Heat stress mitigating agent and heat stress mitigation method

Abstract

The present invention provides a heat stress mitigating agent and a method for mitigating heat stress. [Solution] A heat stress reliever containing olive extract as an active ingredient. According to the present invention, the suppression of cell proliferation and melanin production caused by heat stress are alleviated.

Description

Technical Field

[0001] The present invention relates to a heat stress reliever and a heat stress relief method. More specifically, it relates to a heat stress inhibitor and a heat stress inhibition method using olive extract.

Background Art

[0002] In recent years, due to the influence of global warming, the temperature has risen, and problems such as livestock suffering from heat stress and developing heat stroke or dying from heat have occurred. As methods for alleviating these heat stresses, for example, methods of administering glycosyl hesperetin and adrenal medulla hormones have been reported (Patent Document 1 and Patent Document 2), but the effects are not sufficient.

[0003] In addition, melanin is produced by ultraviolet rays, which causes freckles, chloasma, and pigmentation after sunburn, becoming a problem for the skin. In order to overcome these problems, sunscreen agents and melanin production inhibitors that prevent ultraviolet rays have been actively developed (for example, Patent Document 3). However, it is known that not only ultraviolet rays but also heat stress can damage cells and cause melanin production (Non-Patent Document 1). It is also known that near-infrared rays can cause an increase in skin temperature, which can also cause heat stress. Near-infrared rays penetrate deeper into the skin than ultraviolet rays and are considered to cause heat stress.

[0004] Skin external preparations having a near-infrared ray protection effect have also been developed (for example, Patent Document 4), but there is a problem that this protects near-infrared rays on the surface of the skin and does not relieve heat stress from the inside of the body. Therefore, there has been a demand for the development of drugs with a high heat stress relief effect and heat stress relief methods.

Prior Art Documents

Non-Patent Documents

[0005]

Non-Patent Document 1

[0006] [Patent Document 1] WO2020 / 138026 [Patent Document 2] Japanese Patent Publication No. 2008-13521 [Patent Document 3] Japanese Patent Application Publication No. 10-265325 [Patent Document 4] Japanese Patent Publication No. 2022-176168 [Overview of the project] [Problems that the invention aims to solve]

[0007] The present invention provides a heat stress mitigating agent and a method for mitigating heat stress. [Means for solving the problem]

[0008] (1) A heat stress reliever containing olive extract as an active ingredient. (2) Heat stress is a decrease in cell count, and a heat stress mitigator as described in (1). (3) Heat stress is caused by an increase in melanin production, and the heat stress mitigator described in (1). (4) A heat stress mitigation composition comprising any of the agents described in (1) to (3). (5) A heat stress relief composition, wherein the composition is a cosmetic, quasi-drug, pharmaceutical, or food / beverage. (6) A method for alleviating heat stress, comprising the step of administering olive extract. (7) Heat stress is a decrease in cell count, and (6) methods for mitigating heat stress. (8) Heat stress is caused by an increase in melanin production, and (6) methods for alleviating heat stress. The preferred method involves applying the treatment to cultured cells (including animal cultured cells) or animals.

Advantages of the Invention

[0009] According to the present invention, there are provided a composition and a method for alleviating the suppression of cell growth and the production of melanin caused by heat stress.

Brief Description of the Drawings

[0010] [Figure 1] FIG. 1 is a graph showing the suppression of the decrease in the number of cells induced by heat stress with olive extract. [Figure 2] FIG. 2 is a graph showing the decrease in the number of cells caused by heat stress. [Figure 3] FIG. 3 is a graph showing the increase in the amount of melanin caused by heat stress. [Figure 4] FIG. 4 is a graph showing the suppression of the decrease in the number of cells induced by heat stress with olive extract. [Figure 5] FIG. 5 is a diagram showing the suppression of the increase in the amount of melanin induced by heat stress with olive extract.

Modes for Carrying Out the Invention

[0011] In the process of screening a drug capable of alleviating heat stress, the inventors found that olive extract has an activity of alleviating heat stress, and completed the present invention. That is, the present invention is a heat stress reliever containing olive extract as an active ingredient.

[0012] In the present specification, "heat stress" means culturing at a temperature higher than normal during cell culture. Heat stress causes a decrease in the number of cell proliferations and / or an increase in the accumulation amount of melanin, but may also cause other effects. Heat stress at the individual level of an animal means that the body temperature cannot be controlled and the body temperature rises. Heat stroke may occur due to heat stress. Since heat stroke can also be caused by infrared radiation reflection and the like, the composition of the present invention is also a heat stress reliever for infrared radiation. <00,00090> "Alleviation of heat stress" means that the effects of the above heat stress are weakened. "Suppression of heat stress" has the same meaning. Due to heat stress, in cultured cells, cell growth disorders and functional disorders such as a decrease in cell number, an increase in the amount of melanin, and in animals such as livestock, a decrease in reproductive ability due to heat stroke are known.

[0014] Olive (scientific name: Olea europaea) belongs to the genus Olea of the family Oleaceae, is a dicotyledonous plant, an evergreen hardwood tree, and is a tall tree. The leaves are oblong and hard, and the back surface is silver-white and densely covered with short hairs.

[0015] The preparation of olive extract (olive extract) was carried out by water extraction by adding distilled water to dried olive leaves. The extraction is carried out at room temperature, and the extraction time is preferably 12 hours or more, more preferably 15 hours or more, still more preferably 18 hours or more, particularly preferably 24 hours or more, and the upper limit of the extraction time is 36 hours or less, more preferably 30 hours or less. The extraction site is most preferably the leaves, but fruits, flowers, stems, and roots may also be used.

[0016] The extraction solvent for olive extraction is not limited to water, and water-soluble organic solvents or water-containing organic solvents such as BG (1,3-butylene glycol) or BG + water may also be used. The water content of the water-containing organic solvent is preferably 20 to 70% (v / v) (that is, the 1,3-butylene glycol is 30 to 80%). When extracted with a water-containing organic solvent, after extraction, the organic solvent may be distilled off using a rotary evaporator or the like. If necessary, it may be adsorbed onto a synthetic adsorption resin (for example, Diaion HP resin (manufactured by Mitsubishi Chemical Corporation), Amberlite XAD resin (manufactured by Rohm and Haas Company), Duolite S resin (manufactured by Diamond Shamrock Corporation), etc.). After adsorption, impurities are washed with water or water added with a low-content organic solvent, eluted from the synthetic resin using an organic solvent or a mixture of an organic solvent and water, concentrated, and dried to obtain an extract solid content. Examples of the drying method include, but are not limited to, spray drying and evaporation to dryness.

[0017] The obtained extract was filtered to obtain olive extract. It was found that adding this extract to cultured fibroblasts suppressed the reduction in cell number due to heat stress (Figure 1). When subjected to heat stress (42°C, 3 hours), the cell number decreased by about 10% in the control group, but in the group to which olive extract was added, the cell number recovered to almost 100%.

[0018] Similar results were obtained when using melanocytes (melanin-forming cells) (Figures 2 and 4). This indicates that the olive extract of the present invention can suppress the decrease in cell number in cell culture due to heat stress.

[0019] Next, the olive extract was added to cultured melanocytes to investigate whether melanin production was suppressed. In the control group, melanin production increased by approximately 40% under heat stress (Figure 3). In contrast, melanocytes treated with olive extract showed a decrease of more than 20% in melanin production compared to the control group (Figure 5). These data indicate that olive extract can suppress melanin accumulation caused by heat stress.

[0020] Because olive oil has been used as food for centuries, olive extract is considered highly safe. Therefore, pharmaceuticals, quasi-drugs, cosmetics, food and beverages, and animal feed containing olive extract also have the advantage of being highly safe and having few side effects.

[0021] When using olive extract with a solid content of 1% by weight, the preferred upper limit of addition to the culture medium is preferably 1 / 1000 (0.001% by weight) or less, 1 / 3000 or less, 1 / 5000 or less, 1 / 8000 or less, and 1 / 10000 or less, based on weight / weight. The lower limit of addition is 1 / 50000 or more, 1 / 30000 or more, 1 / 20000 or more, and 1 / 10000 or more. Any combination of these above and below values ​​is acceptable (excluding ranges that are theoretically impossible). This also applies to the concentration ranges described below.

[0022] The heat stress mitigating agent (mitigating composition) of the present invention has been shown to be effective in cultured cells, but is also considered applicable to humans. When applied to humans, the composition of the present invention can be taken as is or dissolved in water and taken orally, but it may also be formulated by adding various excipients. Examples of formulations include granules, tablets, and capsules. In the case of cultured cells, the amount to be added is preferably 0.01% to 50% by weight, and more preferably the upper limit of the amount to be added is 40% or less by weight, 30% or less by weight, 20% or less by weight, 10% or less by weight, 5% or less by weight, 3% or less by weight, and 1% or less by weight.

[0023] The heat stress reliever of the present invention can be used as is, but it can also be mixed with appropriate additives to form a composition, provided that the effects of the present invention are not impaired. Examples of compositional forms include cosmetics, pharmaceuticals, quasi-drugs, food and beverages, and animal feed. For example, when the agent of the present invention is used to remove accumulated melanin and for skin whitening purposes, it can be in the form of a topical skin composition such as a cosmetic. Furthermore, when the agent of the present invention is used for the treatment, improvement, and prevention of pigmentation (spots), it is preferable to use it in the form of a pharmaceutical.

[0024] When the agent of the present invention is incorporated into cosmetics or quasi-drugs, the dosage form may be any of the following: aqueous solution, solubilized, emulsified, powder, powder dispersion, oil-liquid, gel, ointment, aerosol, water-oil two-layer system, or water-oil-powder three-layer system. Furthermore, the cosmetics or quasi-drugs can be manufactured by selecting and appropriately incorporating various components, additives, bases, etc., commonly used in topical skin compositions, along with the olive extract mentioned above, according to the methods known in the art. The form may be any of the following: liquid, emulsion, cream, gel, paste, spray, etc. The ingredients include, for example, oils and fats (olive oil, coconut oil, evening primrose oil, jojoba oil, castor oil, hydrogenated castor oil, etc.), waxes (lanolin, beeswax, carnauba wax, etc.), hydrocarbons (liquid paraffin, squalene, squalane, petrolatum, etc.), fatty acids (lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, etc.), higher alcohols (myristyl alcohol, cetanol, cetostearyl alcohol, stearyl alcohol, behenyl alcohol, etc.), and esters (isopropyl myristate, i) Examples of ingredients include sopropyl, cetyl octanoate, glyceryl trioctanoate, octyldodecyl myristate, octyl stearate, stearyl stearate, etc.), organic acids (citric acid, lactic acid, α-hydroxyacetic acid, pyrrolidone carboxylic acid, etc.), sugars (maltitol, sorbitol, xylobiose, N-acetyl-D-glucosamine, etc.), proteins and protein hydrolysates, amino acids and their salts, vitamins, plant and animal extracts, various surfactants, humectants, UV absorbers, antioxidants, stabilizers, preservatives, disinfectants, fragrances, etc.

[0025] Examples of cosmetics and quasi-drugs include lotions, emulsions, gels, serums, general creams, sunscreens, packs, masks, facial cleansers, cosmetic soaps, foundations, face powders, and body lotions.

[0026] When the above olive extract is incorporated into a pharmaceutical product, it can be mixed with pharmacologically and pharmaceutically acceptable additives and formulated into various formulations suitable for application to the affected area. Pharmacologically and pharmaceutically acceptable additives may include, depending on the dosage form and use, appropriately selected formulation bases or carriers, excipients, diluents, binders, lubricants, coatings, disintegrants or disintegration aids, stabilizers, preservatives, antiseptics, bulking agents, dispersants, wetting agents, buffers, solubilizers or solubilizers, isotonic agents, pH adjusters, propellants, colorants, sweeteners, flavoring agents, fragrances, etc., which can then be prepared into various formulations that can be administered orally or parenterally systemically or topically by various known methods. When providing the pharmaceutical product of the present invention in any of the above forms, it can be manufactured by methods commonly used by those skilled in the art, such as those indicated in the individual articles of the General Provisions for Formulations of the Japanese Pharmacopoeia [2].

[0027] When formulating the composition, additives can be incorporated to the extent that they do not impede the purpose of the composition of the present invention. Examples of additives include dietary fibers and thickeners derived from sources other than vegetables, such as hemicellulose, lignin, guar gum, konjac mannan, isagol, alginic acid, agar, carrageenan, chitin, carboxymethylcellulose, and polydextrose; minerals such as calcium, iron, sodium, zinc, copper, potassium, phosphorus, magnesium, iodine, manganese, and selenium; fat-soluble or water-soluble vitamins such as vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, niacin, folic acid, and pantothenic acid; glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, phospholipids, gum arabic, and xanthamine. Examples of emulsifiers and dispersants such as tung gum, tragacanth gum, and locust bean gum, as well as fillers, excipients, lubricants, preservatives / antioxidants, flavor modifiers, fragrances, formulation bases and carriers, diluents, binders, coatings, disintegrants or disintegration aids, stabilizers, preservatives, fillers, dispersants, wetting agents, buffers, solvents or solubilizers, isotonic agents, pH adjusters, propellants, colorants, sweeteners, flavoring agents, flavorings such as sodium chloride, monosodium glutamate, glycine, succinic acid, and sodium lactate, acidulants such as citric acid, sodium citrate, acetic acid, adipic acid, fumaric acid, and malic acid, low-calorie sweeteners such as maltitol and aspartame, and colorants, but not limited to these.

[0028] [Optional ingredients] The heat stress mitigation composition of the present invention may optionally contain other components besides olive extract. Examples of other components include physiologically active components other than olive extract. Other examples include disintegrants, sugar alcohols, excipients, fluidizers, lubricants, binders, flavoring agents, colorants, surfactants, adsorbents, antistatic agents, disintegration extenders, film-forming components, and plasticizers, which can be appropriately selected and used mainly depending on the dosage form. Hereafter, examples of components other than physiologically active components will be explained using tablets (functional foods such as supplements) as an example.

[0029] -Other bioactive ingredients- Examples of physiologically active ingredients other than olive extract include plant-derived components other than olive, vitamins (fat-soluble and water-soluble), minerals, dietary fiber, fish or algae-derived components, lactic acid bacteria (live and dead cells), proteins, peptides, and carbohydrates.

[0030] Other plant-derived ingredients besides olive include, for example, cocoa extract, long pepper extract, kudzu flower extract, African mango extract, Darvilleia extract, ginkgo leaf extract, grape seed extract, pine bark extract, Coleus forskohlii extract, artichoke leaf extract, rosemary extract, rooibos extract, tomato extract, Panax notoginseng extract, Panax notoginseng acid-treated product, licorice extract, Salacia extract, passion fruit seed extract, mangosteen extract, and other plant extracts, as well as processed products and purified compounds thereof. Examples of purified compounds include raspberry ketone, isoflavone, glucosylceramide, hesperidin, monoglucosylhesperidin, β-carotene, GABA, piceatannol, glucosylceramide, panaxadiol, and panaxatriol.

[0031] Examples of vitamins (fat-soluble and water-soluble), minerals, and dietary fiber include vitamin C, vitamin A, vitamin E, vitamin K, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin D, niacin, pantothenic acid, folic acid, biotin, potassium, calcium, iron, zinc, sodium, magnesium, phosphorus, selenium, copper, chromium, manganese, iodine, indigestible dextrin, isomaltodextrin, dextrin, inulin, psyllium husk, and guar gum hydrolysate. These vitamins, minerals, and dietary fiber may be derived from natural sources such as plants, or they may be artificially synthesized.

[0032] Examples of components derived from fish or algae include astaxanthin, docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), anserine, carnosine, fucoidan, and phycocyanin. Examples of carbohydrates include indigestible oligosaccharides.

[0033] Examples of lactic acid bacteria include those belonging to genera such as Lactobacillus, Lacticaseibacillus, Lactipruntilassirus, Lactilactobacillus, Ligylactobacillus, Leviractobacillus, Limosilactobacillus, Enterococcus, Lactococcus, Streptococcus, Staphylococcus, and Bifidobacterium.

[0034] Examples of lactic acid bacteria belonging to the genus Lacticaseibacillus include Lacticaseibacillus casei, Lacticaseibacillus paracasei, and Lacticaseibacillus rhamnosus. Examples of lactic acid bacteria belonging to the genus Lactobacillus include Lactobacillus gasseri and Lactobacillus paragasseri. Examples of lactic acid bacteria belonging to the genus Lactococcus include Lactococcus lactis and Lactococcus cremoris. Examples of lactic acid bacteria belonging to the genus Leviractobacillus include Leviractobacillus brevis. Examples of Enterococcus lactic acid bacteria include Enterococcus faecalis and Enterococcus faecium.

[0035] Examples of proteins include lactoferrin, and examples of peptides include collagen peptides, elastin peptides, and placenta-derived peptides.

[0036] It is preferable that these bioactive ingredients include at least one selected from rooibos extract, passion fruit seed extract, mangosteen extract, tomato extract, GABA, piceatannol, glucosylceramide, vitamins, astaxanthin, lactic acid bacteria (Lacticaseibacillus and Lactococcus), collagen peptides, elastin peptides, placenta-derived peptides, and lactoferrin. When these are included in combination with olive extract, it is expected that the skin-improving effect will be further enhanced.

[0037] -Disintegrant- Examples of disintegrants include cellulose derivatives, starch, and its derivatives.

[0038] Examples of the cellulose derivatives include hydroxypropylcellulose, carboxymethylcellulose (carmellose), sodium carboxymethylcellulose (carmellose sodium, CMC-Na), calcium carboxymethylcellulose (also known as carmellose calcium, CMC-Ca), croscarmellose sodium, cross-linked insoluble polyvinylpyrrolidone, and crospovidone. Of these, calcium carboxymethylcellulose, carboxymethylcellulose, and sodium carboxymethylcellulose are preferred from the viewpoint of improving disintegrability.

[0039] The starch may be of natural origin or artificially produced, and examples include wheat starch, rice starch, corn starch, potato starch, and tapioca starch. Examples of starch derivatives include partially pregelatinized starch, pregelatinized starch, acetylated phosphate cross-linked starch, acetylated oxidized starch, acetylated adipic acid cross-linked starch, sodium octenyl succinate starch, starch acetate, oxidized starch, sodium carboxymethyl starch (CM-starch), hydroxypropylated phosphate cross-linked starch, hydroxypropylated starch, phosphorylated starch, phosphate cross-linked starch, and phosphate monoesterified phosphate cross-linked starch.

[0040] Of the starches and their derivatives, those derived from maize are preferred from the viewpoint of improving disintegrability. Specifically, preferred maize starches and their derivatives include maize starch, partially pregelatinized starch, pregelatinized starch, acetylated starch, hydroxypropylated starch, phosphorylated starch, phosphate-crosslinked starch, oxidized starch, octenyl succinate oxidized starch, acetylated phosphate-crosslinked starch, acetylated adipic acid-crosslinked starch, hydroxypropylated phosphate-crosslinked starch, and phosphate-monoesterified phosphate-crosslinked starch. More preferred are maize starch, partially pregelatinized starch, and pregelatinized starch, with partial pregelatinized starch being even more preferred.

[0041] The disintegrant preferably contains one of a cellulose derivative and / or starch or its derivatives, more preferably at least two, and even more preferably all of them. Each of the cellulose derivative and / or starch or its derivatives may be a single type or a combination of two or more types.

[0042] When a cellulose derivative is included, its content is usually 1% by mass or more relative to the total mass of the food composition. This improves disintegration and can accelerate the disintegration time. The upper limit is usually 3% by mass or less, preferably 2.5% by mass or less, and more preferably 2% by mass or less. This can improve moldability. Therefore, the content of the cellulose derivative is usually 1 to 3% by mass, preferably 1 to 2.5% by mass, and more preferably 1 to 2% by mass.

[0043] When starch or its derivatives are included, their content is usually 3% by mass or more, preferably 7% by mass or more, and more preferably 8% by mass or more, relative to the total mass of the food composition. This improves disintegration and can shorten the disintegration time. The upper limit is usually 50% by mass or less, preferably 19% by mass or less, and more preferably 15% by mass or less. This increases the hardness of the tablets and can improve moldability, making them less prone to cracking and chipping after molding. Therefore, the content of starch or its derivatives is usually 3 to 50% by mass, preferably 7 to 19% by mass, and more preferably 8 to 15% by mass.

[0044] Furthermore, when the food composition is in tablet form, the amount of disintegrant per tablet is usually 2 to 300 mg, preferably 2 to 200 mg, and more preferably 2 to 100 mg. The reason for setting the lower limit and upper limit is the same as for the content (mass%) mentioned above.

[0045] - Excipients - The moldability can be improved by including excipients. Examples of excipients include lactose, lactose granules, granulated sugar, mannitol, crystalline cellulose (also known as microcrystalline cellulose), magnesium carbonate, calcium carbonate, refined sucrose, glucose, hydrated glucose, low-substituted hydroxypropyl cellulose, L-cysteine, methylethyl cellulose, dextrose, sorbitol, maltodextrin, sucrose fatty acid ester, and hydroxypropyl cellulose. From the viewpoint of further improving moldability, crystalline cellulose, lactose, lactose granules, magnesium carbonate, calcium carbonate, and methylethyl cellulose are preferred.

[0046] When excipients are included, their content is usually 20% by mass or more, preferably 25% by mass or more, and more preferably 30% by mass or more, relative to the total mass of the food composition. This can further improve moldability. The upper limit is usually 80% by mass or less, preferably 70% by mass or less, and more preferably 60% by mass or less. This can prevent the tablet size from becoming too large and improve ease of administration. Therefore, the content of excipients is usually 20-80% by mass, preferably 25-70% by mass, and more preferably 30-60% by mass.

[0047] Furthermore, when the food composition is in tablet form, the amount of excipient per tablet is usually 20 to 800 mg, preferably 50 to 400 mg, and more preferably 80 to 200 mg. The reason for setting the lower limit and upper limit is the same as for the content (mass%) mentioned above.

[0048] - Fluidizing agent - By including a fluidizing agent, adhesion between powder particles can be suppressed and fluidity can be promoted. Examples of fluidizing agents include silicon dioxide (e.g., fine-grained silicon dioxide (fine-grained silicon dioxide, silica), light anhydrous silicic acid, hydrated fine-grained silicic acid, hydrated silicon dioxide), magnesium aluminometasilicate, calcium silicate, and talc. From the viewpoint of further improving powder fluidity, fine-grained silicon dioxide is preferred, and porous fine-grained silicon dioxide is more preferred.

[0049] When a fluidizing agent is included, its content is usually 1% by mass or more relative to the total mass of the food composition. This can improve manufacturability, such as the supply of powder during tableting. The upper limit is usually 3% by mass or less, preferably 2.5% by mass or less, and more preferably 2% by mass or less. This can improve moldability. Therefore, the content of the fluidizing agent is usually 1 to 3% by mass, preferably 1 to 2.5% by mass, and more preferably 1 to 2% by mass.

[0050] -lubricant- The inclusion of a lubricant reduces friction, thereby suppressing the adhesion of powder particles to the tablet press. Examples of lubricants include metal stearate salts (e.g., calcium stearate, magnesium stearate), magnesium oxide, sodium stearyl fumarate, sucrose fatty acid esters, hydrated magnesium silicate (talc), and finely powdered anhydrous silicic acid. From the viewpoint of preventing adhesion to the tablet press and suppressing tableting problems (e.g., binding, sticking, picking), calcium stearate and magnesium stearate are preferred.

[0051] When a lubricant is included, its content is usually 1% by mass or more relative to the total mass of the food composition. This helps to suppress tableting problems. The upper limit is usually 3% by mass or less, preferably 2.5% by mass or less, and more preferably 2% by mass or less. This can improve tablet hardness (making them less brittle) and reduce the likelihood of the coating layer peeling off. Therefore, the lubricant content is usually 1 to 3% by mass, preferably 1 to 2.5% by mass, and more preferably 1 to 2% by mass.

[0052] -Binder- By including a binder, the cracking and chipping of tablets can be suppressed. Examples of binders include gum arabic, gelatin, corn starch, sodium alginate, pullulan, dextrin, cyclodextrin, methylcellulose, ethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxymethylcellulose, polyethylene glycol, and casein. From the viewpoint of further suppressing the cracking and chipping of tablets, methylcellulose, ethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropylcellulose, and hydroxypropylmethylcellulose are preferred.

[0053] When a binder is included, its content is usually 0.01% by mass or more relative to the total mass of the food composition. This helps to suppress tablet breakage and chipping. The upper limit is usually 50% by mass or less. This can make delayed disintegration less likely. Therefore, the binder content is usually between 0.01% and 50% by mass.

[0054] - Surfactants - Examples of surfactants include anionic surfactants such as sodium alkyl sulfate, nonionic surfactants such as polyoxyethylene sorbitan fatty acid esters, polyoxyethylene fatty acid esters, and polyoxyethylene castor oil derivatives, and fatty acid monoglycerides.

[0055] -Colorants- Examples of colorants include tar dyes, caramel, red iron oxide, titanium dioxide, riboflavins, green tea extract, anthocyanins, chlorophyll, saffron, gardenia pigment, copper chlorophyllin sodium, food colorants such as Yellow No. 5, Yellow No. 4, Red No. 2, Red No. 3, Red No. 102, Blue No. 2, Blue No. 1, Yellow No. 4, orange essence, caramel, carmine, β-carotene, edible lake pigments such as aluminum lake, organic pigments such as copper chlorophyllin sodium, copper chlorophyll, and riboflavin, inorganic pigments such as ferric oxide, yellow ferric oxide, black iron oxide, zinc oxide, titanium dioxide, black iron oxide, brown iron oxide, zinc oxide, gold leaf, and medicinal charcoal, and plant and animal extracts such as licorice extract, catechu tannin powder, turmeric extract, and green tea powder.

[0056] - Flavoring agent - Examples of flavoring agents include sweeteners (e.g., saccharin sodium, dipotassium glycyrrhizinate, aspartame, stevia, thaumatin, acesulfame potassium, sucralose, sorbitol, maltitol, mannitol, reduced starch syrup, reduced palatinose, xylitol, erythritol, lactitol and other artificial sweeteners, trehalose, maple syrup), flavorings (e.g., lemon, lemon-lime, orange, vanillin, l-menthol, peppermint oil, peppermint micron X-8277-T, dry-coated matcha #421, limonene, peppermint oil, mint oil, lychee oil, orange oil, lemon oil and other plant essential oils), acidulants (e.g., citric acid, tartaric acid, malic acid, lactic acid), and green tea powder.

[0057] -Adsorbent- Examples of adsorbents include special calcium silicate (fluorite®) and cellulose derivatives.

[0058] -Antistatic agent and decay extender- Examples of antistatic agents and disintegration extenders include light anhydrous silicic acid, calcium stearate, magnesium stearate, and glyceryl monostearate.

[0059] -Film-forming components (coating agents)- The film-forming component is the component that forms the coating film that constitutes the coated tablet. Examples of film-forming components include celluloses such as hydroxypropyl methylcellulose, carmellose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxymethylcellulose, methylcellulose, ethylcellulose, hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate, and carboxymethyl ethylcellulose; natural substances such as gelatin, casein, lecithin, chitosan, shellac, pullulan, and gum arabic; carboxyvinyl polymer, povidone, crospovidone, polyvinyl alcohol, polyacrylic acid, aminoalkyl methacrylate copolymer (e.g., Eudragid® E, Eudragid® RS), methacrylic acid copolymer (e.g., Eudragid® L30-55), sucrose, polyethylene glycol, amylopectin, or titanium dioxide. In particular, celluloses and natural substances are preferred from the viewpoint of efficiently suppressing the bitterness, astringency, and odor mainly caused by hydroxytyrosol in olive extracts, improving palatability, and exhibiting the long-term stability of hydroxytyrosol, with hydroxypropyl methylcellulose, hydroxypropylcellulose, and shellac being more preferred.

[0060] -Plasticizer- Plasticizers are components that form the coating film that makes up coated tablets. Examples of plasticizers include polyhydric alcohols such as polyethylene glycol, propylene glycol, and glycerin; synthetic esters such as triacetin, triethyl citrate, and glycerin fatty acid esters; carnauba wax; sorbitol; maltitol; erythritol; and dextrin. Among these, polyhydric alcohols are preferred, and glycerin is more preferred, from the viewpoint of efficiently suppressing the bitterness, astringency, and odor mainly caused by hydroxytyrosol in olive extract, improving palatability, and exhibiting the long-term stability of hydroxytyrosol.

[0061] -Other ingredients- Other components include, for example, monosaccharides such as glucose, fructose, and lactose; polysaccharides of disaccharides or more (e.g., sucrose, maltose, xylose, isomerized lactose, oligosaccharides, sucrose, trehalose, sorbitol, mannitol, etc.); sugar alcohols (e.g., palatinite®, reduced starch hydrolysates, etc.); starch syrup; isomerized sugars; pigments such as titanium dioxide and iron oxide; colorants; flavorings; cetanol; sodium lauryl sulfate, etc. Although other examples besides physiologically active components have been explained using tablets as an example, the dosage form of the food composition of the present invention is not limited to tablets, as will be described later. For example, when the food composition is in liquid form, examples include, but are not limited to, a base (water), sugar and / or sugar alcohol (fructose-glucose liquid sugar, erythritol, etc.), acidulant (same as above), sweetener (same as above), fruit juice (apple juice, orange juice, tomato juice, etc.), antioxidant (vitamin C, etc.), coloring agent (same as above), flavoring agent (same as above), petrolatum, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water, or hard fat. Furthermore, when the food composition is in powder form to be dissolved in water when consumed, examples include, for example, an acidulant (same as above), a sweetener (same as above), an excipient (calcium phosphate, the excipient mentioned above), a coloring agent (same as above), and a flavoring agent (same as above).

[0062] Furthermore, the heat stress relief agent of the present invention can also be appropriately incorporated into other functional foods, health foods, beauty foods, nutritional functional foods, health functional foods, foods with functional claims, foods for specified health uses, quasi-drugs, pharmaceuticals, and hot food and beverages.

[0063] Furthermore, the present invention can be used in combination with other plant or animal-derived substances, extracts, or compounds that enhance the heat stress mitigation effect. [Examples]

[0064] (Example 1) "Method for preparing olive extract" Dried olive leaves were mixed with distilled water to make a 5% (w / w) solution. After 24 hours at room temperature, the mixture was roughly filtered through a cloth, then filtered through a 2.7 μm filter, and finally filtered through a 0.45 μm filter. The solid content was measured using a halogen moisture meter (Ohaus Halogen Moisture Meter MB45), and distilled water was added to obtain an olive extract with a solid content of 1% (w / w).

[0065] "Olive extract suppresses the reduction in cell count caused by heat stress." Mouse dermal fibroblasts 8x10 5 Cells were seeded at a density of cells / 6 cm plate. The following day, the culture medium was replaced with 3 mL of DMEM 5% FBS (bovine serum) or DMEM 5% FBS containing 0.00001% olive extract (solids concentration), and the cells were cultured at 37°C and 5% CO2 (normal conditions). The following day, the cells were subjected to heat treatment by standing in a 42°C air incubator for 2 or 3 hours, and then cultured back to normal conditions until the next day. The culture medium was replaced with DMEM 5% FBS containing 0.005% Neutral Red (w / v), and the cells were cultured for 2 hours. The cells were washed twice with 1 mL of PBS(-), and lysed with 500 μL of 1% acetic acid and 50% EtOH. 100 μL was added to a 96-well plate, and the amount of Neutral Red taken up by the viable cells was measured using a plate reader (molecular device SpectraMax ABS Plus, absorbance 540 nm) to determine the cell count.

[0066] (result) The relative cell count is shown with 2 hours of heat stress set as 1 (Figure 1). In the culture medium alone, the cell count decreased to approximately 90% when heat stress was applied from 2 hours to 3 hours (left half of Figure 1). On the other hand, in the culture medium containing olive extract, the cell count was similar at 42°C for 2 hours and 3 hours (right half of Figure 1), indicating that olive extract can suppress the decrease in cell count due to heat stress.

[0067] (Example 2) "Olive extract suppresses the decrease in cell count and increase in melanin production caused by heat stress." Nakazawa et al. reported that heat stress suppresses the proliferation of melanocytes (melanin-forming cells) and promotes melanin production (Journal of Investigative Dermatology, Volume 110, Issue 6, June 1998, Pages 972-977). Olive extract suppressed the decrease in cell number induced by heat stress in mouse fibroblasts (Figure 1). Therefore, we analyzed whether olive extract suppresses melanin production during heat stress using melanin-forming cells.

[0068] Mouse B16 melanoma cells 2 x 10 4 Cells were seeded at a density of cells / 6 cm plate. The following day, the culture medium was replaced with 3 mL of DMEM 5% FBS or DMEM 5% FBS containing 0.00001% olive extract (solids), and cultured under normal conditions. From the next day, heat treatment was performed by placing the cells in a 40°C air incubator for 1 hour daily. The culture medium was changed every two days, and the cell count and melanin content were measured after 6 days. Melanin content was measured according to previously reported methods (evaluation method of melanin production inhibitors using B16 melanoma cells, Moisturizing, whitening, anti-wrinkle, antioxidant evaluation and experimental method manual, pp. 112-114). Cell count was measured by 0.25% (w / v) trypsin treatment and trypan blue staining, and the number of viable cells was measured using a hemocytometer. The remaining cells were centrifuged (800 rpm, 4°C, 5 min), and the cells were washed with 500 μL of PBS(-). The cells were centrifuged again, collected, and lysed with 150 μL of 1N NaOH. 100 μL was added to a 96-well plate, and the absorbance at 405 nm was measured using a plate reader.

[0069] (result) Compared to the control group (37°C), heat stress (40°C, 1 hour, 6 days) reduced the cell count by approximately 70% (Figure 2). At this time, the amount of melanin per cell increased 1.4 times, indicating that heat stress caused an increase in melanin (Figure 3). Next, heat stress was performed with and without olive extract, and the cell count and melanin amount were compared. In the olive extract-containing medium, the cell count increased by approximately 1.3 times (Figure 4), and the amount of melanin per cell decreased (Figure 5). Therefore, it was shown that olive extract can suppress the decrease in cell count and increase in melanin amount caused by heat stress.

[0070] (Example 3) The following is an example of the composition when preparing tablets.

[0071] <Tablet manufacturing> Ingredient Content (mg / grain) Olive extract of the present invention 31.0 Powdered reduced maltose syrup 60.0 Partially pregelatinized starch 30.0 Crystalline cellulose 95.6 Fine-grained silicon dioxide 5.4 Carboxymethylcellulose 4.5 Calcium stearate 4.5 Total 281 Coating agent Hydroxypropyl methylcellulose 2.7 Glycerin 0.3 Total number of coated tablets: 284

[0072] (Example 4) <Manufacturing of vanishing cream> Olive extract of the present invention 0.2 Stearic acid 5.0 Stearyl alcohol 4.0 Butyl alcohol stearate 8.0 Glycerin monostearate 2.0 Propylene glycol 10.0 Glycerin 4.0 Potassium hydroxide 0.2 Preservatives and antioxidants: appropriate amount Fragrance (appropriate amount) Purified water remainder Note that the above composition is expressed in parts by weight.

[0073] Propylene glycol, potassium hydroxide, and the olive extract of the present invention were added to purified water and dissolved, then heated and maintained at 70°C (aqueous phase). Other components were mixed and heated until melted, then maintained at 70°C (oil phase). The oil phase was gradually added to the aqueous phase, and after all of it had been added, the temperature was maintained for a while to allow the reaction to occur. After that, the mixture was uniformly emulsified with a homomixer and cooled to 30°C while stirring well to produce a whitening vanishing cream.

[0074] (Example 5) <Cold cream manufacturing> Olive extract of the present invention 0.2 Solid paraffin 5.0 Beeswax 10.0 Vaseline 15.0 Liquid paraffin 41.0 Glycerin monostearate Polyoxyethylene 2.0 Sorbitan monolaurate 2.0 Butylhydroxytoluene 0.4 Soap powder 0.1 Purified water remainder Fragrance (appropriate amount) Preservative (appropriate amount)

[0075] Soap powder, seaweed extract, and the olive extract of the present invention are added to purified water, heated and dissolved, and maintained at 70°C (aqueous phase). The remaining ingredients are mixed, heated and melted, and maintained at 70°C (oil phase). The oil phase is gradually added to the aqueous phase while stirring, and after all of it has been added, the temperature is maintained for a while to allow the reaction to occur. Then, it is uniformly emulsified with a homomixer and cooled to 30°C while stirring well to produce a whitening cold cream.

[0076] (Example 6) <Manufacturing of emulsion> Olive extract of the present invention 0.2 Microcrystalline wax 1.0 Beeswax 2.0 Lanolin 20.0 Liquid paraffin 10.0 Squalane 5.0 Sorbitan sesquioleate 4.0 Polyoxyethylene (20 moles) Sorbitan monooleate 1.0 Propylene glycol 7.0 Purified water remainder Fragrance (appropriate amount) Preservatives and antioxidants: appropriate amount

[0077] The following steps were performed: 1. Add polyethylene glycol, triethanolamine, and the olive extract of the present invention to purified water, heat and dissolve, and maintain the temperature at 70°C (aqueous phase). 2. Mix in the other components, heat and melt, and maintain the temperature at 70°C (oil phase). 3. Add the oil phase to the aqueous phase for preliminary emulsification, then uniformly emulsify using a homomixer. After emulsification, the mixture is cooled to 30°C while stirring well to produce a whitening emulsion.

[0078] (Example 7) <Manufacturing of lotions> (Alcohol phase) 95% ethyl alcohol 10.0 Polyoxyethylene hydrogenated castor oil 2.0 Propylene glycol 0.1 Oleyl alcohol 0.1 Lecithin 2.5

[0079] (aqueous phase) Olive extract of the present invention 0.2 Glycerin 5.0 Purified water remainder UV absorber (appropriate amount)

[0080] After mixing and dissolving the components of the alcohol phase and the aqueous phase, a whitening lotion was produced by solubilizing the aqueous phase and the alcohol phase separately.

[0081] (Example 8) <Manufacturing of peel-off type packaging> (Alcohol phase) 95% ethanol 10.0 Polyoxyethylene (15 moles) Oleyl alcohol ether 2.0 Preservative (appropriate amount) Fragrance (appropriate amount)

[0082] (aqueous phase) Olive extract of the present invention 0.2 Polyvinyl alcohol 12.0 Glycerin 3.0 Polyethylene glycol 1500 1.0 Purified water remainder

[0083] Prepare the aqueous phase by mixing and dissolving the components of the aqueous phase in purified water at 80°C, and then cool it to 50°C. Prepare the aqueous phase by mixing and dissolving each component of the alcohol phase in polyvinyl alcohol at room temperature. Add the alcohol phase to the aqueous phase, mix until uniform, and then allow to cool to produce the whitening powder. We manufactured a Ruoff-type pack.

[0084] (Example 9) <Manufacturing of whitening skin lotion> (A) component Ethanol 10.0 Monolauric acid Polyoxyethylene (20 mol) sorbitan 5.0 Dibutylhydroxytoluene 0.01 Fragrance 0.05

[0085] (B) Component Olive extract of the present invention 0.2 Glycerin 5.0 Xanthan gum 0.1 Hydroxyethylcellulose 0.1 Purified water remainder

[0086] The above-mentioned component (A) was mixed and uniformly dissolved, then component (B) was added and stirred to disperse it, and then the mixture was filled into a container to produce a whitening skin lotion.

[0087] (Example 10) <Manufacturing of sunscreen cream> (A) component Lipophilic glyceryl monostearate 0.1 Polyoxyethylene cetyl ether 0.1 Polyoxyethylene tetraoleate Sorbitol 0.05 Behenyl alcohol 5.0 Triethylhexanoin 7.0 Ethylhexyl methoxycinnamate 6.0 Oxybenzone 2.0

[0088] (B) Component Xanthan gum 0.3 Dilauroyl glutamate lysine Sodium solution 0.1 1,3-Butylene glycol 0.1 Olive extract of the present invention 0.2 Purified water remainder

[0089] After stirring components (A) and (B) separately to 80°C, component (A) was added to component (B) while stirring with a homomixer (5000 rpm) at 80°C. Stirring continued at 80°C for 5 minutes. After that, the mixture was cooled to room temperature while stirring to produce sunscreen cream.

[0090] (Example 11) <Manufacturing of cleansing gel> (A) component Olive extract of the present invention 0.2 Sodium dilauroyl glutamate solution 0.3 Glycerin 20.0 Sorbitol 3.5 Purified water remainder

[0091] (B) Component Liquid paraffin 47.0 Isononyl isononanoate 15.0 Methylpolysiloxane 5.0 Decamethylcyclopentasiloxane 5.0 Polyoxyethylene triisostearate Lenglyceryl 2.0

[0092] (A) Stir the ingredients at 40°C for 5 minutes (800 rpm), then stir at room temperature for 15 minutes (800 rpm). (B) Add the ingredients at a rate of approximately 1 g / min while stirring (800 rpm) for the first 10 minutes, and then add at a rate of approximately 3 g / min while stirring (800 rpm). Defoaming treatment was performed three times to produce the cleansing gel.

[0093] (Example 12) <Manufacturing of cleansing milk> (A) component Olive extract of the present invention 0.2 Glycerin 7.5 Glycerin fatty acid ester 0.5 Purified water remainder

[0094] (B) Component Polyglyceryl diisostearate 7.0 Polyglyceryl distearate 3.0 Caprylic / Capric Triglyceride 55.5

[0095] (C) Component Xanthan gum 0.05 Purified water 4.95

[0096] (A) is heated and dissolved at 80°C, and then (B) is heated and dissolved at 80°C and mixed and stirred using a homomixer (6000 rpm). After that, it is cooled (to about 35°C), and then (C) is added while stirring with a paddle to produce the cleansing milk.

[0097] (Example 13) <Manufacturing of beauty serums> (A) component 1,3-Butylene Glycol 5.0 Glycerin 5.0 Xanthan gum 0.5 Hydroxyethylcellulose 0.5 Sodium hyaluronate 0.2

[0098] (B) Component Preservative (appropriate amount) Chelating agent 0.1 Purified water remainder

[0099] (C) Component Olive extract of the present invention 0.2 Sodium citrate (appropriate amount) Citric acid (appropriate amount)

[0100] (A) The other ingredients are dispersed by stirring in 1,3-butylene glycol and glycerin. (B) Purified water is heated to 70°C, and the chelating agent and preservative are stirred and dissolved, then cooled to about 40°C. After that, ingredients (A) are mixed into ingredients (B) and stirred. After confirming that ingredients (A) and (B) have been stirred and dissolved, the pH is adjusted with ingredient (C) to produce the beauty serum.

[0101] (Example 14) <Manufacturing of beauty supplements> (product raw material) Olive extract of the present invention 0.2 Beeswax 5.0 Glycerin fatty acid ester 2.0 γ-tocopherol 3.0 Olive oil remaining

[0102] The raw materials were mixed and encapsulated in a gelatin film to produce soft capsules for beauty supplements.

[0103] (Example 15) (Manufacturing of capsules) The following mixture was enclosed in soft capsules to produce capsules. Olive extract of the present invention 30 mg Hyaluronic acid 150mg Maltose 50mg Reduced maltose 50mg Pine bark extract 20mg Silicon dioxide 10mg Vitamin E 0.1mg Niacin 0.5mg Asparagine 0.25mg Aspartic acid 0.25 mg Magnesium 1 mg [Industrial applicability]

[0104] This invention can be used in the cosmetics industry and the medical industry.

Claims

1. A heat stress reliever containing olive extract as its active ingredient.

2. A heat stress mitigator according to claim 1, wherein heat stress is characterized by a decrease in cell count.

3. A heat stress reliever according to claim 1, wherein heat stress is caused by an increase in melanin production.

4. A heat stress mitigation composition comprising the agent described in any one of claims 1 to 3.

5. A heat stress-relieving composition, wherein the composition is a cosmetic, quasi-drug, pharmaceutical, or food / beverage.

6. A method for alleviating heat stress, comprising the step of administering olive extract.

7. The method for mitigating heat stress according to claim 6, wherein heat stress is characterized by a decrease in the number of cells.

8. The method for mitigating heat stress according to claim 6, wherein heat stress is characterized by an increase in melanin production.

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