Food material with excellent immunoregulatory effect and food or drink product containing the same

Abstract

To provide a food material with an excellent immunoregulatory effect and a food or drink product containing the same, expected to prevent / suppress / reduce inflammation, having an active ingredient that is a safe food-derived ingredient with little side effects.SOLUTION: The food material with the excellent immunoregulatory effect for adjusting a cytokine level of a cell contains polyphenol and a polyphenol oxidative enzyme.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a food material having an excellent immunomodulatory effect and a food or drink containing the same. [Background technology]

[0002] The immune system, along with the nervous system and endocrine system, is an important system for maintaining homeostasis in the body. The immune system is a biological system primarily recognized as a defense against foreign substances, but its function is closely related to various other biological systems, and disturbances in the immune system are involved in various diseases. Therefore, properly regulating the immune system is important for maintaining health, and there is hope for the development of materials with excellent immune-regulating effects. The immune system is maintained by various cells and biological molecules, among which cytokines are important factors. Cytokines are small proteins secreted by cells that play an important role in maintaining the balance of the immune system, such as by transmitting information between cells and activating or suppressing immune cells. Therefore, excellent cytokine-regulating effects are important in expecting immunomodulatory effects. Among cytokines, those produced to promote inflammation in the body play an important role in promoting the removal of foreign substances and the repair of damaged areas. However, excessive or prolonged production of such cytokines contributes to the exacerbation of many chronic diseases, including obesity, diabetes, bronchitis, pancreatitis, colitis, and cardiovascular disease. Therefore, the development of materials that can moderately regulate excessive cytokine production is particularly desirable. While several drugs have been developed to date, their long-term administration may cause side effects. Therefore, there has been growing interest in substances derived from natural products that can be incorporated into the diet. However, such substances are only available in small amounts in foods, which may not be sufficient to elicit the desired effects. In this context, methods to enhance effects by combining multiple materials or substances may be useful, and from this perspective, there has been a desire to develop materials with excellent immune-modulating and cytokine-modulating effects. Furthermore, it has been hoped that such materials would be easy to incorporate into daily life, safe, secure, and easily manufactured. WO 2022 / 138698 shows that ingestion of an agent containing polyphenols and polyphenol oxidase as active ingredients improves the intestinal flora, and describes that it can be used as an agent for the prevention or treatment of inflammatory bowel disease and allergies. However, its immunomodulatory function has not been clarified. JP 2014-523883 A describes a system in which a conjugate of a phenolic compound and hydrogen peroxide is activated by an oxidase expressed in response to tissue damage, resulting in target-site-specific physiological activity. The system demonstrates antibacterial activity and toxin (LPS / cholera toxin) inactivation activity, and the system can be used as an antitoxin, anti-inflammatory, and antibacterial agent. However, there is no mention of the effect of combining polyphenols with polyphenol oxidase to regulate cytokine levels in cells. Furthermore, the agent, which contains hydrogen peroxide, cannot be safely taken orally. Chinese Patent Application Publication No. 114524874 discloses a method for polymerizing ovalbumin (OVA) and destroying epitopes using caffeic acid, laccase, and galactomannan to reduce the allergenicity of OVA, and also describes the application of the resulting hypoallergenic high-molecular OVA to therapeutic drugs. However, it is not known that the combination of polyphenols and polyphenol oxidase has the effect of regulating the amount of cytokines in cells. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2022 / 138698 [Patent Document 2] Special Publication No. 2014-523883 [Patent Document 3] Chinese Patent Application Publication No. 114524874 Summary of the Invention [Problem to be solved by the invention]

[0004] The present invention aims to provide a food material that has excellent immunomodulatory effects and contains safe food-derived ingredients with few side effects, which are expected to have the effects of preventing, suppressing, and reducing inflammation, as well as foods and beverages containing the same. [Means for solving the problem]

[0005] The present invention relates to the following [1] to [9]. [1] A food material containing polyphenols and polyphenol oxidase as active ingredients, which has excellent immunomodulatory effects for regulating the amount of cytokines in cells. [2] The food material according to [1], wherein the polyphenols include o-diphenol polyphenols and / or p-diphenol polyphenols. [3] The food material according to [1] or [2], wherein the polyphenols include polyphenols derived from a polyphenol-containing plant or an extract thereof. [4] The food material according to any one of [1] to [3] above, wherein the polyphenol oxidase comprises a polyphenol oxidase derived from a polyphenol oxidase-containing plant or an extract thereof. [5] The food material described in any one of [1] to [4], wherein regulating the amount of cytokines includes negatively regulating the increase in the amount of cellular inflammatory cytokines induced by stimulation with pathogen-associated molecular patterns (PAMPs). [6] The food material according to [5], which regulates the amount of inflammatory cytokines in cells when the pathogen-associated molecular pattern (PAMP) is LPS. [7] The food material described in [6] above, wherein the inflammatory cytokine is selected from the group consisting of IL-6, TNF-α, and IL-1β. [8] The food material according to any one of [1] to [7] above, wherein the cells include monocyte cells. [9] A food or drink for immune regulation, comprising the food material according to any one of [1] to [8] above. [Effects of the Invention]

[0006] According to the present invention, it is possible to provide a food material having excellent immunomodulatory effects, which is expected to have the effect of preventing, suppressing, and reducing inflammation and which contains safe food-derived components with few side effects as active ingredients, as well as foods and beverages containing the same. DETAILED DESCRIPTION OF THE INVENTION

[0007] The polyphenols contained in the food material of the present invention refer to all compounds having a polyphenol structure in which at least two hydroxy groups are bound to the same benzene ring. The polyphenols also include glycosides. The polyphenols may be used alone or in combination of two or more. Specific examples of the polyphenols include apigenin, apigenin glycoside, acacetin, isorhamnetin, isorhamnetin glycoside, isoquercitrin, epicatechin, epicatechin gallate, epigallocatechin, epigallocatechin gallate, esculetin, ethylprotocatechuate, ellagic acid, catechol, gamma acid, catechin, gardenin, gallocatechin, caffeic acid, caffeic acid ester, chlorogenic acid, kaempferol, kaempferol glycoside, quercetin, quercetin glycoside, quercetagenin, genisetin, and genisetin glycoside. , gossypetin, gossypetin glycoside, gossypol, 4-dihydroxyanthraquinone, 1,4-dihydroxynaphthalene, cyanidin, cyanidin glycoside, sinensetin, diosmetin, diosmetin glycoside, 3,4'-diphenyldiol, sinapinic acid, stearyl-β-(3,5-di-t-butyl-4-hydroxyphenyl)propionate, spinacetin, tangeretin, taxifolin, tannic acid, daphnetin, tyrosine, delphinidin, delphinidin glycoside, theaflavin, theaflavin monogallate, theaflavin bisgallate , Tricetinidine, L-Dopa, Dopamine, Naringenin, Naringin, Nordihydroguaiaretic Acid, Noradrenaline, Hydroquinone, Vanillin, Patchuletin, Herbacetin, Vanillyl Alcohol, Vanitrop, Vanillin Propylene Glycol Acetal, Vanillic Acid, Bis(4-hydroxyphenyl)sulfonic Acid, Bisphenol A, Pyrocatechol, Vitexin, 4,4'-Biphenyldiol, 4-t-Butylcatechol, 2-t-Butylhydroquinone, Protocatechuic Acid, Phloroglucinol, Phenolic Resin, Prussian Nidin, prodelphinidin, phloretin, phloretin glycoside, fisetin, folin, felbacetin, fraxetin, phloridzin, peonidin, peonidin glycoside, perorgonidin, perargonidin glycoside, petunidin, petunidin glycoside, hesperetin, hesperedin, gallic acid, gallic acid esters (methyl gallate, ethyl gallate, dodecyl gallate, lauryl gallate, propyl gallate, butyl gallate, octyl gallate, etc.), mangiferin, malvidin, malvidin glycoside, myricetin, myricetin glycoside, 2,Examples of such compounds include 2'-methylenebis(4-methyl-6-t-butylphenol), 2,2'-methylenebis(4-ethyl-6-t-butylphenol), 2,2'-methylenebis(4-methyl-6-t-butylphenol), 2,2'-methylenebis(4-ethyl-6-t-butylphenol), methyl atralate, 4-methylcatechol, 5-methylcatechol, 4-methoxycatechol, 5-methoxycatechol, methylcatechol-4-carboxylic acid, 2-methylresorcinol, 5-methylresorcinol, morin, limocitrin, limocitrin glycoside, limocitrol, luteolin, luteolin glycoside, luteolinidin, luteolinidin glycoside, rutin, resorcin, resveratrol, resorcinol, leucocyanidin, leucodelphinidin, and rosmarinic acid. Among these polyphenols, flavonoids such as quercetin, epicatechin, and epigallocatechin and their glycosides, o-diphenol polyphenols and / or p-diphenol polyphenols such as gallic acid, gallic acid esters, chlorogenic acid, caffeic acid, caffeic acid esters, tannic acid, pyrocatechol, nordihydroguailectic acid, L-dopa, 4-methylcatechol, 5-methylcatechol, 4-methoxycatechol, and 5-methoxycatechol, and hydroquinone are preferred, and chlorogenic acids or catechins are particularly preferred from the viewpoints of ease of handling, availability, and activity when combined with polyphenol oxidase. The polyphenols can be prepared by known methods, but they may also be commercially available or synthesized. In the present invention, the polyphenols are preferably derived from polyphenol-containing plants or extracts thereof. The term "polyphenol-containing plants" refers to the plant itself, i.e., its leaves, flowers, fruits, stems, skin, and (bulb) roots. Examples of such polyphenol-containing plants include dried products obtained by drying the leaves, flowers, fruits, stems, skin, and (bulb) roots, pulverized (powders) of the dried products, and raw, undried pulverized (powders). The term "polyphenol-containing plant extracts" refers to liquids obtained by extracting the plant with water, e.g., distilled water or ion-exchanged water, or with a hydrophilic or hydrophobic organic solvent, or (freeze-dried) extracts. The extraction method is not limited to solvent extraction, and supercritical extraction and other methods are also acceptable. The extracts used may be those prepared by known methods or commercially available products. The type of polyphenol-containing plant is not particularly limited as long as it does not impair the intended object of the present invention.Examples of polyphenol-containing plants in the present invention include aloe, anise seed, elder, eleutherococcus, plantain, orange flower, allspice, oregano, valerian, chamomile, capsicum pepper, cardamom, cassia, garlic, caraway seed, clove, cumin seed, cola, coriander seed, oak gallnut, saffron, Japanese pepper, juniper berry, cinnamon, ginger, star anise, St. John's Walnut, celery seed, sesame, rhubarb, tarragon, and turmeric. Coffee, teasel, daleseed, nutmeg, nettle, hibiscus, hamamelis, birch, basil, bitter orange, fennel, primrose, fenugreek, verbena, bay laurel, hops, Bordeaux, horseradish, poppy seeds, gall nuts, marigold, mallow, marjoram, mustard, milfoil, mint leaves, melissa, mace, linden, gentian, rose hips, rosemary, rosemary, sunflower seeds, grape skins, apples, carrot leaves, bananas, strawberries, apricots, peaches, plums, pies Napple, pear, persimmon, cherry, papaya, mango, avocado, melon, loquat, fig, kiwi, prune, blueberry, blackberry, raspberry, cranberry, coffee (raw) beans, cocoa (raw) beans, grape seeds, grapefruit seeds, pecan nuts, cashew nuts, chestnuts, coconut, peanuts, walnuts, green tea leaves, black tea leaves, oolong tea leaves, kuding tea, yerba mate tea, rooibos tea, tobacco, perilla leaves, snow thyme, sage, lavender, spearmint, peppermint, santorini, hyssop, basil, marigold Examples of herbs that may be used include dandelion, artichoke, German chamomile, agrimony, licorice, anise, yarrow, eucalyptus, wormwood, balm, angelica, fenugreek, shishito pepper, fennel, chili pepper, coriander seed, caraway seed, fennel seed, ginger, horseradish, marjoram, oregano, Japanese mint, mustard greens, parsley, pepper, savory, tarragon, turmeric, wasabi, dill seed, citrus fruits, pear, thyme, and all kinds of vegetables such as carrots, burdock, bell peppers, turnips, and potatoes.Particularly preferred are rosemary, sunflower seeds, grape skins, apples, carrot leaves, green coffee beans, green cacao beans, grape seeds, green tea leaves, black tea leaves, oolong tea leaves, perilla leaves, snow thyme, sage, spearmint, peppermint, pears, bananas, thyme, Chinese gallnuts, and gall nuts. Green coffee beans and various teas are particularly preferred in terms of ease of handling, availability, and activity when combined with polyphenol oxidase. The polyphenol-containing plant may be a single plant or a mixture of multiple plants.

[0008] The polyphenol oxidase contained in the food material of the present invention is an enzyme that has the ability to oxidize the above-mentioned polyphenols to compounds having a quinone structure, or an enzyme that has the ability to oxidize the polyphenols to quinones by adding a phenolic hydroxyl group to the polyphenols. Examples of the polyphenol oxidase include, but are not limited to, catechol oxidase, polyphenol oxidase, tyrosinase, laccase, glucose oxidase, peroxidase, and ascorbic acid oxidase. The polyphenol oxidase can be prepared by known methods, but commercially available products may also be purchased. Polyphenol oxidases obtained from plants or fungi can also be used. In the present invention, it is preferable to use a crude enzyme (polyphenol oxidase) derived from a plant. That is, the polyphenol oxidase is preferably derived from a polyphenol oxidase-containing plant or an extract thereof. The polyphenol oxidase-containing plant refers to the leaves, flowers, fruits, stems, skin, and (bulb) roots of the plant. Examples of the polyphenol oxidase-containing plant include (freeze)-dried products obtained by (freeze) drying the leaves, flowers, fruits, stems, skin, and (bulb) roots, as well as pulverized (powders) of the (freeze)-dried products, and raw, undried pulverized (powders). More specifically, the plants of the present invention include freeze-dried powders and hot-air-dried powders obtained by freeze-drying these plants. Furthermore, the plants of the present invention also include those obtained by pulverizing these plants using a mixer, immersing them in an organic solvent such as acetone or alcohol, filtering, and vacuum-drying them to obtain a powder. Furthermore, the term "extract of a plant containing polyphenol oxidase" refers to a liquid obtained by extracting the plant with water, such as distilled water or ion-exchanged water, or with a hydrophilic or hydrophobic organic solvent, or a (freeze-)dried extract. The extract may be prepared by a known method or may be commercially available. Since the enzymatic activity of polyphenol oxidase, like that of other enzymes, decreases over time when exposed to heat, it is necessary to use a method that minimizes heat damage to the enzyme (e.g., conventional heat sterilization or sterilization) during the process of obtaining the extract. The polyphenol oxidase-containing plants and polyphenol oxidase-containing fungi are not particularly limited as long as they do not impair the intended purpose of the present invention. Examples of the polyphenol oxidase-containing fungi include mushrooms of the genus Agaricus, such as mushrooms, and mushrooms of the genus Boletus, such as Polylutinis. Examples of the polyphenol oxidase-containing plants include apples, bananas, pears, European pears, strawberries, persimmons, pineapples, grapes, apricots, peaches, plums, papayas, quince, avocados, mangoes, cherries, apricots, melons, loquats, figs, prunes, kiwis, blueberries, blackberries, raspberries, cranberries, currants, burdock, eggplants, tomatoes, mugwort, lotus roots, lettuce, cabbage, and the like. These include sugar beet, hops, barley snips, spinach, radish, turnip, cauliflower, chicory, onion, celery, carrot, asparagus, horseradish, ginger, aloe, bell pepper, barley, wheat, corn, alfalfa, malt, broad beans, soybeans, adzuki beans, green beans, string beans, mung beans, potatoes, sweet potatoes, sugarcane, taro, tea, tobacco, olives, nutmeg, and chrysanthemums. In the present invention, the polyphenol oxidase-containing plant is preferably burdock, prune, pear, European pear, or apple from the viewpoints of ease of handling, availability, and strength of polyphenol oxidase activity. The polyphenol-containing plant may be one species or a mixture of multiple species.

[0009] The food material of the present invention is intended to regulate the amount of (inflammatory) cytokines in cells. A food material with excellent immunomodulatory effects refers to a food material that regulates abnormal immune responses. It also includes agents that maintain and enhance normal immune function. The food material also includes food materials that exert immunomodulatory functions by directly or indirectly acting on cells or tissues involved in immune responses. The food material also includes food materials with excellent (inflammatory) cytokine-regulating effects, food materials with excellent (inflammatory) cytokine production-regulating effects, and food materials with excellent (inflammatory) cytokine gene expression-regulating effects. The food material of the present invention regulates (inflammatory) cytokine gene expression levels, but also has excellent effects on regulating various intracellular events upstream and downstream of (inflammatory) cytokine gene expression in intracellular signaling pathways, as well as various immune-related molecules. For example, the food material of the present invention regulates the expression levels of inflammatory cytokine genes that increase in response to LPS stimulation of cells, but also regulates any intracellular events or immune-related molecules that occur between LPS stimulation and inflammatory cytokine gene expression, and also regulates downstream (inflammatory) cytokine production levels. Furthermore, the food material also provides the function of regulating cytokines other than inflammatory cytokines by regulating various intracellular events and various immune-related molecules. In the present invention, the term "cells" refers to cells that produce cytokines, and includes almost all biological cells, such as immune cells such as macrophages, neutrophils, eosinophils, monocytes, B lymphocytes, T lymphocytes, mast cells, and dendritic cells, as well as endothelial cells, epithelial cells, and fibroblasts. The cells are preferably monocytes. Cytokines are small proteins secreted by cells. They include chemokines, interferons (IFNs), interleukins (ILs), and tumor necrosis factors (TNFs). Cytokines secreted by cells act on the cell itself, as well as on nearby or distant cells, tissues, and molecules, regulating their activity. Various cytokines promote or suppress events within the body and within cells, forming a complex immune regulatory network. Proinflammatory cytokines are cytokines whose production is stimulated by external factors such as microorganisms, viruses, and parasites, or by internal factors released from damaged cells and tissues. Proinflammatory cytokines also include cytokines whose production is stimulated by cytokines from other cells. Each cytokine exerts diverse physiological effects and works cooperatively to promote the attraction, proliferation, activation, and differentiation of immune cells, creating local or systemic inflammation that promotes the elimination of external factors and the repair of self-tissues. Examples of inflammatory cytokines include IL-1, IL-6, IL-8, IL-11, IL-17, IL-18, IFN, and TNF-α. Inflammatory cytokines are important biomolecules for eliminating external factors and repairing autologous tissues. However, chronically enhanced or transiently enhanced overproduction of inflammatory cytokines can have negative effects on biological tissues. For example, inflammatory cytokines are involved in many chronic diseases, including obesity, diabetes, bronchitis, pancreatitis, colitis, cardiovascular disease, sepsis, and gingivitis. The food material of the present invention regulates the levels of inflammatory cytokines, particularly IL-6, TNF-α, and IL-1β. That is, the food material of the present invention regulates the production and gene expression levels of inflammatory cytokines, particularly IL-6, TNF-α, and IL-1β. Modulating cytokine levels can also mean negatively regulating the increase in cellular inflammatory cytokine levels induced by pathogen-associated molecular patterns (PAMPs). Pathogen-associated molecular patterns (PAMPs) are molecular patterns conserved in certain microorganisms and can be recognized by cellular Toll-like receptors (TLRs) and other pattern recognition receptors (PRRs). When cells recognize PAMPs via receptors, various intracellular immune-related molecules are activated or regulated, acting to protect the cells from pathogens and other threats that may threaten their activity. Examples of activated intracellular immune-related molecules include myd88, TRIF, TRAF, IRAK, and NF-κB. The activation or regulation of these intracellular immune-related molecules then results in the regulation of gene expression of various cytokines. Various types of molecules function as PAMPs, including LPS (recognized by TLR4), flagellin (recognized by TLR5), peptidoglycan (recognized by TLR2), virus-associated double-stranded RNA (recognized by TLR3), and CpG (recognized by TLR9). The food material of the present invention regulates the levels of cellular inflammatory cytokines, particularly when the pathogen-associated molecular pattern (PAMP) is LPS. Bacterial lipopolysaccharide (LPS), an endotoxin present on the cell membrane of Gram-negative bacteria, is one PAMP that is specifically recognized by TLR4, a recognition receptor of the innate immune system, to activate or regulate intracellular immune-related molecules and enhance or suppress the production of various cytokines. Representative cytokines that are enhanced include IL6, IL-1β, and TNF-α.

[0010] Examples of foods and drinks for immune regulation that contain the food material of the present invention include liquid products such as fruit juice, fruit juice drinks, non-fruit juice drinks, vegetable drinks, carbonated drinks, sports drinks, coffee drinks, tea, black tea, oolong tea, mineral drinks, energy drinks, alcoholic drinks, non-alcoholic drinks, soups, and noodle soup; candy, chewing gum, tablets, gummies, jelly, chocolate, baked goods such as cookies and cakes, cotton candy, bread, ice cream, frozen desserts, ham, sausages, snacks, seasonings such as powdered sauces; oils and fats such as butter and margarine; solid products such as edible sheet foods; and semi-solid or fluid products such as curry, stew, hayashi rice, sauces, dressings, fresh cream, cream, jam, and liquid foods.

[0011] The food and beverage products may contain various compounding agents or additives commonly used in these products, such as antioxidants, preservatives, antibacterial agents, pH adjusters, sweeteners, acidulants, bulking agents, colorants, emulsifiers, functional substances, flavor improvers, milk components, nitrogen-containing compounds such as amino acids and peptides, various flavor materials, surfactants, abrasives, binders, humectants, and other solvents. Two or more of these compounding agents or additives may be used in any combination. More specifically, examples of sweeteners include sugar, fructose, lactose, glucose, palatinose, maltose, trehalose, sorbitol, erythritol, maltitol, reduced palatinose, xylitol, lactitol starch syrup, oligosaccharides, aspartame, sucralose, acesulfame potassium, saccharin, saccharin sodium, stevia, stevia extract, stevioside, neotame, alitame, thaumatin, neohesperidin dihydrochalcone, paramethoxycinnamic aldehyde, perillartine, and licorice. Acidulants include acetic acid, lactic acid, citric acid, etc. Examples of bulking agents include sugars, polysaccharides, modified starch, casein, gelatin, carboxymethylcellulose, and lecithin. Examples of pigments include natural pigments and organic synthetic pigments. Specific examples include hibiscus pigment, huckleberry pigment, plum pigment, seaweed pigment, dewberry pigment, grape juice pigment, blackberry pigment, blueberry pigment, mulberry pigment, morello cherry pigment, red currant pigment, loganberry pigment, paprika powder, malt extract, rutin, flavonoids, red cabbage pigment, red radish pigment, adzuki bean pigment, turmeric pigment, olive tea, cowberry pigment, chlorella powder, saffron pigment, perilla pigment, strawberry pigment, chicory pigment, pecan nut pigment, monascus pigment, safflower pigment, purple sweet potato pigment, lac pigment, spirulina pigment, onion pigment, tamarind pigment, chili pepper pigment, gardenia pigment, caramel pigment, lithospermum root pigment, rosewood pigment, krill pigment, orange pigment, carrot carotene, Blue No. 1, Yellow No. 4, and Green No. 3. Examples of emulsifiers include fatty acid monoglycerides, fatty acid diglycerides, fatty acid triglycerides, propylene glycol fatty acid esters, sucrose fatty acid esters, polyglycerol fatty acid esters, lecithin, enzyme-treated lecithin, starch, modified starch, dextrin, sorbitan fatty acid esters, quillaja extract, gum arabic, tragacanth gum, guar gum, karaya gum, xanthan gum, pectin, alginic acid and salts thereof, carrageenan, gelatin, and casein. Functional substances refer to substances that have nutritional or biological regulation functions, and examples thereof include docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), DHA and / or EPA-containing fish oil, animal and vegetable oils and fats such as linoleic acid, γ-linolenic acid, α-linolenic acid, lecithin, diacylglycerol, and derivatives thereof, animal and vegetable extracts such as rosemary, sage, perilla oil, chitin, chitosan, royal jelly, and propolis, vitamin A, vitamin D, vitamin E, vitamin F, vitamin K, Vitamins such as coenzyme Q10 and alpha lipoic acid, coenzymes and their derivatives, polyphenols such as gamma-oryzanol, catechin, anthocyanin, isoflavone, rutin, chlorogenic acid, and theaflavin, dietary fiber such as indigestible dextrin, carbohydrates such as palatinose, xylitol, and oligosaccharides, salts such as calcium citrate malate (CCM), substances derived from milk proteins such as casein phosphopeptides, lactoferrin, and milk peptides, heme iron, sodium fluoride, and fluoride. Examples of effective anti-inflammatory agents include fluorides such as potassium fluoride, stannous fluoride, strontium fluoride, and sodium monofluorophosphate; water-soluble phosphate compounds such as potassium and sodium orthophosphate; tranexamic acid, epsilon aminocaproic acid, dl-tocopherol acetate, α-bisabolol, dihydrocholesterol, chlorhexidine salts, azulene, glycyrrhetin, glycyrrhetinic acid, glycyrrhizinic acid and its salts, copper chlorophyllin sodium, chlorophyll, and chelating phosphate compounds such as glycerophosphate; copper compounds such as copper gluconate; aluminum lactate, strontium chloride, potassium nitrate, hydroxamic acid and its derivatives, sodium tripolyphosphate, methoxyethylene, epidihydrocholesterol, aluminum allantoin chlorohydroxy, ascorbic acid, lysozyme chloride, isopropylmethylphenol, benzethonium chloride, cetylpyridinium chloride, trichlorocarbanilide, zinc citrate, and Phellodendron Bark extract. Milk components include raw milk, cow's milk, whole milk powder, skim milk powder, and cream, as well as milk proteins such as casein and whey, and those derived from milk of goat or sheep, or their decomposition products. Examples of flavor improvers include sucralose, cyclodextrin, theanine, hesperidin glycoside, and sugarcane extract. Examples of the various flavor materials that can be used include natural fragrances, natural essential oils, and various synthetic fragrances. These fragrances are not particularly limited as long as they can be used in foods and beverages, and examples include esters, aldehydes, (thio)ethers, alcohols, ketones, lactones, carboxylic acids, aliphatic hydrocarbons, nitrogen-containing heterocyclic compounds, sulfur-containing heterocyclic compounds, amines, thiols, phenols, and essential oils. Specific compounds include acetaldehyde, ethyl acetoacetate, acetophenone, anethole, anisaldehyde, amyl alcohol, α-amylcinnamaldehyde, allyl cyclohexane propionate, methyl anthranilate, ambrettolide, ionone, isoamyl alcohol, isoeugenol, isoamyl isovalerate, ethyl isovalerate, isothiocyanates, allyl isothiocyanate, isovaleraldehyde, isobutanol, isobutyraldehyde, isopropanol, isopentylamine, indole and its derivatives, γ-undecalactone, ethyl acetate, a mixture of 2-ethyl-3,5-dimethylpyrazine and 2-ethyl-3,6-dimethylpyrazine, ethyl thioacetate, ethyl vanillin, 2-ethylpyrazine, ethyl butyrate, 2-ethyl-3-methylpyrazine, and 2-ethyl-5-methylpyrazine. , 5-ethyl-2-methylpyrazine, ethyl methylphenylglycidate, ethyl lactate, eugenol, octanal, ethyl octanoate, capsaicin, carbyl acetate, carvone, isoamyl formate, geranyl formate, citronellyl formate, cinnamic acid, ethyl cinnamate, methyl cinnamate, ketones, geraniol, isoamyl acetate, ethyl acetate, geranyl acetate, cyclohexyl acetate, citronellyl acetate, cinnamyl acetate, terpinyl acetate, phenethyl acetate, butyl acetate, benzyl acetate, l-menthyl acetate, linalyl acetate, ethyl salicylate, methyl salicylate, 2,3-diethyl-5-methylpyrazine, allyl cyclohexylpropionate, citral, citronellal, citronellol, 1,8-cineole, dimethyl sulfide, 2,3-dimethylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2,6-Dimethylpyridine, gingerol, cinnamyl alcohol, cinnamaldehyde, spilanthol, thymol, decanal, decanol, ethyl decanoate, 5,6,7,8-tetrahydroquinoxaline, 2,3,5,6-tetramethylpyrazine, terpineol, 2,3,5-trimethylpyrazine, γ-nonalactone, vanillyl butyl ether, vanillin, paramethylacetophenone, valeraldehyde, hydroxycitronellal, hydroxycitronellal dimethyl acetal, pinene, piperidine, piperine, piperonal pyrazine, pyrrolidine, isoamyl phenylacetate, isobutyl phenylacetate, ethyl phenylacetate, 2-(3-phenylpropyl)pyridine, phenethyl Aminomethyl methyl amine, phenoxyethyl isobutyrate, fenchone, butanol, butylamine, butyraldehyde, furfural and its derivatives, pulegone, propanol, propionaldehyde, propionic acid, isoamyl propionate, ethyl propionate, benzyl propionate, hexanoic acid, allyl hexanoate, ethyl hexanoate, hexanal, hexenol, ethyl heptanoate, perillaldehyde, benzyl alcohol, benzaldehyde, 2-pentanol, 1-penten-3-ol, d-borneol, maltol, methyl anthranilate, methyl N-methylanthranilate, methyl epijasmonate, 5-methylquinoxaline, 6-methylquinoline, 5-methyl-6,Examples of the menthol isomers include 7-dihydro-5H-cyclopentapyrazine, methyl β-naphthyl ketone, 2-methylpyrazine, 2-methylbutanol, 3-methyl-2-butanol, 2-methylbutyraldehyde, 3-methyl-2-butenal, 3-methyl-2-butenol, menthyl acetate, and 1-menthol, as well as menthone, butyric acid, isoamyl butyrate, ethyl butyrate, cyclohexyl butyrate, butyl butyrate, gamma and delta lactones having 4 to 12 carbon atoms, linalyl acetate, linalool, and limonene. Specific essential oils include anise oil, anise star oil, bergamot oil, basil oil, West Indian bay leaf oil, galbanum oil, apple oil, apricot oil, cassia oil, camphor oil, buchu leaf oil, cardamom seed oil, cassia bark oil, Roman spider lily oil, cinnamon bark oil, cinnamon leaf oil, clove bud oil, konjac green oil, coriander oil, cubeba oil, caraway oil, sweet fennel oil, garlic oil, ginger oil, petitgrain oil, lemon oil, lime oil, orange oil, citrus oil, cedar oil, citronella oil, patchouli oil, eucalyptus oil, bay oil, grapefruit oil, mandarin oil, sandalwood oil, juniper nut oil, rose oil, and bayberry oil. Examples of such oils include orchid oil, tangerine oil, geranium oil, wintergreen oil, clove oil, thyme oil, sage oil, cardamom oil, coriander oil, lime oil, yuzu oil, lavender oil, rosemary oil, laurel oil, perilla oil, chamomile oil, caraway oil, marjoram oil, celery oil, bay oil, origanum oil, pine needle oil, neroli oil, jasmine oil, patchouli oil, paracles oil, orris concrete, rose absolute, orange flower absolute, vanilla absolute, patchouli absolute, and processed products thereof (such as front distillation, back distillation, fractional distillation, liquid-liquid extraction, essence preparation, and powdered fragrance preparation).

[0012] The polyphenol concentration in the food material of the present invention may be 0.76 mmol / g or less, 0.42 mmol / g or less, or 0.17 mmol / g or less, or 0.001 mmol / g or more, 0.01 mmol / g or more, 0.02 mmol / g or more, or 0.17 mmol / g or more, preferably 0.17 mmol / g or more, calculated as catechol. In the present invention, the polyphenol concentration in catechol terms refers to the polyphenol concentration in catechol terms calculated from a calibration curve (relationship between catechol molecule concentration and absorbance in a standard substance) using polyphenols having one or more catechol structures in the molecule as standard substances according to the Folin-Ciocalteu method (Am. J. Enol. Vitic., 16, 144 (1965)). The polyphenol oxidase concentration in the food material of the present invention may be 1,000 units / g or less, 100 units / g or less, 80 units / g or less, 40 units / g or less, or 25 units / g or less, or 1 unit / g or more, 5 units / g or more, 25 units / g or more, or 40 units / g or more, preferably 25 units / g or more. The unit of enzyme activity is defined as the enzyme activity that increases the ultraviolet absorbance at 420 nm by 1 unit when 1.7 mg / mL of chlorogenic acid as a substrate is reacted in 5 mL of phosphate buffer (pH 6.5) at 25°C for 5 minutes, 1 mL of 10% sulfuric acid is added to terminate the enzymatic reaction, and insoluble matter is removed using a PVDF membrane filter with a pore size of 0.45 μm. Furthermore, the enzyme activity per 1 mmol of polyphenols in the food material of the present invention may be 0.1 units or more, 1 unit or more, 12 units or more, 36 units or more, 60 units or more, or 235 units or more, but it is preferable to contain polyphenol oxidase in an amount that results in 10 units or more, and it is more preferable to contain polyphenol oxidase in an amount that results in 60 units or more. The food material of the present invention preferably has an activity of 5,000 or more, and more preferably 14,000 or more, per unit mass (g) of the agent due to polyphenols and polyphenol oxidase. Here, the activity of polyphenols and polyphenol oxidase refers to the binding activity of polyphenol quinones resulting from the reaction of polyphenols with polyphenol oxidase with methyl mercaptan, and is evaluated by the following method: 20 mg of a test substance (i.e., a food material) containing an active ingredient is added to 2 ml of 0.015% methyl mercaptan sodium / 50 mM phosphate buffer (pH 6.5) placed in a 50 ml cylindrical bottle (inner diameter 3.3 cm, height 6 cm), and the bottle is sealed and incubated at 25°C for 20 minutes. The decrease in the concentration of volatile methyl mercaptan in the headspace of the system due to the active ingredient is measured. The activity amount in the present invention is defined as the activity that reduces the volatile methyl mercaptan in the head space of the system by 1 ppm in the evaluation system for methyl mercaptan binding activity. The mass ratio of polyphenols to polyphenol oxidase in the food material of the present invention can be adjusted appropriately to optimize the activity of the polyphenols and polyphenol oxidase. For example, the mass ratio of polyphenols (contents) to polyphenol oxidase (contents) may be 1:99, 10:90, 20:80, 50:50, 80:20, 90:10, or 99:1. For example, the mass ratio of polyphenol-containing green coffee bean extract to polyphenol oxidase-containing burdock powder may be 1:99, 10:90, 20:80, 50:50, 80:20, 90:10, or 99:1, with a range of 20:80 to 50:50 being more preferred. When used to regulate oral immunity, the intake amount of the food material of the present invention can be appropriately set so that an activity level of 15 or more, preferably 25 or more, and more preferably 50 or more can be ingested per serving. When used to regulate gastrointestinal or systemic immunity, the intake amount can be appropriately set so that an activity level of 1000 or more, 2500 or more, and preferably 5000 or more can be ingested per serving. [Example]

[0013] 1. Preparation of each sample 1-1 Preparation of green coffee bean extract (polyphenol-containing plant extract) After grinding the green coffee beans in a grinder, the beans that did not pass through a 5mm mesh were removed, and water was added and the beans were extracted for 2 hours at 85-95°C. The extract was filtered, and the filtrate was adsorbed onto an XAD-2 (Organo Corporation) column. After washing with water, the eluate was eluted with methanol and concentrated to dryness to obtain the green coffee bean extract. The polyphenol content of the resulting green coffee bean extract was 0.84 mmol / g. The polyphenol content was measured by the Folin-Ciocalteu method (Am. J. Enol. Vitic., 16, 144 (1965)). The polyphenol concentration of the extract was expressed as a catechol equivalent calculated from a calibration curve (relationship between catechol molecule concentration and absorbance) using chlorogenic acid as the standard substance. 1-2 Preparation of burdock powder (polyphenol oxidase) The washed burdock and -20°C acetone were placed in a mixer, ground, and then suction filtered. The residue was thoroughly washed with 80% aqueous acetone at 5°C, combined with the filtrate, and the acetone was distilled off. The mixture was then freeze-dried to obtain burdock powder. The enzyme specific activity of the resulting burdock powder was 50 units / g. The specific enzyme activity of the resulting burdock powder was determined by incubating 20 mg of burdock powder in 5 ml of phosphate buffer (pH 6.5) with 1.7 mg / mL chlorogenic acid (Tokyo Kasei) as the substrate at 25°C for 5 minutes, followed by the addition of 1 ml of 10% sulfuric acid to terminate the enzymatic reaction. After removing insoluble matter using a 0.45 μm pore size PVDF membrane filter, the solution was measured for UV absorption at 420 nm (absorbance (substrate + enzyme)). Similar experiments were performed without enzyme and without substrate, and the absorbance was measured under each condition (absorbance (substrate only) and absorbance (enzyme only) respectively). The enzyme and substrate-free conditions served as controls, and the enzyme activity required to increase the absorbance at 420 nm by 1 unit in 5 minutes under the enzyme and substrate conditions was defined as 1 unit. Specifically, enzyme activity was calculated using the following formula: Enzyme activity (Unit / g) = {(absorbance (substrate + enzyme) / enzyme weight (mg) - (absorbance (enzyme only) / enzyme weight (mg) - absorbance (substrate only)} x {1000 / enzyme weight (mg))

[0014] 2. Study of inflammatory cytokine regulation Samples A to D were prepared as shown in Table 1, and tests 1 to 4 were carried out.

[0015] [Table 1]

[0016] [Table 2]

[0017] 2-1 Test 1 (Data showing that PP+PPO composition inhibits IL-6 expression in THP-1 cells) In the following test, the anti-inflammatory effect of a PP+PPO composition (Sample A) consisting of green coffee bean extract and burdock powder (1:1 (mass ratio)) on human monocyte-type cells THP-1 (obtained from ATCC) was evaluated. Sample A was suspended in distilled water at a concentration of 10 mg / ml, and then centrifuged to remove water-insoluble matter. 6 The human monocytic cell line THP-1 was cultured at 10% of the culture medium. After 20 hours of culture, lipopolysaccharide (LPS) (#3024, WAKO) was added at a concentration of 10 ng / ml, and the cells were harvested 4 hours later. The mRNA expression of the inflammatory cytokine IL-6 in the cells was measured by qPCR. The results are shown in Table 3. The IL-6 expression levels in the table are relative to the IL-6 expression level upon LPS stimulation, which is set to 1. The mRNA expression level analysis was carried out as follows. Total RNA was extracted from the harvested cells using the ReliaPrep RNA Cell Miniprep System (#Z6012, Promega) or RNAzol (#RN190, Cosmo Bio). cDNA was synthesized by RT using the RevaTraAce qPCR RT Master Mix (#FSQ-201, TOYOBO). Quantitative PCR was performed using the StepOne Real-Time PCR System (Applied Biosystems) and THUNDERBIRD SYBR qPCR Mix (#QPS-101, TOYOBO).

[0018] [Table 3]

[0019] 2-2 Test 2 (Data showing that the PP+PPO composition has a superior inhibitory effect on IL-6 expression in THP-1 cells compared to PP alone or PPO alone) In the following tests, the anti-inflammatory effects of a PP+PPO composition (Sample A) consisting of green coffee bean extract and burdock powder (1:1 (mass ratio)), a PP+PPO composition (Sample B) consisting of green coffee bean extract and burdock powder (2:8 (mass ratio)), a PP composition (Sample C) consisting of green coffee bean extract, and a PPO composition (Sample D) consisting of burdock powder on human monocyte-type cells THP-1 were evaluated. The evaluation method was the same as in Test 1 above. The results are shown in Table 4.

[0020] [Table 4]

[0021] 2-3 Test 3 (Data showing that the PP+PPO composition inhibits the expression of IL-6, TNF, and IL-1β in PBMC cells) In the following study, the same test as in Study 1 was performed on human-derived peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from the peripheral blood of healthy individuals by sucrose density gradient analysis using lymphocyte separation medium 1077 (obtained from multiple manufacturers). In addition to the inflammatory cytokine IL-6, the expression levels of TNF and IL-1β were also evaluated. The results are shown in Table 5.

[0022] [Table 5]

[0023] 2-4 Test 4 (Data demonstrating that the PP+PPO composition has a superior inhibitory effect on IL-6 expression in PBMC cells compared to PP or PPO alone) In the following test, a test similar to Test 2 was performed on human-derived monocytic PBMC cells. The results are shown in Table 6.

[0024] [Table 6]

Claims

1. A food material containing polyphenols and polyphenol oxidase as active ingredients, which has excellent immune-regulating effects by adjusting the amount of cytokines in cells.

2. 2. The food material according to claim 1, wherein the polyphenols include o-diphenol polyphenols and / or p-diphenol polyphenols.

3. 2. The food material according to claim 1, wherein the polyphenol comprises a polyphenol derived from a polyphenol-containing plant or an extract thereof.

4. 2. The food material according to claim 1, wherein the polyphenol oxidase comprises a polyphenol oxidase derived from a polyphenol oxidase-containing plant or an extract thereof.

5. The food material according to claim 1 , wherein regulating the amount of cytokines comprises negatively regulating an increase in the amount of cellular inflammatory cytokines induced by stimulation with pathogen-associated molecular patterns (PAMPs).

6. The food material according to claim 5, which regulates the amount of inflammatory cytokines in cells when the pathogen-associated molecular pattern (PAMP) is LPS.

7. 7. The food material according to claim 6, wherein the inflammatory cytokine is selected from the group consisting of IL-6, TNF-α, and IL-1β.

8. The food material according to claim 1 , wherein the cells comprise monocyte cells.

9. An immune-modulating food or drink containing the food material according to any one of claims 1 to 8.

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