBP)의 처리에 의한 NK 세포 활성을 나타낸 도이다.
도 5는 고려인삼열매 다당류(GBP)의 처리에 의한 IL-1β의 mRMA 발현을 나타낸 도이다.
도 6은 고려인삼열매 다당류(GBP)의 처리에 의한 IL-2의 mRMA 발현을 나타낸 도이다.
도 7은 고려인삼열매 다당류(GBP)의 처리에 의한 IL-4의 mRMA 발현을 나타낸 도이다.
도 8은 고려인삼열매 다당류(GBP)의 처리에 의한 IL-6의 mRMA 발현을 나타낸 도이다.
도 9는 고려인삼열매 다당류(GBP)의 처리에 의한 TNF-α의 mRMA 발현을 나타낸 도이다.
도 10은 고려인삼열매 다당류(GBP)의 처리에 의한 IFN-γ의 mRMA 발현을 나타낸 도이다.
도 11은 고려인삼열매 다당류(GBP)의 처리에 의한 TLR-4의 mRMA 발현을 나타낸 도이다.
도 12는 고려인삼열매 다당류(GBP)의 처리에 의한 COX-2의 mRMA 발현을 나타낸 도이다.
이하, 첨부된 도면을 참조하여 본 발명의 구현예로 본 발명을 상세히 설명하기로 한다. 다만, 하기 구현의 예는 본 발명에 대한 예시로 제시되는 것으로, 당업자에게 주지 저명한 기술 또는 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우에는 그 상세한 설명을 생략할 수 있고, 이에 의해 본 발명이 제한되지는 않는다. 본 발명은 후술하는 특허청구범위의 기재 및 그로부터 해석되는 균등 범주 내에서 다양한 변형 및 응용이 가능하다.
또한, 본 명세서에서 사용되는 용어(terminology)들은 본 발명의 바람직한 실시예를 적절히 표현하기 위해 사용된 용어들로서, 이는 사용자, 운용자의 의도 또는 본 발명이 속하는 분야의 관례 등에 따라 달라질 수 있다. 따라서 본 용어들에 대한 정의는 본 명세서 전반에 걸친 내용을 토대로 내려져야 할 것이다. 명세서 전체에서, 어떤 부분이 어떤 구성요소를 “포함”한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성요소를 제외하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다.
본 발명은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 식품 조성물을 제공한다.
일 실시예에 있어서, 상기 고려 인삼 열매 추출물은 물, C1 내지 C4의 저급 알코올 또는 이들의 혼합용매로 이루어 진 군에서 선택된 어느 하나의 용매로 추출된 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 고려 인삼 열매 다당류는 람노스(rhamnose), 아라비노스(arabinose), 만노스(mannose), 글루코스(glucose) 및 갈락토스(galactose)로 이루어진 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 고려 인삼 열매 다당류의 분자량 328.4 및 54.2 × 103(g/mol)로 구성하는 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 고려 인삼 열매 다당류는 람노스의 4.0%, 아라비노스는 19.8%, 만노스는 2.2%, 글루코스 27.7% 및 갈락토스는 46.3%로 포함된 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 조성물은 비장의 크기 및 비장 지표를 증가시키는 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 조성물은 T 림프구 및 B 림프구를 증가시키는 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 조성물은 NK 세포 활성을 증가시키는 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 조성물은 면역 관련 유전자를 증가 시키는 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 면역 관련 유전자는 IL-2, IL-4, IL-6, TNF-α, IFN-γ, TLR-4 및 COX-2로 이루어진 군에서 선택되는 것이나, 이에 제한되지 않는다.
일 실시예에 있어서, 상기 면역은 항암제, 면역억제제 또는 감염에 의한 면역이 저하된 것이나, 이에 제한되지 않는다.
본 발명의 식품 조성물은 유효성분인 추출물을 함유하는 것 외에 통상의 식품 조성물과 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다.
상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 향미제는 천연 향미제 (타우마틴), 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제 (사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 본 발명의 식품 조성물은 상기 약학적 조성물과 동일한 방식으로 제제화되어 기능성 식품으로 이용하거나, 각종 식품에 첨가할 수 있다. 본 발명의 조성물을 첨가할 수 있는 식품으로는 예를 들어, 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류 등이 있다.
또한 상기 식품 조성물은 유효성분인 추출물 외에 여러 가지 영양제, 비타민, 광물 (전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제 (치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 식품 조성물은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다.
본 발명의 기능성 식품 조성물은, 정제, 캅셀, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공될 수 있다. 본 발명에서 '건강기능성 식품 조성물'이라 함은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 말하며, 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다. 본 발명의 건강기능식품은 통상의 식품 첨가물을 포함할 수 있으며, 식품 첨가물로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전청에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다. 상기 '식품 첨가물 공전'에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼슘, 니코틴산, 계피산 등의 화학적 합성물; 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물; L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료 제제, 타르색소제제 등의 혼합제제류 등을 들 수 있다. 예를 들어, 정제 형태의 건강기능식품은 본 발명의 유효성분을 부형제, 결합제, 붕해제 및 다른 첨가제와 혼합한 혼합물을 통상의 방법으로 과립화한 다음, 활택제 등을 넣어 압축성형하거나, 상기 혼합물을 직접 압축 성형할 수 있다. 또한 상기 정제 형태의 건강기능식품은 필요에 따라 교미제 등을 함유할 수도 있다. 캅셀 형태의 건강기능식품 중 경질 캅셀제는 통상의 경질 캅셀에 본 발명의 유효성분을 부형제 등의 첨가제와 혼합한 혼합물을 충진하여 제조할 수 있으며, 연질 캅셀제는 본 발명의 유효성분을 부형제 등의 첨가제와 혼합한 혼합물을 젤라틴과 같은 캅셀기제에 충진하여 제조할 수 있다. 상기 연질 캅셀제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다. 환 형태의 건강기능식품은 본 발명의 유효성분과 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 성형하여 조제할 수 있으며, 필요에 따라 백당이나 다른 제피제로 제피할 수 있으며, 또는 전분, 탈크와 같은 물질로 표면을 코팅할 수도 있다. 과립 형태의 건강기능식품은 본 발명의 유효성분의 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 입상으로 제조할 수 있으며, 필요에 따라 착향제, 교미제 등을 함유할 수 있다.
본 명세서 전체에 걸쳐, 특정 물질의 농도를 나타내기 위하여 사용되는 '%'는 별도의 언급이 없는 경우, 고체/고체는 (w/w) %, 고체/액체는 (w/v) %, 그리고 액체/액체는 (v/v) %이다.
일 구현예에 있어서, 본 발명은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 화장료 조성물을 제공한다.
본 조성물을 첨가할 수 있는 제품으로는, 예를 들어, 각종 크림, 로션, 스킨, 에센스 등과 같은 화장품류와 샴푸, 린스, 클렌징, 세안제, 비누, 트리트먼트, 팩, 미용액 등이 있다.
본 발명의 화장료는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고 지질 및 해초 추출물로 이루어진 군에서 선택된 조성물을 포함한다.
수용성 비타민으로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 비타민 B1, 비타민 B2, 비타민 B6, 피리독신, 염산피리독신, 비타민 B12, 판토텐산, 니코틴산, 니코틴산아미드, 엽산, 비타민 C, 비타민 H 등을 들 수 있으며, 그들의 염 (티아민염산염, 아스코르빈산나트륨염 등)이나 유도체 (아스코르빈산-2-인산나트륨염, 아스코르빈산-2-인산마그네슘염 등)도 본 발명에서 사용할 수 있는 수용성 비타민에 포함된다. 수용성 비타민은 미생물 변환법, 미생물의 배양물로부터의 정제법, 효소법 또는 화학 합성법 등의 통상의 방법에 의해 수득할 수 있다.
유용성 비타민으로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 비타민 A, 카로틴, 비타민 D2, 비타민 D3, 비타민 E (d1-알파 토코페롤, d-알파 토코페롤, d-알파 토코페롤) 등을 들 수 있으며, 그들의 유도체(팔미틴산아스코르빈, 스테아르산아스코르빈, 디팔미틴산아스코르빈, 아세트산dl-알파 토코페롤, 니코틴산dl-알파 토코페롤비타민 E, DL-판토테닐알코올, D-판토테닐알코올, 판토테닐에틸에테르 등) 등도 본 발명에서 사용되는 유용성 비타민에 포함된다. 유용성 비타민은 미생물 변환법, 미생물의 배양물로부터의 정제법, 효소 또는 화학 합성법 등의 통상의 방법에 의해 취득할 수 있다.
고분자 펩티드로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 콜라겐, 가수 분해 콜라겐, 젤라틴, 엘라스틴, 가수 분해 엘라스틴, 케라틴 등을 들 수 있다. 고분자 펩티드는 미생물의 배양액으로부터의 정제법, 효소법 또는 화학 합성법 등의 통상의 방법에 의해 정제 취득할 수 있으며, 또는 통상 돼지나 소 등의 진피, 누에의 견섬유 등의 천연물로부터 정제하여 사용할 수 있다.
고분자 다당으로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 히드록시에틸셀룰로오스, 크산탄검, 히알루론산나트륨, 콘드로이틴 황산 또는 그 염 (나트륨염 등) 등을 들 수 있다. 예를 들어, 콘드로이틴 황산 또는 그 염 등은 통상 포유 동물이나 어류로부터 정제하여 사용할 수 있다.
스핑고 지질로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 세라미드, 피토스핑고신, 스핑고당지질 등을 들 수 있다. 스핑고 지질은 통상 포유류, 어류, 패류, 효모 또는 식물 등으로부터 통상의 방법에 의해 정제하거나 화학 합성법에 의해 취득할 수 있다.
본 발명의 화장료에는 상기 필수 성분과 더불어 필요에 따라 통상 화장료에 배합되는 다른 성분을 배합해도 된다. 이외에 첨가해도 되는 배합 성분으로서는 유지성분, 보습제, 에몰리엔트제, 계면 활성제, 유기 및무기안료, 유기 분체, 자외선 흡수제, 방부제, 살균제, 산화 방지제, 식물 추출물, pH 조정제, 알콜, 색소, 향료, 혈행 촉진제, 냉감제, 제한(制汗)제, 정제수 등을 들 수 있다. 유지 성분으로서는 에스테르계 유지, 탄화수소계 유지, 실리콘계 유지, 불소계 유지, 동물 유지, 식물 유지 등을 들 수 있다.
에스테르계 유지로서는 트리2-에틸헥산산글리세릴, 2-에틸헥산산세틸, 미리스틴산이소프로필, 미리스틴산 부틸, 팔미틴산이소프로필, 스테아르산에틸, 팔미틴산옥틸, 이소스테아르산이소세틸, 스테아르산부틸, 리놀레산 에틸,리놀레산이소프로필, 올레인산에틸, 미리스틴산이소세틸, 미리스틴산이소스테아릴, 팔미틴산이소스테아릴, 미리스틴산옥틸도데실, 이소스테아르산이소세틸, 세바신산디에틸, 아디핀산디이소프로필, 네오펜탄산이소알킬, 트리(카프릴, 카프린산)글리세릴, 트리2-에틸헥산산트리메틸롤프로판, 트리이소스테아르산 트리메틸롤프로판, 테트라2-에틸헥산산펜타엘리슬리톨, 카프릴산세틸, 라우린산데실, 라우린산헥실, 미리스틴산데실, 미리스틴산미리스틸, 미리스틴산세틸, 스테아르산스테아릴, 올레인산데실, 리시노올레인산세틸, 라우린산이소스테아릴, 미리스틴산이소트리데실, 팔미틴산이소세틸, 스테아르산옥틸, 스테아르산 이소세틸, 올레인산이소데실, 올레인산옥틸도데실, 리놀레산옥틸도데실, 이소스테아르산이소프로필, 2-에틸헥산산세토스테아릴, 2-에틸헥산산스테아릴, 이소스테아르산헥실, 디옥탄산에틸렌글리콜, 디올레인산에틸렌글리콜, 디카프린산프로필렌글리콜, 디(카프릴, 카프린산)프로필렌글리콜, 디카프릴산프로필렌글리콜, 디카프린산네오펜 틸글리콜, 디옥탄산네오펜틸글리콜, 트리카프릴산글리세릴, 트리운데실산글리세릴, 트리이소팔미틴산글리세릴, 트리이소스테아르산글리세릴, 네오펜탄산옥틸도데실, 옥탄산이소스테아릴, 이소노난산옥틸, 네오데칸산헥실데실, 네오데칸산옥틸도데실, 이소스테아르산이소세틸, 이소스테아르산이소스테아릴, 이소스테아르산옥틸데실, 폴리글리세린올레인산에스테르, 폴리글리세린이소스테아르산에스테르, 시트르산트리이소세틸, 시트르산트리이소알킬, 시트르산트리이소옥틸, 락트산라우릴, 락트산미리스틸, 락트산세틸, 락트산옥틸데실, 시트르산트리에틸, 시트르산아세틸트리에틸, 시트르산아세틸트리부틸, 시트르산트리옥틸, 말산디이소스테아릴, 히드록시스테아르산 2-에틸헥실, 숙신산디2-에틸헥실, 아디핀산디이소부틸, 세바신산디이소프로필, 세바신산디옥틸, 스테아르산콜레스테릴, 이소스테아르산콜레스테릴, 히드록시스테아르산콜레스테릴, 올레인산콜레스테릴, 올레인 산디히드로콜레스테릴, 이소스테아르산피트스테릴, 올레인산피트스테릴, 12-스테알로일히드록시스테아르산이소세틸, 12-스테알로일히드록시스테아르산스테아릴, 12-스테알로일히드록시스테아르산이소스테아릴 등의 에스테르 계 등을 들 수 있다.
탄화 수소계 유지로서는 스쿠알렌, 유동 파라핀, 알파-올레핀올리고머, 이소파라핀, 세레신, 파라핀, 유동 이소파라핀, 폴리부덴, 마이크로크리스탈린왁스, 와셀린 등의 탄화수소계 유지 등을 들 수 있다.
실리콘계 유지로서는 폴리메틸실리콘, 메틸페닐실리콘, 메틸시클로폴리실록산, 옥타메틸폴리실록산, 데카메틸폴리실록산, 도데카메틸시클로실록산, 디메틸실록산 및 메틸세틸옥시실록산 공중합체, 디메틸실록산 및 메틸스테알록시실록산 공중합체, 알킬 변성 실리콘유, 아미노 변성 실리콘유 등을 들 수 있다.
불소계 유지로서는 퍼플루오로폴리에테르 등을 들 수 있다.
동물 또는 식물 유지로서는 아보카도유, 아르몬드유, 올리브유, 참깨유, 쌀겨유, 새플라워유, 대두유, 옥수수유, 유채유, 행인(杏仁)유, 팜핵유, 팜유, 피마자유, 해바라기유, 포도종자유, 면실유, 야자유, 쿠쿠이너트유, 소맥배아유, 쌀 배아유, 시아버터, 월견초유, 마커데이미아너트유, 메도홈유, 난황유, 우지(牛脂), 마유, 밍크유, 오렌지라피유, 호호바유, 캔데리러왁스, 카르나바왁스, 액상 라놀린, 경화피마자유 등의 동물 또는 식물 유지를 들 수 있다.
보습제로서는 수용성 저분자 보습제, 지용성 분자 보습제, 수용성 고분자, 지용성 고분자 등을 들 수 있다.
수용성 저분자 보습제로서는 세린, 글루타민, 솔비톨, 만니톨, 피롤리돈-카르복실산나트륨, 글리세린, 프로필렌글리콜, 1,3-부틸렌글리콜, 에틸렌글리콜, 폴리에틸렌글리콜B(중합도 n = 2 이상), 폴리프로필렌글리콜 (중합도 n = 2 이상), 폴리글리세린B(중합도 n = 2 이상), 락트산, 락트산염 등을 들 수 있다.
지용성 저분자 보습제로서는 콜레스테롤, 콜레스테롤에스테르 등을 들 수 있다.
수용성 고분자로서는 카르복시비닐폴리머, 폴리아스파라긴산염, 트라가칸트, 크산탄검, 메틸셀룰로오스, 히드록시메틸셀룰로오스, 히드록시에틸셀룰로오스, 히드록시프로필셀룰로오스, 카르복시메틸셀룰로오스, 수용성키틴, 키토산, 덱스트린 등을 들 수 있다.
지용성 고분자로서는 폴리비닐피롤리돈 및 에이코센 공중합체, 폴리비닐피롤리돈 및 헥사데센 공중합체, 니트로셀룰로오스, 덱스트린지방산에스테르, 고분자 실리콘 등을 들수 있다. 에몰리엔트제로서는 장쇄아실글루타민산 콜레스테릴에스테르, 히드록시스테아르산콜레스테릴, 12-히드록시스테아르산, 스테아르산, 로진산, 라놀린지방산콜레스테릴에스테르 등을 들 수 있다.
계면 활성제로서는 비이온성 계면 활성제, 음이온성 계면 활성제, 양이온성 계면 활성제, 양성 계면 활성제 등을 들 수 있다.
비이온성 계면 활성제로서는 자기 유화형 모노스테아르산글리세린, 프로필렌글리콜지방산에스테르, 글리세린 지방산에스테르, 폴리글리세린지방산에스테르, 솔비탄지방산에스테르, POE(폴리옥시에틸렌)솔비탄지방산에스테르, POE 솔비트지방산에스테르, POE 글리세린지방산에스테르, POE 알킬에테르, POE 지방산에스테르, POE 경화피마자유, POE 피마자유, POE 및 POP (폴리옥시에틸렌 및 폴리옥시프로필렌) 공중합체, POE 및 POP 알킬에테르, 폴리에테르변성실리콘, 라우린산알카놀아미드, 알킬아민옥시드, 수소첨가대두인지질 등을 들 수 있다.
음이온성 계면 활성제로서는 지방산비누, 알파-아실술폰산염, 알킬술폰산염, 알킬알릴술폰산염, 알킬나프탈렌술폰산염, 알킬황산염, POE 알킬에테르황산염, 알킬아미드황산염, 알킬인산염, POE 알킬인산업, 알킬아미드 인산염, 알킬로일알킬타우린염, N-아실아미노산염, POE 알킬에테르카르복실산염, 알킬술포숙신산염, 알킬술포아세트산나트륨, 아실화 가수분해 콜라겐펩티드염, 퍼플루오로알킬인산에스테르 등을 들 수 있다.
양이온성 계면 활성제로서는 염화알킬트리메틸암모늄, 염화스테아릴트리메틸암모늄, 브롬화스테아릴트리메틸암모늄, 염화세토스테아릴트리메틸암모늄, 염화디스테아릴디메틸암모늄, 염화스테아릴디메틸벤질암모늄, 브롬화베헤닐트리메틸암모늄, 염화벤잘코늄, 스테아르산 디에틸아미노에틸아미드, 스테아르산디메틸아미노프로필아 미드, 라놀린 유도체 제 4급 암모늄염 등을 들 수 있다. 양성 계면 활성제로서는 카르복시베타인형, 아미드베타인형, 술포베타인형, 히드록시술포베타인형, 아미드 술포 베타인형, 포스포베타인형, 아미노카르복실산염형, 이미다졸린 유도체형, 아미드아민형 등의 양성 계면 활성제 등을 들 수 있다.
유기 및 무기 안료로서는 규산, 무수규산, 규산마그네슘, 탤크, 세리사이트, 마이카, 카올린, 벵갈라, 클레이, 벤토나이트, 티탄피막운모, 옥시염화비스무트, 산화지르코늄, 산화마그네슘, 산화아연, 산화티탄, 산화알루미늄, 황산칼슘, 황산바륨, 황산마그네슘, 탄산칼슘, 탄산마그네슘, 산화철, 군청, 산화크롬, 수산화크롬, 칼라민 및 이들의 복합체등의 무기 안료; 폴리아미드, 폴리에스테르, 폴리프로필렌, 폴리스티렌, 폴리우레탄, 비닐수지, 요소수지, 페놀수지, 불소수지, 규소수지, 아크릴수지, 멜라민수지, 에폭시수지, 폴리카보네이트 수지,디비닐벤젠 및 스티렌 공중합체, 실크파우더, 셀룰로오스, CI 피그먼트옐로우, CI 피그먼트오렌지 등의 유기 안료 및 이들의 무기 안료와 유기 안료의 복합 안료 등을 들 수 있다.
유기 분체로서는 스테아르산칼슘 등의 금속비누; 세틸린산아연나트륨, 라우릴린산아연, 라우릴린산칼슘 등의 알킬인산금속염 ; N-라우로일-베타-알라닌칼슘, N-라우로일-베타-알라닌아연, N-라우로일글리신칼슘 등의 아실아미노산 다가금속염 ; N-라우로일-타우린칼슘, N-팔미토일-타우린칼슘 등의 아미드술폰산 다가금속염 ; N-엡실론-라우로일-L-리진, N-엡실론-팔미토일리진, N-알파-파리토일올니틴, N-알파-라우로일아르기닌, N-알파-경화우지지방산아실아르기닌 등의 N-아실염기성아미노산 ; N-라우로일글리실글리신 등의 N-아실폴리펩티드 ; 알파-아미노카프릴산, 알파-아미노라우린산 등의 알파-아미노지방산 ; 폴리에틸렌, 폴리프로필렌, 나일론, 폴리메틸메타크릴레이트, 폴리스티렌, 디비닐벤젠 및 스티렌 공중합체, 사불화에틸렌 등을 들 수 있다.
자외선 흡수제로서는 파라아미노벤조산, 파라아미노벤조산에틸, 파라아미노벤조산아밀, 파라아미노벤조산옥틸, 살리실산에틸렌글리콜, 살리신산페닐, 살리신산옥틸, 살리신산벤질, 살리신산부틸페닐,살리신산호모멘틸, 계피산벤질, 파라메톡시계피산-2-에톡시에틸, 파라메톡시계피산옥틸, 디파라메톡시계피산모 노-2-에틸헥산글리세릴, 파라메톡시 계피산 이소프로필, 디이소프로필 및 디이소프로필계피산에스테르 혼합물, 우로카닌산, 우로카닌산에틸, 히드록시메톡시벤조페논, 히드록시메톡시벤조페논술폰산 및 그 염, 디히드록시메톡시벤조페논, 디히드록시메톡시벤조페논디술폰산나트륨, 디히드록시벤조페논, 테트라히드록시벤조페논, 4-tert-부틸-4'-메톡시디벤조일메탄, 2,4,6-트리아닐리노-p-(카르보-2'-에틸헥실-1'-옥시)-1,3,5-트리아진, 2-(2-히드록시-5-메틸페닐)벤조트리아졸 등을 들 수 있다.
살균제로서는 히노키티올, 트리클로산, 트리클로로히드록시디페닐에테르, 크로르헥시딘글루콘산염, 페녹시에탄올, 레조르신, 이소프로필메틸페놀, 아줄렌, 살리칠산, 진크필리티온, 염화벤잘코늄, 감광소 301 호, 모노니트로과이어콜나트륨, 운데시렌산 등을 들 수 있다.
산화 방지제로서는 부틸히드록시아니솔, 갈릭산프로필, 엘리소르빈산 등을 들 수 있다.
pH 조정제로서는 시트르산, 시트르산나트륨, 말산, 말산나트륨, 프말산, 프말산나트륨, 숙신산, 숙신산나트륨, 수산화나트륨, 인산일수소나트륨 등을 들 수 있다.
알코올로서는 세틸알코올 등의 고급 알코올을 들 수 있다.
또한, 이외에 첨가해도 되는 배합 성분은 이에 한정되는 것은 아니며, 또, 상기 어느 성분도 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 배합 가능하다.
본 발명의 화장료는 용액, 유화물, 점성형 혼합물 등의 형상을 취할 수 있다.
본 발명의 화장료 조성물에 포함되는 성분은 유효성분으로서 화장료 조성물에 통상적으로 이용되는 성분들을 포함할 수 있으며, 예를 들면, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제 및 담체를 포함한다.
본 발명의 피부 미백 및 피부 노화방지용 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어 유액, 크림, 화장수, 팩, 파운데이션, 로션, 미용액, 모발화장료 등을 들 수 있다.
구체적으로, 본 발명의 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 밀크 로션, 아스트린젠트, 로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 헤어로션, 헤어토닉, 헤어에센스, 헤어샴푸, 헤어린스, 헤어트리트먼트, 바디로션 및 바디클린저의 제형을 포함한다.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화 아연 등이 이용될 수 있다.
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.
본 발명의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.
본 발명의 제형이 계면-활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산디에탄올아미드, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.
일 구현예에 있어서, 본 발명은 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 사료 조성물을 제공한다.
본 발명의 사료 조성물은 기존의 항생제를 대체하고 유해한 식품 병원성균의 생장을 억제하는 바, 동물체의 건강상태를 양호하게 하고, 가축의 증체량과 육질을 개선시키며, 산유량 및 면역력을 증가시키는 효과를 기대할 수 있다. 본 발명의 사료 조성물은 발효사료, 배합사료, 펠렛 형태 및 사일레지 등의 형태로 제조될 수 있다. 상기 발효사료는 본 발명의 펩타이드 이외의 여러 가지 미생물군 또는 효소들을 첨가함으로서 유기물을 발효시켜 제조할 수 있으며, 배합사료는 여러 종류의 일반사료와 본 발명의 펩타이드를 혼합하여 제조할 수 있다. 펠렛 형태의 사료는 상기 배합사료 등을 펠렛기에서 열과 압력을 가하여 제조할 수 있으며, 사일레지는 청예 사료를 본 발명에 따른 미생물로 발효시킴으로써 제조할 수 있다. 습식발효사료는 음식물 쓰레기 등과 같은 유기물을 수집 및 운반하여 살균과정과 수분조절을 위한 부형제를 일정비율로 혼합한 후, 발효에 적당한 온도에서 24시간 이상 발효하여, 수분함량이 약 70%으로 포함되도록 조절하여 제조할 수 있다. 발효건조사료는 습식 발효 사료를 건조과정을 추가로 거쳐 수분함량이 30% 내지 40% 정도 함유되도록 조절하여 제조할 수 있다.
본 발명의 사료 조성물은 종래 사료에 첨가되는 성분을 더 포함할 수 있다. 이러한 사료에 첨가되는 성분의 일예로서 곡류분말, 고기분말, 및 두류 등을 포함할 수 있다. 상기에서 곡류분말은 쌀가루, 밀가루, 보리가루, 및 옥수수가루 중에서 선택된 1종 이상을 사용할 수 있다. 상기에서 고기분말은 닭고기, 소고기, 돼지고기, 및 타조고기 중에서 선택된 어느 하나 이상을 분말화한 고기분말을 사용할 수 있다. 상기에서 두류는 대두, 강낭콩, 완두콩, 및 검정콩 중에서 선택된 1종 이상을 사용할 수 있다.
본 발명의 사료 조성물은 상기에서 언급한 종래 사료에 첨가되는 성분인 곡류분말, 고기분말, 및 두류 이외에도 사료의 영양성을 증대시키기 위해 영양제, 및 무기물 중에서 선택된 어느 하나 이상을 첨가할 수 있으며, 사료 품질의 저하를 막기 위해 항곰팡이제, 항산화제, 항응고제, 유화제, 및 결착제 중에서 선택된 1종 이상을 포함할 수 있다.
이하, 본 발명을 실시예를 통하여 더욱 상세히 설명하기로 한다. 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.
준비예 1. 동물
6주령의 수컷 BALB/c 마우스(21-23g)를 중앙연구소, Animal Inc.(Korea)로부터 구매 하였고, 마우스의 온도는 22 ± 2oC로 유지하였고, 12시간/12시간 암/명 주기을 표준 조건으로 수용되었다.
실험 전 최소 1주일 동안 모든 마우스에게 표준 실험실 식단과 물을 제공하였다. 모든 실험 과정은 강릉원주대학교 위원회의 승인을 받았다.
준비예 2.
고려인삼열매 다당류
한국 충청도 보은에서 구매한 고려인삼열매분말(20 g)을 99% 에탄올로 3 시간 추출하였고, 추출물을 4000 xg에서 15분 동안 원심분리하고 침전물을 실온에서 건조시켰다. 건조된 침전물에 증류수를 첨가하고 100°C에서 3시간 동안 추출하였다. 다당류 추출물은 99% 에탄올을 첨가한 후 원심분리, 여과, 아세톤으로 세척 및 건조된 다당류 침전물에 동결건조하여 조 다당류(GBP-C)를 생성하여 얻었다. GBP-C는 DEAE Sepharose 빠른 흐름 컬럼(17-0709-01; GE Healthcare Bio-Science AB, Uppsala, Sweden)에서 이온 교환 크로마토그래피를 사용하여 분획화되었으며 노스(rhamnose)(4.0%), 아라비노스(arabinose)(19.8%), 만노스(mannose)(2.2%), 글루코스(glucose)(27.7%) 및 갈락토스(galactose)(46.3%)로 구성된 분자량 328.4 및 54.2 × 103(g/mol)의 국산 고려인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 제조하였다.
준비예 3. 면역억제 치료
본 발명에서 40마리의 마우스를 무작위로 8개 그룹(그룹당 n=5)으로 나누어 다양하게 표 1에 따라 처리하였다.
시클로포스파미드(cyclophosphamide, CY)는 80 mg/kg 체중(BW)의 용량으로 복강내(IP) 주사에 의해 면역억제 모델을 유도하는 데 사용하였으며, 레바미솔(Levamisole, LEV)는 세포 매개, 즉 T 세포 유도 면역을 조절할 수 있는 면역 조절제로써 40 mg/kg BW의 용량으로 양성대조군으로 사용되었으며, 상업용 홍삼 시럽100 mg/kg BW의 용량으로 양성 대조군으로 사용하였다. 또한, 인삼은 시중에서 판매되는 제품을 사용하였으며, 표 1의 시료들은 마지막 처리 후 24시간에 마우스를 희생시켰다.
그룹 | 처리(Treatment) | ||
1일 내지 6일(경구) | 4일 내지 6일(IP) | 7일 내지 10일(경구) | |
정상군 | Saline | - | Saline |
CY | Saline | CY | Saline |
홍삼 | 홍삼 시럽 | CY | 홍삼 시럽 |
LEV | 레바미솔 | CY | 레바미솔 |
Tr.1 | GBP(50mg/kg BW) | CY | GBP(50mg/kg BW) |
Tr.2 | GBP(100mg/kg BW) | CY | BP(100mg/kg BW) |
Tr.3 | GBP(250mg/kg BW) | CY | BP(250mg/kg BW) |
Tr.4 | GBP(500mg/kg BW) | CY | BP(500mg/kg BW) |
준비예 4. 비장 지수 측 정
비장을 해부하고 무게를 쟀다. 비장 지수는 하기 계산식에 따라 계산하였다.
[계산식]
비장 지수(mg/g) = 비장 무게/마우스 체중
준비예 5. 마우스 비장 세포 준비
BALB/c 마우스의 비 장에서 비장세포를 분리하기 위해, 비장의 무게를 잰 후, 마우스의 비장을 비장 세포 분리를 위해 얼음처럼 차가운 PBS에 넣었다. 마우스의 비장세포는 RBC Lysis Buffer(eBioscience, USA)를 이용하여 추출하였다. 제조업체 의 지침에 따라, 비장 세포는 1% fetal bovine serum과 1% streptomycin (100 μ g/ml), penicillin (100 IU/ml) (Welgene, Korea)이 첨가된 RPMI1640 배지에 (Gibco Laboratories, USA)에서 2 × 106 cells/ml의 농도로 조정되었다.
준비예 6. 비장 세포 증식 측정
비장 림프구 증식을 측정하기 위해 EZ-Cytox Cell Viability Assay Kit(대일랩 서비스, 한국)를 사용하였다.
비장 세포를 96-웰 플레이트에 시딩하고 37℃, 5% CO2 에서 1시간 동안 배양하였다. 배양한 비장 세포를 T 세포분열촉진제 로서 5 μg/ml 콘카나발린 A(Con A) 또는 B 세포 세포분열촉진제로서 10 μg/ml 리포폴리사카라이드(LPS)로 자극하였다. 37°C에서 5% CO 2로 48시간 동안 인큐베이션한 후, 25 μl EZ-Cytox 시약을 각 웰에 첨가하고 플레이트를 37°C에서 5% CO2로 1 시간 추가로 인큐베이션하였다. 마 이크로플레이트 리더(EL800; BioTek, USA)를 사용하여 450 nm에서 흡광도를 측정하 였다.
준비예 7. NK 세포 매개 세포 독성 평가 분석
비장 세포는 YAC-1 세포(한국 세포주 은행)와 함께 배양 하였거나, YAC-1 세 포 없이 공동 배양되어 이펙터 세포 대 NK 민감성 타겟 세포의 비율을 50:1로 얻었다. 24시간 배양 후, 원심분리 후 상층액을 모았다. 250 ×g에서 5분 동안 NK 세포 의 활성은 CytoTox 96 Non-Radioactive Cytotoxicity Assay로 키트(프로메가, 미 국)의 평가하였다. 그 다음 490 nm에서의 흡광도를 측정하였다.
준비예 8.
정량적 RT-PCR(qRT-PCR) 분석
비장 림프구에서 총 RNA를 추출하기 위해 Tri 시약 (Molecular Research Center, Inc., USA)을 사용하였 다. 총 RNA는 고용량 cDNA 역전사 키트(Applied Biosystems, USA)를 사용하여 cDNA로 역전사 하였다. cDNA를 증폭하기 위해 QuantStudio 3 Flex Real-Time PCR System(Thermo Fisher Scientific, USA)에서 TB Green Premix Ex Taq II(Takara Bio Inc., 일본)를 사용하여 cDNA를 qRT-PCR에 적 용하였다. 증폭 조건은 다음과 같다: 95°C에서 30초 동안 초기 변성, 이어서 95° C에서 5초 및 60°C에서 34초의 40주기. 프라이머 서열은 표 2에 나열하였다.
유전자 | Accession No. | 프라이머 서열(5'->3') |
IL-1β | NM_008361.4 | Forward: GGGCCTCAAAGGAAAGAATC |
Reverse: TACCAGTTGGGGAACTCTGC | ||
IL-2 | NM_008366.3 | Forward: CCTGAGCAGGATGGAGAATTACA |
Reverse: TCCAGAACATGCCGCAGAG | ||
IL-4 | NM_021283.2 | Forward: ACAGGAGAAGGGACGCCAT |
Reverse: GAAGCCCTACAGACGAGCTCA | ||
IL-6 | NM_031168.2 | Forward: AGTTGCCTTCTTGGGACTGA |
Reverse: CAGAATTGCCATTGCACAAC | ||
TNF-α | D84199.2 | Forward: ATGAGCACAGAAAGCATGATC |
Reverse: TACAGGCTTGTCACTCGAATT | ||
IFN-γ | NM_008337.3 | Forward: CTCAAGTGGCATAGATGT |
Reverse: GAGATAATCTGGCTCTGCAGGATT | ||
TLR-4 | NM_021297.3 | Forward: CGCTCTGGCATCATCTTCAT |
Reverse: GTTGCCGTTTCTTGTTCTTCC | ||
COX-2 | NM_011198.4 | Forward: AGAAGGAAATGGCTGCAGAA |
Reverse: GCTCGGCTTCCAGTATTGAG | ||
β-actin | NM_007393.5 | Forward: CCACAGCTGAGAGGGAAATC |
Reverse: AAGGAAGGCTGGAAAAGAGC |
준비예 9. 통계 분석
통계분석은 일원 분산 분석(ANOVA)과 Statistix 8.1 통계 소프트웨어(Statistix, USA) 를 사용한 Tukey의 사후 테스트를 사용하여 평가했습니다. p-값이 0 .05 미만일 때 처리군 간의 유의차가 있는 것으로 간주하였다.
실시예 1. 고려 인삼 열매 다당류(GBP)의 비장 지수의 영향
고려 인삼 열매 다당류 (GBP)의 비장 크기 및 비장 지표에 미치는 영향을 확인하기 위해, 시클로포스파미드(cyclophosphamide, CY)(80 mg/kg)을 마우스 투여하여 면역 억제된 마우스에 레바미솔(Levamisole, LEV)(40 mg/kg) 처리군, 홍삼(100 mg/kg) 처리군, 고려 인삼 열매 다당류(GBP) 농도별(50, 100, 250 및 500 mg/kg BW)로 처리한 군에 비장 크기 및 비장 지표를 확인 하였다.
우선, 정상군과 비교하여 사 클로포스파미드(CY) 처리군은 양쪽 비장 크기 및 비장 크기 및 지표 는 감소하였으며, 레바미솔(LEV) 처리군, 홍삼 처리군은 증가된 비장 크기 및 지표를 확인 하였으며, 고려 인삼 열매 다당류(GBP) 처리군은 농도 의존적으로(50, 100, 250 및 500 mg/kg BW) 증가된 비장 크기 및 지표를 확인 하였다.
따라서, 본 발명의 고려 인삼 열매 다당류(GBP)는 비장 크기(도 1) 및 비장 지표(도 2)를 증가시켜 면역을 증가시키는 것을 시사한다.
실시예 2. 고려 인삼 열매 다당류(GBP)의 비장 림프구 증식의 영향
고려 인삼 열매 다 당류(GBP)의 비장 림프구 증식에 미치는 영향을 확인하기 위해, 비 장 세포를 T 세포분열촉진제로서 분열 촉진제 콘카나발린 A(concanavalin A, Con A) 또는 B 세포 세포분열촉진제로서 리포폴리사카라이드(lipopolysaccharide, LPS)로 자극 시켜, 비장 림프구 증식에 대한 고려 인삼 열 매의 다당류(GBP)의 효과를 확인하였다.
시클로포스파미드(CY) 처리군의 콘카나발린 A( Con A) 또는 리포폴리사카라이드(LPS)로 자극된 세포 증식(각각 48 .1% 및 57.1%)이 정상군에 비해 현저히 감소한 것을 확인 하였다.
고려 인삼 열매 다당류(GBP)의 250 및 500 mg/kg BW 농도에서 림프구의 증식 을 상당히 자극하였고, 시클로포스파미드(CY) 처리군과 비교하여 콘카나발린 A(Con A) 또는 리포폴리사카라이드(LPS)에 의해 유도된 비장 세포는 고려 인삼 열매 다당 류(GBP)의 500 mg/kg BW 농도에서 림프구 증식이 양성대조군인 홍삼 처리군 및 렘 바미솔(LEV)처리군 보다 T 림프구 및 B 림프구의 높은 증식을 확인하였다(도 3).
따라서 , 본 발명의 고려 인삼 열매 다당류(GBP)는 T 림프구 및 B 림프구의 증식을 증가시켜 면역을 증가시키는 것을 시사한다.
실시예 3 . 고려 인삼 열매 다당류(GBP)의 NK 세포 활성에 미치는 영향
고려 인삼 열매 다당 류(GBP)의 NK 세포 활성에 미치는 영향을 확인하기 위해, 시클로포스파미드(CY) 처리군은 NK 세포의 활성을 42.9%로 상당히 억제되었다.
반면, 고려 인삼 열매 다당류(GBP)의 100, 250, 500 mg/kg BW 농도에서 NK 세포 활성 은 농도 의존적으로 시클로포스파미드(CY) 처리군보다 유의하게 높은 것 을 확인하였다.
또한 고려 인삼 열매 다당류(GBP) 500 mg/kg BW 처리 시, 비장 NK 세포 활성이 정상군과 유사한 수준으로 회복을 확인 하였다(도 4).
따라서, 본 발명의 고려 인삼 열매 다당류(GBP)는 NK 세포 활성을 증가시켜 면역을 증가시키는 것을 시사한다.
실시예 4. 고려 인삼 열매 다당류(GBP)의 면역 관련 유전자 발현에 미치는 영향.
고려 인삼 열매 다당류(GBP)의 면역 관련 유잔자 발현에 미치는 영향을 확인 하기 위해, 고려 인삼 열매 다당류(GBP)가 T 세포분열촉진제로서 분열 촉진제 콘카 나발린 A(Con A) 또는 B 세포 세포분열촉진제로서 리포폴리사카라이드(LPS)에 의해 자극된 비장 림프구에서 면역 반응을 자극할 수 있는지 여부를 평가하기 위해 실시 간 PCR을 수행하여 IL-1β와 같은 면역 관련 유전자의 mRNA 발현 수준을 조사하였 다.
면역 관련 유전자인 IL-2, IL-4, IL-6, TNF-α, IFN-γ, TLR-4 및 COX-2 mRNA 시클로포스파미드(CY) 처리군에서 면역 관련 유전자의 발현 수준은 정상군 보 다 낮았다.
모든 고려 인삼 열매 다당류(GBP)처리군은 (50-500 mg/kg BW)은 시클로포스 파미드(CY) 처리군 보다 면역 관련 유전자의 발현 수준이 현저하게 증가하는 것을 확인 하였다. 고려 인삼 열매 다당류(GBP)처리군은 I L-1β, IL-2 및 IL-6의 발현 수준은 B 세포 세포분열촉진제로서 리포 폴리사카라이드(LPS) 보다 T 세포분열촉진제로서 분열 촉진제 콘카나 발린 A(Con A)의해 유의하게 상향 조절되는 것을 확인 하였다(도 5, 도 6, 도 8).
반면, IL-4, TNF-α, IFN-γ, TLR-4 및 COX-2 발현 수준 T 세포분열촉진 제 보다 B 세포 세포분열촉진제로서 리포폴리사카라이드(LPS)에 의 해 상당히 상향조절되는 것을 확인 하였다(도 7, 도 9, 도 10, 도 11, 도 12).
고려 인삼 열매 다당류(GBP)(250 및 500mg/kg BW)에서 정상 그룹보다 면역관 련 유전자의 발현 수준을 증가시키는 것을 확인 하였다.
따라서, 본 발명의 고려 인삼 열매 다당류(GBP)는 면역관련유전자를 증가시켜 면역을 증가시키는 것을 시사한다.
고려 인삼 열매 다당류(GBP)는 시클로포스파미드 (CY)는 유도 면역억제 마우스에서 강력한 면역조절 특성을 나타내며 , 면역억제된 마우스의 고려 인삼 열매 다당류(GBP)는 치료는 비장 지수를 개선하여 면역 반응을 강화하였다.
또한, 비장 세포 증식 및 NK 세포 활성 면역 조절 사이토카인의 유전자 발현을 상향 조절하였다. 고려 인삼 열매 다당류(GBP)는 생체 내 마우스 모델에서 면역 강화 활성을 가지고 있 음을 보고한 최초의 발명이며, 고려 인삼 열매(GBP)가 효과적인 면역 조절제로 개발될 수 있음을 시사한다.
Claims (13)
- 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 식품 조성물.
- 제 1항에 있어서,상기 고려 인삼 열매 추출물은 물, C1 내지 C4의 저급 알코올 또는 이들의 혼합용매로 이루어 진 군에서 선택된 어느 하나의 용매로 추출된 것인, 조성물.
- 제 1항에 있어서,상기 고려 인삼 열매 다당류는 람노스(rhamnose), 아라비노스(arabinose), 만노스(mannose), 글루코스(glucose) 및 갈락토스(galactose)로 이루어진 것인, 조성물.
- 제 1항에 있어서,상기 고려 인삼 열매 다당류의 분자량 328.4 및 54.2 × 103(g/mol)으로 구성하는 것인, 조성물.
- 제 3항에 있어서,상기 고려 인삼 열매 다당류는 람노스의 4.0%, 아라비노스는 19.8%, 만노스는 2.2%, 글루코스 27.7% 및 갈락토스는 46.3%로 포함된 것인, 조성물.
- 제 1항에 있어서,상기 조성물은 비장의 크기 및 비장 지표를 증가시키는 것인, 조성물.
- 제 1항에 있어서,상기 조성물은 T 림프구 및 B 림프구를 증가시키는 것인, 조성물.
- 제 1항에 있어서,상기 조성물은 NK 세포 활성을 증가시키는 것인, 조성물.
- 제 1항에 있어서,상기 조성물은 면역 관련 유전자를 증가 시키는 것인, 조성물.
- 제 9항에 있어서,상기 면역 관련 유전자는 IL-2, IL-4, IL-6, TNF-α, IFN-γ, TLR-4 및 COX-2로 이루어진 군에서 선택되는 것인, 조성물.
- 제 10항에 있어서,상기 면역은 항암제, 면역억제제 또는 감염에 의한 면역이 저하된 것인, 조성물.
- 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 화장료 조성물.
- 고려 인삼 열매 다당류(Korean ginseng berry polysaccharides, GBP)를 유효성분으로 포함하는 면역증강용 사료 조성물.
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KR1020220176812A KR20240094494A (ko) | 2022-12-16 | 면역저하상태에서의 인삼 열매 다당류의 선천성 면역조절 효과 |
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