JP5085198B2 - Antioxidant - Google Patents

Description

  The present invention relates to an antioxidant, an anti-aging agent, and an anti-inflammatory agent containing an extract of Asteraceae white eyebrows, and skin cosmetics and cosmetic foods and drinks containing them.

In recent years, active oxygen has attracted attention as a factor that oxidizes biological components, and its adverse effect on living organisms has become a problem. Reactive oxygen is generated in the process of energy metabolism in living cells, and superoxide [that is, superoxide anion (· O ₂ ⁻ ) generated by one-electron reduction of oxygen molecules, hydrogen peroxide (H ₂ O ₂ ), Singlet oxygen ( ¹ O ₂ ), hydroxy radical (.OH)], and the like. Such active oxygen is essential for the phagocytic sterilization mechanism and plays an important role in removing viruses and cancer cells.
However, excessive generation of the active oxygen attacks in vivo molecules constituting membranes and tissues in the living body and induces various diseases. Superoxide, which is produced in vivo and is a starting material for other active oxygen, is usually eliminated sequentially by the catalytic action of superoxide dismutase (SOD) contained in cells. However, when the production of superoxide is excessive or when the action of SOD is reduced, the superoxide concentration becomes high due to insufficient superoxide elimination. This is one of the causes of tissue damage such as rheumatoid arthritis, Behcet's disease, myocardial infarction, stroke, cataract, spots, freckles, wrinkles, diabetes, arteriosclerosis, shoulder stiffness, coldness, skin aging, etc. It is considered. Among these, since the skin is directly affected by environmental factors such as ultraviolet rays, superoxide is likely to be generated. Therefore, when the superoxide concentration is increased, biological tissues such as collagen are decomposed, denatured, or crosslinked. Or oxidize fats and oils to produce lipid peroxides that damage cells, forming skin wrinkles, causing skin aging, inflammation, and skin pigmentation (See Non-Patent Document 1).
Therefore, attempts have been made to obtain active oxygen scavenging substances, radical scavenging substances, hydrogen peroxide scavenging substances, etc. from natural products advantageous in terms of safety, and extracts of Brassica Brassica plants (see Patent Document 1), Efficacy has been confirmed for extracts of plants belonging to the genus Ryukyubekei (see Patent Document 2), extracts of scallops (see Patent Document 3), extracts of sulfur (see Patent Document 4), and the like.

Glutathione is a tripeptide composed of three amino acids, glutamic acid, cysteine and glycine, and is a compound having a major intracellular cysteine residue. The role of glutathione in cells is radical scavenging, regulation of cell functions by redox, SH donors of various enzymes, and is also known as an antioxidant component. It is thought that the action expression of glutathione is derived from the cysteine residue. However, it has been reported that the amount of glutathione in the skin decreases with aging, which reduces the ability of the skin to oxidize and damages cellular components such as DNA and proteins. It is considered to be.
In this way, promoting the production of glutathione in the skin increases the defense against oxidative stress that declines with aging, and also suppresses damage against oxidative stress caused by ultraviolet rays, preventing skin aging, treating it, or causing stains, etc. It is considered that an improvement in pigmentation can be expected. Then, as a thing which has glutathione production promotion effect | action, the bilberry extract or walnut extract (refer patent document 5), the extract of gardenia plant (refer patent document 6), etc. are disclosed, for example.

The skin is composed of the epidermis, basement membrane, and dermis. The basement membrane is present at the boundary between the epidermis and the dermis, and plays an important role in maintaining the skin function as well as connecting the epidermis and the dermis (see Non-Patent Document 2). The main skeleton of the basement membrane has a network structure made of type IV collagen. In young skin, the proliferation of fibroblasts is active, and the interaction of the epidermis and dermis keeps the homeostasis by the action of the basement membrane, ensuring moisture retention, flexibility, elasticity, etc., and the skin is appearance It is also kept fresh with tension and gloss.
However, the growth of fibroblasts slows down due to the influence of certain external factors such as UV irradiation, significant drying of air, excessive skin washing, etc., and the ability to produce extracellular matrix. As the skin level decreases, the skin loses tension and elasticity, and exhibits aging symptoms such as rough skin wrinkles. In addition, type IV collagen, which is a main component of the basement membrane, is decomposed or altered, and the basement membrane structure is destroyed (see Non-Patent Document 3). As a result, the moisturizing function and elasticity of the skin are reduced, the exfoliation of the keratin begins to be abnormally peeled, the skin loses its tension and gloss, and exhibits aging symptoms such as roughness and wrinkles. Thus, it is known that changes associated with skin aging, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, etc. are associated with a decrease in basement membrane components and a change in basement membrane structure. Yes.

In recent years, the involvement of matrix proteases has been pointed out as a factor for inducing reduction or denaturation of dermal matrix components. Among these matrix proteases, MMP-1 (matrix metalloprotease-1) is known as an enzyme that degrades type I collagen, type III collagen, and the like, which are main components of the dermal matrix of the skin. The expression is greatly increased by the irradiation of ultraviolet rays, contributes to the decrease or degeneration of collagen by ultraviolet rays, and is considered to be a major factor such as the formation of wrinkles on the skin.
In addition, as a cause of skin aging associated with aging, secretion of estrogen, a type of female hormone, may decrease. Estrogen is deeply involved in maintaining the health of adult women, and its lack of secretion leads to various medical illnesses and can cause unwanted skin changes such as skin sensitivities, reduced elasticity and reduced moisture. It has been known.
Therefore, it promotes type IV collagen production, inhibition of MMP-1 activity, promotion of skin fibroblast proliferation, or reduction of estrogen secretion due to aging, thereby reducing skin aging such as formation of skin wrinkles and reduced elasticity. It can be prevented or treated.

There are various causes and onset mechanisms of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and various other skin diseases with rough skin. As its cause, histamine release inhibiting action, nitric oxide (NO) production inhibiting action, and cyclooxidase-2 (COX-2) are known.
The histamine release is a phenomenon in which histamine in mast cells is released outside the cell, and the released histamine causes an inflammatory reaction. For this reason, attempts have been made to prevent or treat allergic diseases and inflammatory diseases with substances that inhibit or suppress histamine release. As such histamine release inhibitors, for example, tranilast, sodium cromoglycate, baicalin, baicalein, promethazine hydrochloride and the like have been used. However, all of these substances have side effects and have problems in terms of safety.
Nitric oxide (NO) is a nitrogen oxide that causes air pollution, acid rain, and the like. In recent years, nitric oxide (NO) is a physiologically active substance that exhibits various functions in vivo, such as vascular endothelium-derived relaxing factor (EDRF), neurotransmitters, microorganisms in biological defense, and tumor cell injury factors. It has been reported. As a physiologically active substance, it is known that nitric oxide produced from macrophages protects against bacterial and viral infections. However, when a large amount of nitric oxide produced from the macrophages is biosynthesized, it is not non-toxic to the living body and causes destruction of the self-tissue, which may cause pathological conditions such as worsening inflammation, rheumatism and diabetes. . Also, large amounts of biosynthesized nitric oxide can cause vascular smooth muscle relaxation and excessive permeability, and can cause endotoxin shock due to a significant decrease in blood pressure.
Therefore, it is important to suppress excessive production of nitric oxide (NO) in inflammatory diseases. Examples of herbal medicines having an inhibitory effect on the production of nitric oxide include, for example, rosemary extract, carnosol, carnosic acid, coffee bean extract, primrose extract, auren extract, buckwheat extract, licorice extract, Inu rose extract, Senkyu extract, Tounin extract, Peonies extract, Yakuinin extract, Red grape extract (all refer to Patent Document 7), Tang Dynasty, Cod root bark, Japanese continuation, Car front, Far There have been reported pupae, licorice root, half-branches, spikelets, flower buds (see Non-Patent Document 4).

Inflammation is a complex reaction that exhibits symptoms such as redness, edema, fever, pain, and dysfunction. Microscopically, it consists of common reactions such as blood vessel reaction that causes plasma leakage, leukocyte infiltration, tissue destruction by inflammatory cells, and also causes systemic reactions involving the central nervous system such as fever and hyperalgesia. There is a case. Prostaglandins play an important role in such individual reactions of inflammation, and the production of prostaglandins during inflammation mainly involves the involvement of cyclooxygenase-2 (COX-2), an inducible cyclooxygenase. Are known. For this reason, many cyclooxygenase activity inhibitors represented by aspirin have been reported for the purpose of preventing or preventing inflammatory reactions (see Non-Patent Document 5). As plant-derived cyclooxygenase activity inhibitors, α-mangostin and γ-mangostin in mangosteen peel extract are disclosed (see Patent Document 8). Examples of compounds having a cyclooxygenase-2 activity inhibitory action include 2-phenyl-1,2-benzisoselenazole-3 (2H) -one, 2-phenyl-1,2-benzisoselenazole-3 (2H ) -One salts or hydrates of 2-phenyl-1,2-benzisoselenazol-3 (2H) -one are disclosed (see Patent Document 9).
Thus, inhibiting the release of histamine, inhibiting the production of excess nitric oxide (NO), and inhibiting the activity of cyclooxidase-2 (COX-2) can prevent or improve inflammatory diseases. It is extremely important.

  However, to date, it is a natural product that is easy to obtain, inexpensive, and highly safe, and does not adversely affect the quality of the additive in terms of taste, smell, feeling of use, etc. Antioxidants, anti-aging agents, and anti-inflammatory agents that can be widely used in cosmetics and beauty foods and drinks have not yet been provided, and there is a strong demand for prompt provision thereof.

JP 2003-81848 A JP 2005-29483 A JP 2006-321730 A JP 2007-8902 A JP 2006-241062 A JP 2006-347934 A JP 2002-87975 A JP 2002-47180 A JP 2000-16935 A Fragrance Journal Special Issue No. 14, p156 1995 Marinkovich MP et al. , J .; Cell. Biol. 1992 199: 695-703 Lavker et al. , J .; Invest. Dermatol. 1979 73: 59-66 “Wakan Journal of Pharmaceutical Sciences”, Vol. 15, p. 302-303, issued in 1998 Pharmacology Atlas (P184), translated by Takehiko Fukuhara, Bunkodo

An object of the present invention is to solve the conventional problems and achieve the following objects. That is, the present invention first has at least one of a superoxide scavenging action, a radical scavenging action, a hydrogen peroxide scavenging action, and a glutathione production promoting action, and is a natural product that is highly safe and easy to obtain raw materials. An object of the present invention is to provide a system antioxidant.
In addition, the present invention secondly has at least one of skin fibroblast proliferation action, type IV collagen production promotion action, matrix metalloproteinase-1 (MMP-1) activity inhibition action, and estrogen-like action, An object of the present invention is to provide a natural anti-aging agent that is highly safe and easily available.
Thirdly, the present invention has at least one of histamine release inhibitory action, nitric oxide (NO) production inhibitory action, and cyclooxidase-2 (COX-2) activity inhibitory action, and is highly safe. An object of the present invention is to provide a natural anti-inflammatory agent whose raw materials are easily available.
A fourth object of the present invention is to provide skin cosmetics and cosmetic foods and drinks containing at least one of the antioxidant, the anti-aging agent, and the anti-inflammatory agent of the present invention as active ingredients. And

  In order to solve the above-mentioned problems, it is a natural product that is easy to obtain, inexpensive, and highly safe, and does not adversely affect the quality of the added object in terms of taste, smell, feeling of use, etc. As a result of extensive studies by the present inventors on antioxidants, anti-aging agents, and anti-inflammatory agents that can be widely used in cosmetics and beauty foods and drinks, an extract of white eyebrows of Asteraceae (1) It has at least one of an excellent superoxide scavenging action, radical scavenging action, hydrogen peroxide scavenging action, and glutathione production promoting action, and is useful as an antioxidant. (2) Excellent skin fibroblast proliferation action Have a type IV collagen production promoting action, matrix metalloproteinase-1 (MMP-1) activity inhibiting action, and estrogen-like action, and are useful as an anti-aging agent, and (3 It has at least one of excellent histamine release inhibitory action, nitric oxide (NO) production inhibitory action, and cyclooxidase-2 (COX-2) activity inhibitory action, and is found to be useful as an anti-inflammatory agent. did.

The present invention is based on the above findings of the present inventors, and means for solving the above problems are as follows. That is,
<1> An antioxidant containing an extract of white eyebrows.
<2> The antioxidant according to <1>, having at least one of a superoxide scavenging action, a radical scavenging action, a hydrogen peroxide scavenging action, and a glutathione production promoting action.
<3> An anti-aging agent characterized by containing an extract of white eyebrows.
<4> Anti-aging according to the above <3>, which has at least one of dermal fibroblast proliferation action, type IV collagen production promotion action, matrix metalloproteinase-1 (MMP-1) activity inhibition action, and estrogen-like action It is an agent.
<5> An anti-inflammatory agent characterized by containing an extract of white eyebrows.
<6> The anti-inflammatory agent according to <5>, which has at least one of a histamine release inhibitory action, a nitric oxide (NO) production inhibitory action, and a cyclooxidase-2 (COX-2) activity inhibitory action.
<7> A skin cosmetic comprising an extract of white eyebrows according to any one of <1> to <6> as an active ingredient.
<8> A cosmetic food or drink comprising the white eyebrow extract according to any one of <1> to <6> as an active ingredient.

According to the antioxidant of the present invention, through at least one of excellent superoxide scavenging action, radical scavenging action, hydrogen peroxide scavenging action, and glutathione production promoting action, preventing or improving in vivo oxidation and skin aging can do.
According to the anti-aging agent of the present invention, the skin can be passed through at least one of excellent skin fibroblast proliferation action, type IV collagen production promotion action, matrix metalloproteinase-1 (MMP-1) activity inhibition action, and estrogen-like action. It is possible to prevent or improve wrinkles and skin elasticity reduction.
According to the anti-inflammatory agent of the present invention, inflammatory diseases are prevented through at least one of an excellent histamine release inhibitory action, nitric oxide (NO) production inhibitory action, and cyclooxidase-2 (COX-2) activity inhibitory action. Thorough improvement.
Further, the antioxidant, anti-aging agent, and anti-inflammatory agent of the present invention are natural extracts that are excellent in safety and do not adversely affect the quality of the addition target in terms of taste, smell, feeling of use, etc. Therefore, it is suitable for blending in skin cosmetics or adding to cosmetic foods and drinks.

(Antioxidants, anti-aging agents, and anti-inflammatory agents)
The antioxidant, anti-aging agent, and anti-inflammatory agent of the present invention contain an extract of white eyebrow grass, and further contain other components as necessary.

The antioxidant has at least one of a superoxide scavenging action, a radical scavenging action, a hydrogen peroxide scavenging action, and a glutathione production promoting action as an antioxidant action.
The anti-aging agent has at least one of dermal fibroblast proliferation action, type IV collagen production promotion action, matrix metalloproteinase-1 (MMP-1) activity inhibition action, and estrogen-like action as an anti-aging action. ing.
The anti-inflammatory agent has at least one of histamine release inhibitory action, nitric oxide (NO) production inhibitory action, and cyclooxidase-2 (COX-2) activity inhibitory action as an anti-inflammatory action.
The details of the substance having at least one of antioxidative action, anti-aging action, and anti-inflammatory action in the white brow grass extract are unknown, but the white brow grass extract has these excellent actions. So far, it has not been known at all to be useful as an antioxidant, an anti-aging agent, and an anti-inflammatory agent, and these are new findings by the inventors' diligent research.

The white eyebrow is a plant belonging to the family Asteraceae, scientific name is Gerberia piloselloides Gerbera , and it is a perennial herb that is also called Mao Dycho and Michiko. The rhizome is thick and thick. There is fluff. The leaves come out from the base and have a slight pattern, approximately circular to oval, 5 cm to 8 cm in length, and 15 to 30 cm in length and sometimes 40 cm in length.
The white eyebrows are wild around the sun, mountain slopes, roadsides, and rice fields in Jiangsu, Zhejiang, Sichuan, Guangxi, Yunnan, etc. Is readily available from
The white eyebrows are known to have such effects as coughing, sweating, diuresis, and nutritional tonic.

  The white eyebrow extract can be easily obtained by a method generally used for plant extraction. The white eyebrow extract includes a white eyebrow extract, a dried product obtained by drying a diluted solution of the extract, and a crude product or a purified product thereof.

  There is no restriction | limiting in particular as an extraction raw material of the said white eyebrow grass, According to the objective, it can select suitably, For example, the leaf part of a white eyed grass, a flower (bud) part, a branch part, a seed, a bark (these are above-ground parts) Can be used). Among these, a ground part such as a leaf part is particularly preferable.

  The white eyebrow that is the raw material for extraction can be obtained by drying or pulverizing the raw material as it is or using a crusher and subjecting it to solvent extraction. Drying may be performed in the sun or using a commonly used dryer. In addition, you may use the said white eyebrows as an extraction raw material, after giving pretreatments, such as a degreasing | defatting, by nonpolar solvents, such as hexane and benzene. In addition, by performing pretreatments such as degreasing, extraction processing of white eyebrow grass with a polar solvent can be performed efficiently.

As the solvent used for the extraction, it is preferable to use water, a hydrophilic organic solvent, or a mixed solvent thereof at a temperature from room temperature to the boiling point of the solvent.
Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, as well as water that has been subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. The water that can be used as the extraction solvent includes purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

Examples of the hydrophilic organic solvent include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene glycol, propylene glycol, Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycerin, and a mixed solvent of these hydrophilic organic solvents and water can be used.
In addition, when using the mixed solvent of the said water and a hydrophilic organic solvent, in the case of a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, and in the case of a lower aliphatic ketone, 10 masses of water. It is preferable to add 1 to 40 parts by mass with respect to parts. In the case of polyhydric alcohol, it is preferable to add 1 part by mass to 90 parts by mass with respect to 10 parts by mass of water.

  In the present invention, it is not necessary to employ a special extraction method for extracting a substance having at least one of an antioxidative action, an anti-aging action, and an anti-inflammatory action from white eyebrows which is an extraction raw material, and it is room temperature or reflux heating. Below, it can be extracted using any extraction device.

  Specifically, in the treatment tank filled with the extraction solvent, the above-ground part of white eyebrow grass as an extraction raw material is added, and further allowed to stand for 30 minutes to 2 hours with occasional stirring as necessary. After elution, the solid matter is removed by filtration, and the extract is obtained by evaporating the extraction solvent from the resulting extract and drying. The amount of the extraction solvent is usually 5 to 15 times (mass ratio) of the extraction raw material. The extraction conditions are usually about 1 to 4 hours at 50 to 95 ° C. when water is used as the extraction solvent. Moreover, when the mixed solvent of water and ethanol is used as an extraction solvent, it is normally 30 minutes-about 4 hours at 40 to 80 degreeC. In addition, the extract obtained by extracting with a solvent can be used as it is as the antioxidant, anti-aging agent, and anti-inflammatory agent of the present invention as long as the extraction solvent is highly safe.

  In order to obtain a dilute or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof, the obtained white birch grass extract is diluted, concentrated, dried, purified, etc. according to a conventional method. You may perform the process of.

  In addition, although the extract of white eyebrows obtained can be used as it is as an antioxidant, anti-aging agent, and anti-inflammatory agent, a concentrated solution or a dried product thereof is easier to use. In obtaining a dried extract, a carrier such as dextrin or cyclodextrin may be added to improve hygroscopicity. In addition, since the extract of white eyebrows has a characteristic odor, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. Since it is not used in large quantities when it is added to food and cosmetic foods and drinks, there is no practical problem even if it is not purified. The purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like.

  The white eyebrows extract obtained as described above has a superoxide scavenging action, hydrogen peroxide scavenging action, radical scavenging action, glutathione production promoting action, skin fibroblast proliferation action, type IV collagen production promoting action, matrix metallo Protease-1 (MMP-1) activity inhibitory action, estrogen-like action, histamine release inhibitory action, nitric oxide (NO) production inhibitory action, and cyclooxidase-2 (COX-2) activity inhibitory action Therefore, based on these actions, it can be used as an antioxidant, an anti-aging agent, and an anti-inflammatory agent of the present invention.

The antioxidant action of the antioxidant of the present invention is exhibited based on at least one of a superoxide scavenging action, a hydrogen peroxide scavenging action, a radical scavenging action, and a glutathione production promoting action.
The anti-aging action of the anti-aging agent of the present invention is based on at least one of a dermal fibroblast proliferation action, a type IV collagen production promoting action, a matrix metalloproteinase-1 (MMP-1) activity inhibiting action, and an estrogen-like action. It is demonstrated.
The anti-inflammatory action of the anti-inflammatory agent of the present invention is exhibited based on at least one of histamine release inhibitory action, nitric oxide (NO) production inhibitory action, and cyclooxidase-2 (COX-2) activity inhibitory action. .
Since the extract of white eyebrows of the present invention has at least one of an excellent antioxidative action, anti-aging action, and anti-inflammatory action, and is excellent in the feeling of use and safety when applied to the skin, It is suitable for blending into the skin cosmetic of the present invention described below.
In addition, it has been confirmed that the extract of white eyebrows of the present invention has at least one of an excellent antioxidant action, anti-aging action, and anti-inflammatory action and is not digested in the digestive tract. In particular, it is suitable for blending into the cosmetic food and drink of the present invention described below.

(Skin cosmetic)
The skin cosmetic of the present invention contains at least one of the antioxidant, the anti-aging agent, and the anti-inflammatory agent of the present invention as an active ingredient, and further other components appropriately selected as necessary. It contains.

  Here, the use of the skin cosmetic is not particularly limited and can be appropriately selected from various uses. For example, ointment, cream, emulsion, lotion, pack, jelly, lip balm, lipstick, bath additive, astringent , Etc.

  The blending amount of the antioxidant, the anti-aging agent, or the anti-inflammatory agent with respect to the entire skin cosmetic can be adjusted as appropriate depending on the type of skin cosmetic, physiological activity of the extract, etc. It is preferably 0.0001% by mass to 10% by mass, and more preferably 0.001% by mass to 1% by mass in terms of the extract.

The skin cosmetics can be added with various main ingredients and auxiliaries usually used in the production of the skin cosmetics and other components as long as they do not impair the objects and effects of the present invention. .
The other components are not particularly limited as long as they do not interfere with the antioxidant, anti-aging, and anti-inflammatory effects of the present invention, and may include components appropriately selected according to the purpose. , Bactericides, antibacterial agents, ultraviolet absorbers, humectants, cell activators, fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, and the like. These components may act synergistically when used in combination with the white blossom extract, resulting in superior effects than expected.

  The skin cosmetic of the present invention has high safety when used on the skin, and has an excellent superoxide scavenging action, hydrogen peroxide scavenging action, radical scavenging action, glutathione production promoting action, skin fibroblast proliferation action, Type IV collagen production promoting action, matrix metalloproteinase-1 (MMP-1) activity inhibiting action, estrogen-like action, histamine release inhibiting action, nitric oxide (NO) production inhibiting action, and cyclooxidase-2 (COX-2) It effectively exhibits at least one of its activity-inhibiting action, and is useful for in vivo oxidation prevention, aging prevention, and prevention or treatment of inflammatory diseases.

(Beauty food and drink)
The cosmetic food / beverage product of the present invention contains at least one of the antioxidant, the anti-aging agent, and the anti-inflammatory agent of the present invention as an active ingredient. Contains ingredients.
Here, the above-mentioned beauty foods and drinks are those that are less likely to harm human health and are taken orally or by digestive tract administration in normal social life. It is not limited to categories such as quasi-drugs, and includes, for example, foods that include a wide range of foods that are taken orally, such as general foods, health foods, health functional foods, quasi-drugs, and pharmaceuticals.

  The cosmetic food or drink of the present invention may be a white brow grass extract blended with any food or drink so as not to impede its activity, or a nutrient mainly composed of white brow grass extract. It may be a supplement.

  There is no restriction | limiting in particular as said beauty food-drinks, Although it can select suitably according to the objective, For example, drinks, such as a soft drink, a carbonated drink, a nutrition drink, a fruit drink, a lactic acid drink; Ice cream, ice sherbet , Shaved ice and other frozen desserts; noodles such as buckwheat, udon, harsame, gyoza skin, sushi mai, Chinese noodles, instant noodles; rice cakes, candy, gum, chocolate, tablet confectionery, snacks, biscuits, jelly, jam, cream, Sweets such as baked goods and bread; crab, salmon, clams, tuna, sardines, shrimp, skipjack, mackerel, whale, oysters, saury, squid, red sea bream, scallop, abalone, sea urchin, sea bream, tocobushi, etc .; Processed fishery and livestock products such as ham and sausage; Dairy products such as processed milk and fermented milk; salad oil, tempura oil, margarine, mayonnaise, Fats and processed foods such as yotoning, whipped cream, dressing, etc .; seasonings such as sauces, sauces; curry, stew, oyakodon, rice cake, miscellaneous cooking, Chinese rice cakes, tempura, eel rice cake, hayashi rice, oden, marbodorf, Retort pouch foods such as beef bowl, meat sauce, egg soup, omelet rice, dumplings, shumai, hamburger, meatballs; various forms of health foods and nutritional supplements; pharmaceuticals such as tablets, capsules, drinks, troches, etc. Examples include foreign products.

As said other component, the auxiliary | assistant raw material or additive normally used in manufacturing the said cosmetics food / beverage products is mentioned.
The raw material or additive is not particularly limited and may be appropriately selected depending on the intended purpose.For example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn syrup, lactose, citric acid, Tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, gum arabic , Carrageenan, casein, gelatin, pectin, agar, vitamin Bs, nicotinamide, calcium pantothenate, amino acids, calcium salts, pigments, fragrances, preservatives, and the like.

  The addition amount of the antioxidant, the anti-aging agent, or the anti-inflammatory agent of the present invention in the beauty food or drink varies depending on the type of the beauty food or drink to be used and cannot be specified unconditionally. However, what is necessary is just to add in the range which does not impair the original taste of beauty food / beverage products, 0.001 mass%-50 mass% are preferable with respect to various object cosmetic food / beverage products, 0.01 mass%-20 mass% are preferable. More preferred. Further, in the case of a cosmetic food or drink in the form of granules, tablets or capsules, 0.01% by mass to 100% by mass is preferable, and 5% by mass to 100% by mass is more preferable.

  The cosmetic food and drink according to the present invention can be taken orally on a daily basis, and has at least one of an antioxidative action, an anti-aging action, and an anti-inflammatory action depending on the action of an extract of white eyebrow which is an active ingredient. Can be exhibited extremely effectively.

  In addition, although the antioxidant, anti-aging agent, anti-inflammatory agent, skin cosmetics, and cosmetics for food and beverage of the present invention are suitably applied to humans, their respective effects are exhibited. As long as it is applicable to animals other than humans.

  Examples of the present invention will be described below, but the present invention is not limited to these examples.

(Production Example 1)
-Production of water extract of white eyebrows-
As a raw material for extraction, 100 g of ground product of white eyebrow grass was put into 1,000 mL of water, kept at 80 ° C. for 2 hours with gentle stirring, and then filtered. The filtrate was concentrated under reduced pressure at 40 ° C. and further dried with a vacuum dryer to obtain an extract (powder). The yield of the obtained extract is shown in Table 1.

(Production Example 2)
-Production of 50% by weight ethanol extract of white eyebrows-
As a raw material for extraction, 100 g of ground material of white eyebrow grass was added to 1,000 mL of 50 mass% ethanol (mass ratio of water to ethanol of 1: 1) and kept at 80 ° C. for 2 hours with gentle stirring. Then, it filtered. The filtrate was concentrated under reduced pressure at 40 ° C. and further dried with a vacuum dryer to obtain an extract (powder). The yield of the obtained extract is shown in Table 1.

(Production Example 3)
-Production of ethanol extract of white eyebrows-
As a raw material for extraction, 100 g of ground product of white eyebrow grass was put into 1,000 mL of ethanol, kept at 80 ° C. for 2 hours with gentle stirring, and then filtered. The filtrate was concentrated under reduced pressure at 40 ° C. and further dried with a vacuum dryer to obtain an extract (powder). The yield of the obtained extract is shown in Table 1.

Example 1
-Superoxide elimination test (NBT method)-
Each extract of Production Examples 1 to 3 was used as a sample, and the superoxide scavenging action was tested by the following test method.
3 mmol / L xanthine, 3 mmol / L EDTA, 1.5 mg / mL bovine serum albumin (BSA) solution, 0.75 mmol / L nitroblue tetrazolium (NBT) 0.1 mL, and 0.05 mol / L Na 2.4 mL of ₂ CO ₃ buffer (pH 10.2) was placed in a test tube, 0.1 mL of each sample solution was added thereto, and the mixture was allowed to stand at 25 ° C. for 10 minutes. Subsequently, the xanthine oxidase solution was added, it stirred rapidly, and it left still at 25 degreeC for 20 minutes. Thereafter, 0.1 mL of 6 mmol / L copper chloride was added to stop the reaction, and the absorbance at a wavelength of 560 nm was measured. The absorbance measured at this time was defined as “absorbance upon addition of sample solution and enzyme solution”.
The same operation and measurement of absorbance were performed without adding the enzyme solution. The absorbance measured at this time was defined as “absorbance when the sample solution was added and no enzyme solution was added”.
The same measurement was performed when distilled water was added without adding the sample solution. The absorbance measured at this time was defined as “absorbance when no sample solution was added and enzyme solution was added”.
Moreover, the same measurement was performed when distilled water was added without adding the enzyme solution and without adding the sample solution. The absorbance measured at this time was defined as “absorbance when no sample solution was added and no enzyme solution was added”.
And the superoxide elimination rate was calculated | required by following Numerical formula 1 from the measurement result.
<Formula 1>
Superoxide erasure rate (%) = {1- (AB) / (CD)} × 100
Where A is the absorbance when the sample solution is added and the enzyme solution is added, B is the absorbance when the sample solution is added and the enzyme solution is not added, C is the absorbance when the sample solution is not added and the enzyme solution is added, and D is The absorbance when no sample solution is added and when no enzyme solution is added is shown.

  Next, the sample concentration was decreased stepwise to measure the superoxide erasure rate, and the sample concentration (μg / mL) at which the superoxide erasure rate was 50% was determined by interpolation. The results are shown in Table 2.

表２の結果から、製造例１〜３の白眉草抽出物が、高いスーパーオキサイド消去作用を有することが確認できた。

From the results in Table 2, it was confirmed that the white eyebrow extract of Production Examples 1 to 3 had a high superoxide scavenging action.

(Example 2)
-Hydrogen peroxide elimination test-
Using each extract of Production Examples 1 to 3 as a sample, the hydrogen peroxide scavenging action was tested by the following test method.
After adding 10 μL of each sample solution to 10 μL of a standard solution of hydrogen peroxide (concentration 1.5 mmol / L) and incubating at 37 ° C. for 20 minutes, a coloring reagent [DA-64 (manufactured by Wako Pure Chemical Industries, Ltd.) was added at 10 mmol / L, 1 mL of peroxidase solution (100 units / mL, manufactured by Wako Pure Chemical Industries, Ltd.) is added to 0.1 mol / L PIPES buffer (pH 7.0) containing 0.5% by mass of Triton X-100, and the total amount is adjusted to 100 mL. Adjusted] After 2.98 mL was added and incubated at 37 ° C. for 5 minutes, the absorbance at a wavelength of 727 nm was measured.
The same operation and absorbance measurement as described above were performed without adding a standard solution of hydrogen peroxide. Further, the same measurement as described above was performed when distilled water was added without adding the sample solution.
And the elimination rate of hydrogen peroxide was calculated | required by following Numerical formula 2 from the obtained measurement result.
<Formula 2>
Hydrogen peroxide elimination rate (%) = {1− (A−B) / (C−D)} × 100
Where A is the absorbance when hydrogen peroxide standard solution is added and sample solution is added, B is the absorbance when hydrogen peroxide standard solution is not added and sample solution is added, and C is the hydrogen peroxide standard solution added and sample. Absorbance when no solution is added, and D indicates absorbance when no hydrogen peroxide standard solution is added and when no sample solution is added.

  Next, the hydrogen peroxide erasure rate was measured by gradually reducing the sample concentration, and the sample concentration (μg / mL) at which the hydrogen peroxide erasure rate was 50% was determined by interpolation. The results are shown in Table 3.

表３の結果から、製造例１〜３の白眉草の抽出物が、高い過酸化水素消去作用を有することが確認できた。

From the results in Table 3, it was confirmed that the white eyebrow extract of Production Examples 1 to 3 had a high hydrogen peroxide scavenging action.

(Example 3)
-Radical scavenging test for DPPH-
Using each extract of Production Examples 1 to 3 as a sample, radical scavenging action was tested using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), which is a very stable radical, by the following test method.
3 mL of each sample solution was added to 3 mL of a 1.5 × 10 ⁻⁴ mol / L DPPH ethanol solution, and the container was immediately sealed, shaken and allowed to stand for 30 minutes, and then the absorbance at a wavelength of 520 nm was measured.
As a control, the same operation was performed using a solvent in which the sample solution was dissolved instead of the sample solution, and the absorbance at a wavelength of 520 nm was measured. Further, as a blank, after adding 3 mL of the sample solution to ethanol, the absorbance at a wavelength of 520 nm was measured immediately.
And the radical elimination rate (%) was computed from the measurement result by following Numerical formula 3.
<Formula 3>
Radical scavenging rate (%) = {1− (BC) / A} × 100
In Formula 3, A represents the absorbance of the control, B represents the absorbance when the sample solution was added, and C represents the absorbance of the blank.

  Next, the sample concentration was decreased stepwise to measure the radical scavenging rate, and the sample concentration (μg / mL) at which the DPPH radical scavenging rate was 50% was determined by interpolation. The results are shown in Table 4.

表４の結果から、製造例１〜３の白眉草の抽出物が、高いＤＰＰＨに対するラジカル消去作用を有することが確認できた。

From the results of Table 4, it was confirmed that the white eyebrow extract of Production Examples 1 to 3 has a high radical scavenging action on DPPH.

Example 4
-Test for promoting glutathione production-
Each extract of Production Examples 1 to 3 was used as a sample, and the glutathione production promoting action was tested by the following test method.
Human normal skin fibroblasts (NB1RGB) were cultured using 10% by mass fetal bovine serum (FBS) -containing αMEM, and then cells were collected by trypsin treatment. The collected cells were diluted to 2.0 × 10 ⁵ cells / mL with α-MEM containing 10% by mass of FBS, seeded in a 48-well microplate at 200 μL per well, and cultured overnight. After culturing, 200 μL of each sample solution dissolved in D-MEM containing 1% by mass of FBS was added to each well and cultured for 24 hours. After completion of the culture, the medium was removed from each well, washed with 400 μL of PBS (−), and cells were lysed using 150 μL of M-PER (registered trademark) (manufactured by PIERCE). The total glutathione was quantified using 100 μL of this. Specifically, 100 μL of cell extract dissolved in a 96-well microplate, 50 μL of 0.1 mol / L phosphate buffer, 25 μL of 2 mmol / L NADPH, and 25 μL of glutathione reductase (final concentration 17.5 units / mL) Was added and heated at 37 ° C. for 10 minutes, 25 μL of 10 mmol / L 5,5′-dithiobis (2-nitrobenzoic acid) was added, and the absorbance at a wavelength of 412 nm until 5 minutes later was measured. Asked. The total glutathione concentration was calculated based on a calibration curve prepared using oxidized glutathione. The obtained value was corrected to the amount of glutathione per total protein.
And the glutathione production promotion rate was computed from following formula 4 from the obtained amount of glutathione. Table 5 shows the glutathione production promotion rate at a sample concentration of 200 μg / L.
<Formula 4>
Glutathione production promotion rate (%) = (B / A) × 100
In Formula 4, A represents the amount of glutathione per protein amount (control) in cells not added with the sample solution, and B represents the amount of glutathione per protein amount in cells added with the sample solution.

表５の結果から、製造例１〜３の白眉草の抽出物が、グルタチオン産生促進作用を有することが確認できた。

From the results of Table 5, it was confirmed that the white eyebrow extract of Production Examples 1 to 3 has a glutathione production promoting action.

( Reference Example 5)
-Skin fibroblast proliferation promoting effect test-
Using each extract of Production Examples 1 to 3 as a sample, the skin fibroblast proliferation promoting action was tested by the following test method.
Human normal skin fibroblasts (NB1RGB) were cultured using α-MEM containing 10% by mass of fetal bovine serum (FBS), and then collected by trypsin treatment. The collected cells were diluted with α-MEM containing 5% by mass of FBS to a concentration of 7.0 × 10 ⁴ cells / mL, then seeded at 100 μL per well in a 96-well microplate, and cultured overnight.
After completion of the culture, 100 μL of a sample solution (concentration: 400 ppm) dissolved in 5 mass% FBS-containing α-MEM was added to each well and cultured for 3 days.
The skin fibroblast proliferation effect was then measured using the MTT assay. Specifically, 100 μL of the medium was removed from each well, and MTT [3- (4,5-Dimethyl-2-thiazolyl) -2,5-diphenyl-2H dissolved in PBS (−) at a final concentration of 5 mg / mL. -Tetrazole bromide] was added to each well in an amount of 20 μL. After culturing for 4.5 hours, 100 μL of a 0.01 mol / L hydrochloric acid solution in which 10% by mass of SDS was dissolved was added to each well, and after culturing overnight, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. In addition, a blank test was performed and corrected in the same manner.
Then, from the obtained measurement results, the fibroblast proliferation promotion rate at a sample concentration of 200 μg / mL was calculated by the following formula 5. The results are shown in Table 6.
<Formula 5>
Fibroblast growth rate (%) = (St−Sb) / (Ct−Cb) × 100
In Formula 5, St is the absorbance in the cells to which the sample solution is added, Sb is the absorbance in the blank test to which the sample solution is added, Ct is the absorbance in the cells to which no sample solution is added, and Cb is not added to the sample solution. The absorbance of the blank test is shown respectively.

表６の結果から、製造例１〜３の白眉草の抽出物が、皮膚線維芽細胞増殖促進作用を有することが確認できた。

From the results shown in Table 6, it was confirmed that the extract of white eyebrows of Production Examples 1 to 3 had a skin fibroblast proliferation promoting action.

( Reference Example 6)
-Type IV collagen production promoting action test-
Using each extract of Production Examples 1 to 3 as a sample, the type IV collagen production promoting effect was tested as follows.
Human normal fibroblasts (NB1RGB) were cultured using Dulbecco's MEM medium containing 10% by mass fetal bovine serum (FBS), and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco's MEM medium to a cell density of 1.6 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well microplate, and cultured overnight. After completion of the culture, the medium was removed, 150 μL of each sample solution (sample concentration: 100 μg / mL) dissolved in 0.25% by mass of FBS-containing Dulbecco MEM medium was added to each well, and cultured for 3 days. After completion of the culture, the amount of type IV collagen in the medium in each well was measured by ELISA.
And from the obtained measurement result, the IV type collagen production promotion rate (%) was computed by following Numerical formula 6. Table 7 shows the results of the type IV collagen production promotion rate at the sample concentration of 100 μg / mL.
<Formula 6>
Type IV collagen production promotion rate (%) = (A / B) × 100
In Formula 6, A represents the amount of type IV collagen when the sample solution was added, and B represents the amount of type IV collagen when no sample solution was added.

表７の結果から、製造例１〜３の白眉草の抽出物が、ＩＶ型コラーゲン産生促進作用を有することが確認できた。

From the results in Table 7, it was confirmed that the white eyebrow extract of Production Examples 1 to 3 had a type IV collagen production promoting effect.

( Reference Example 6)
-Matrix metalloproteinase-1 (MMP-1) activity inhibition test-
Using the extracts of Production Examples 1 to 3 as samples, matrix metalloproteinase-1 (MMP-1) activity inhibitory action was tested as follows. This test method is a partial modification of the Wunsch and Heidrich method.
In a test tube with a lid, 50 μL of each sample solution dissolved in 0.1 mol / L Tris-HCl buffer (pH 7.1) containing 20 mmol / mL calcium chloride, 50 μL of MMP-1 solution, and 400 μL of the substrate solution were mixed. And incubated at 37 ° C. for 30 minutes. Next, 1 mL of a 25 mmol / L citric acid solution was added to stop the reaction, 5 mL of ethyl acetate was added, and the mixture was shaken vigorously. This was centrifuged (1600 × g, 10 minutes), and the absorbance of the ethyl acetate layer at a wavelength of 320 nm was measured. The same enzyme reaction and absorbance measurement as described above were performed by adding an equal amount of buffer solution to the sample solution instead of the sample solution. Furthermore, in each case, Tris-HCl buffer was added instead of the MMP-1 solution, and the same operation and measurement were performed.
In addition, as the MMP-1 solution, collagenase Type IV (manufactured by Sigma) was dissolved in a buffer and diluted to 0.1 mg / mL.
As a substrate solution, Pz-peptide (manufactured by BACHEM Fenchemikaline AG) was dissolved in 0.1 mol / L Tris-HCl buffer containing 20 nmol / L calcium chloride so as to have a concentration of 0.5 mol / L. used.
And from the obtained measurement result, MMP-1 activity inhibition rate was computed by following formula 7.
<Formula 7>
MMP-1 activity inhibition rate (%) = {1− (C−D) / (A−B)} × 100
In Formula 7, A is the absorbance at a wavelength of 320 nm when the sample solution is not added and the enzyme is added, B is the absorbance at a wavelength of 320 nm when the sample solution is not added and the enzyme is not added, and C is the sample solution added and the enzyme Absorbance at a wavelength of 320 nm with addition, D represents the absorbance at a wavelength of 320 nm with addition of a sample solution and without addition of an enzyme, respectively.

  Next, the MMP-1 activity inhibition rate was measured by gradually reducing the sample concentration, and the sample concentration (μg / mL) that inhibits MMP-1 activity by 50% was determined by interpolation. The results are shown in Table 8.

表８の結果から、製造例１〜３の白眉草の抽出物が、マトリックスメタロプロテアーゼ−１（ＭＭＰ−１）活性阻害作用を有することが確認できた。

From the results shown in Table 8, it was confirmed that the extracts of white eyebrows of Production Examples 1 to 3 had a matrix metalloproteinase-1 (MMP-1) activity inhibitory action.

( Reference Example 8)
-Estrogen-like action test-
Using the extracts of Production Examples 1 to 3 as samples, estrogen-like action was tested by the following test method.
Human breast cancer-derived cells (MCF-7) were cultured using MEM containing 10% by mass fetal bovine serum (FBS), 1% by mass NEAA, and 1 mmol / L sodium pyruvate, and then treated with trypsin. Was recovered. 3.0 × 10 ⁴ cells using phenol red-free MEM (T-MEM) containing 10% by mass of FBS, 1% by mass of NEAA, and 1 mmol / L sodium pyruvate treated with activated carbon. After dilution to a concentration of / mL, the cells were seeded at 450 μL per well in a 48-well microplate and cultured to establish the cells. Six hours later (day 0), 50 μL of each sample solution prepared to 10 times the final concentration with T-MEM was added to each well, and the culture was continued. On the third day, the medium was removed, 500 μL of the sample solution prepared to the final concentration with T-MEM was added to each well, and the culture was further continued.
Estrogen-like effects were measured using the MTT assay. After completion of the culture, the medium was removed, and MTT [3- (4,5-Dimethyl-2-thiazolyl) dissolved in MEM containing 1% by mass of NEAA and 1 mmol / L sodium pyruvate at a final concentration of 0.4 mg / mL. ) -2,5-diphenyl-2H-tetrazolium Bromide] was added to each well in an amount of 200 μL. After culturing for 2 hours, blue formazan produced in the cells was extracted with 200 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. As a positive control, 10 ⁻⁹ mol / L estradiol was used.
Then, from the obtained measurement results, an estrogen-like action (estrogen-dependent proliferation action) was calculated from Equation 8 below. Table 9 shows the results of estrogen-like effects at a sample concentration of 50 μg / mL.

<Formula 8>
Estrogen-like action rate (%) = (A / B) × 100
In Equation 8, A represents the absorbance when the sample solution is added. B represents the absorbance when no sample solution is added.

表９の結果から、製造例１〜３の白眉草の抽出物が、エストロゲン様作用を有することが確認できた。

From the results of Table 9, it was confirmed that the white eyebrow extract of Production Examples 1 to 3 had an estrogenic action.

( Reference Example 9)
-Histamine release inhibition (hexosaminidase release inhibition) action test-
Each extract of Production Examples 1 to 3 was used as a sample, and the histamine release inhibitory action was tested by the following test method. Since intracellular histamine is released and hexosaminidase is released at the same time, the histamine release inhibitory action can be evaluated using hexosaminidase release as an index.
1.0 × 10 ⁶ rat basophil leukemia cells (RBL-2H3 cells) were added to a medium (15% by mass fetal bovine serum (FBS) -added S-MEM medium; the same applies hereinafter) placed in a 25 mL culture flask. Seeded and cultured for 4 days at 37 ° C., 5% CO ₂ -95% air. Cells were then collected by trypsinization and centrifugation (800 rpm, 4 minutes). The obtained cells were suspended in the medium at 4.0 × 10 ⁵ cells / mL, and mouse monoclonal anti-dinitrophenyl group IgE (DNP-Specific IgE) was added thereto at a concentration of 0.5 μg / mL. This cell suspension was seeded in 100 μL per well of a 96-well microplate and cultured at 37 ° C. under 5% CO ₂ -95% air for 24 hours. After completion of the culture, the medium in each well was removed and washed twice with Silaganian buffer.
Next, 30 μL of the buffer solution and 10 μL of each sample solution were added and incubated at 37 ° C. for 10 minutes. Subsequently, 10 μL of dinitrophenylated bovine serum albumin (DNP-BSA) was added and incubated at 37 ° C. for 15 minutes. Thereafter, 10 μL of the supernatant was transferred to a new 96-well microplate under ice cooling, and 10 μL of 1 mmol / L p-nitrophenyl-N-acetyl-β-D-glucosamide solution was added thereto, and the mixture was incubated at 37 ° C. for 1 hour. Incubated. After completion of the reaction, 250 μL of a 0.1 mol / L Na ₂ CO ₃ —NaHCO ₃ solution was added, and the absorbance A at 415 nm was measured with a microplate reader at 650 nm as a control. The same treatment and absorbance measurement were performed for the cell supernatant to which the Siraganian buffer was added instead of the sample solution (the absorbance measured at this time was designated as B). Absorbance measurement was also performed on the cell supernatant and 0.1 mol / L Na ₂ CO ₃ —NaHCO ₃ solution reacted in the same manner (the absorbance measured at this time was C). The same operation was performed for the sample in which a Siraganian buffer was added instead of DNP-BSA (the absorbance measured at this time was designated as D).
And the hexosaminidase release suppression (histamine release suppression) rate was computed from the obtained measurement result by the following numerical formula 9.

<Formula 9>
Inhibition rate of hexosaminidase release (%) =
[1-{(ACD) / (BD)}] × 100
However, in the said Numerical formula 9, AD represents the same meaning as the description in the said test method.

  Next, the concentration of each hexosaminidase release is measured by decreasing each sample concentration step by step, and the sample concentration (μg / mL) that inhibits hexosaminidase release by 50% is determined by interpolation. It was. The results are shown in Table 10.

表１０の結果から、製造例１〜３の白眉草の抽出物が、ヒスタミン遊離抑制（ヘキソサミニダーゼ遊離抑制）作用を有することが確認できた。

From the results of Table 10, it was confirmed that the extracts of white eyebrows of Production Examples 1 to 3 have histamine release inhibition (hexosaminidase release inhibition) action.

( Reference Example 10)
-Nitric oxide (NO) production inhibitory action test-
Each extract of Production Examples 1 to 3 was used as a sample, and the nitric oxide (NO) production inhibitory action was tested by the following test method.
Mouse macrophage cells (RAW264.7) were cultured using Dulbecco's MEM containing 10% by mass fetal bovine serum (FBS), and then the cells were collected with a cell scraper. The collected cells were diluted with 10% by mass of FBS-containing phenol red-free Dulbecco MEM to a concentration of 3.0 × 10 ⁶ cells / mL, and then seeded at 100 μL per well in a 96-well microplate. Incubate for hours. After completion of the culture, the medium was removed, 100 μL of each sample solution dissolved in 10% by mass of FBS-containing phenol red-free Dulbecco MEM containing DMSO having a final concentration of 2% by mass was added to each well, and the final concentration was 1 μg / mL. 50 μL of lipopolysaccharide (LPS, E. coli 0111; B4, manufactured by DIFCO) dissolved in mass% FBS-containing phenol red-free Dulbecco MEM was added and cultured for 48 hours. The amount of NO production was measured using the amount of nitrite ion (NO ²⁻ ) as an index. After completion of the culture, the same amount of grease reagent (1% by mass of sulfanilamide, 0.1% by mass of N-1-naphthyl ethylenedihydride in 5% by mass of phosphoric acid solution) was added to the culture solution in each well, Reacted for 10 minutes at room temperature. After the reaction, absorbance at a wavelength of 540 nm was measured. Based on the amount of nitric oxide (NO) produced by the control, the NO production inhibition rate was calculated from the following formula 10.

<Formula 10>
NO production inhibition rate (%) = {1− (A−B) / (C−D)} × 100
In Formula 10, A is the absorbance at a wavelength of 540 nm when a sample is added and stimulated with LPS, B is the absorbance at a wavelength of 540 nm when the sample is added and LPS is not stimulated, C is the absorbance at a wavelength of 540 nm when LPS is stimulated for control, and D Represents the absorbance at a wavelength of 540 nm when LPS was not stimulated as a control.

Next, the concentration of each sample solution was decreased stepwise to measure the NO production inhibition rate, and the concentration IC _{50 at} which the NO production inhibition rate was 50% was determined by interpolation (this IC ₅₀ value is small). NO production suppression effect is so strong). The results are shown in Table 11.

表１１の結果から、製造例１〜３の白眉草の抽出物が、高い一酸化窒素（ＮＯ）産生抑制作用を有することが認められた。

From the result of Table 11, it was recognized that the extract of the white eyebrows of manufacture examples 1-3 has a high nitric oxide (NO) production inhibitory effect.

( Reference Example 10)
-PGE ₂ production inhibitory action test via cyclooxygenase (COX-2)-
Using each extract of Production Examples 1 to 3 as a sample, cyclooxygenase (COX-2) activity inhibitory action was tested by the following test method. This test is described in Hwang, B .; -Y. The method (Planta Medica 67 (2001), 406-410) was modified with some modifications.
First, after pre-culturing mouse-derived macrophage-like cells (RAW264.7), the cells were collected, prepared to 2 × 10 ⁵ cells / mL, and seeded at 100 μL each in a 96-well microplate, at 37 ° C., 5% CO _2. For 18 hours. After culturing, in order to acetylate and inactivate COX-1 already present and COX-2 expressed in a small amount, the culture medium was replaced with 500 μmol / L aspirin-containing medium and cultured for 4 hours. The cells were washed three times with PBS (−), 200 μL of each sample solution dissolved in a medium containing 1 μg / mL lipopolysaccharide (LPS) was added, and cultured for 18 hours. After completion of the culture, PGE ₂ in the supernatant was quantified using PGE ₂ EIA Kit (Cayman, Chemical, Ann Arbor, MI, USA), and each sample solution was taken as 100% to give PGE ₂ production. The inhibition rate was calculated.

Next, the concentration _{IC 50} for each sample the concentration of the solution stepwise reduced measuring PGE ₂ production inhibition rate through the COX-2, PGE ₂ production inhibition rate through the COX-2 is 50% Was determined by interpolation (the smaller the IC ₅₀ value, the stronger the inhibition of PGE ₂ production via COX-2). The results are shown in Table 12.

表１２の結果から、製造例１〜３の白眉草の抽出物が、ＣＯＸ−２を介したＰＧＥ_２産生抑制作用を有することが認められた。

From the results of Table 12, it was confirmed that the white eyebrow extract of Production Examples 1 to 3 has a PGE ₂ production inhibitory action via COX-2.

(Formulation Example 1)
-Emulsion-
An emulsion was prepared from the following composition by a conventional method.
-Water extract of white eyebrow grass of production example 1 ... 0.10 g
・ Jojoba oil: 4.00 g
・ 1,3-Butylene glycol ・ ・ ・ 3.00g
・ Polyoxyethylene cetyl ether (20E.O.) ... 2.50 g
・ Olive oil ... 2.00g
・ Squalane ... 2.00g
・ Cetanol ... 2.00g
・ Glyceryl monostearate ... 2.00g
・ Oleic acid polyoxyethylene sorbitan (20E.O.) ... 2.00g
・ Methyl paraoxybenzoate 0.15 g
・ Twilight extract ... 0.10g
・ Dipotassium glycyrrhizinate ... 0.10g
・ Ginkgo biloba extract ... 0.10g
・ Conchiolin ... 0.10g
・ Oat extract ... 0.10g
・ Chicken extract ... 0.10g
・ Fragrance ... 0.05g
・ Purified water: remainder (total 100.00 g)

(Formulation Example 2)
-Lotion-
A lotion was produced from the following composition by a conventional method.
-50% by weight ethanol extract of white eyebrow grass of Production Example 2 ... 0.10 g
・ Glycerin ... 3.00g
・ 1,3-Butylene glycol ・ ・ ・ 3.00g
・ Oleic acid polyoxyethylene sorbitan (20E.O.) ... 2.00g
・ Methyl paraoxybenzoate 0.15 g
・ Citric acid ... 0.10g
・ Sodium citrate: 0.10 g
・ Oil-soluble licorice extract ... 0.10g
・ Seaweed extract ... 0.10g
・ Cudin extract ... 0.10g
・ Xylobiose Mixture ... 0.05g
・ Fragrance ... 0.05g
・ Purified water: remainder (total: 100.00 g)

(Formulation Example 3)
-Cream-
A cream was produced from the following composition by a conventional method.
-Ethanol extract of white eyebrows in Production Example 3 ... 0.10 g
・ Squalane ... 10.00g
・ 1,3-butylene glycol ... 6.00g
・ Liquid paraffin ・ ・ ・ 5.00g
・ Salah beeswax 4.00 g
・ Cetanol ... 3.00g
・ Glyceryl monostearate ... 3.00 g
・ Lanoline ... 2.00g
・ Oleic acid polyoxyethylene sorbitan (20E.O.) ... 1.50 g
・ Methyl paraoxybenzoate ... 1.50g
・ Stearic acid: 1.00 g
・ Yeast extract ... 0.10g
・ Perilla extract ... 0.10g
-Linden extract ... 0.10g
・ Jiuyu extract ... 0.10g
・ Fragrance ... 0.10g
・ Purified water: remainder (total: 100.00 g)

(Formulation Example 4)
−Pack−
A pack was produced from the following composition by a conventional method.
-Water extract of white eyebrow grass of Production Example 1 0.20 g
・ Polyvinyl alcohol: 15.00g
・ Ethanol ... 10.00g
・ Propylene glycol: 7.00 g
・ Polyethylene glycol ... 3.00g
・ Sage extract ... 0.10g
・ Toki extract ... 0.10g
・ Carrot extract ... 0.10g
・ Ethyl paraoxybenzoate 0.05g
・ Fragrance ... 0.05g
・ Purified water: remainder (total: 100.00 g)

(Formulation Reference Example 5)
-Tablet dietary supplements-
The following mixture was tableted to produce a tablet-shaped dietary supplement.
・ Water extract of white eyebrow grass of Production Example 1 30g
・ Powdered sugar (sucrose) 178g
・ Sorbit ・ ・ ・ 10g
・ Glycerin fatty acid ester: 12g

(Formulation Reference Example 6)
-Granular dietary supplement-
The following mixture was formed into granules to produce a dietary supplement.
-50% by weight ethanol extract of white eyebrow grass of Production Example 2 20 g
・ Beat oligosaccharide ... 1000g
・ Vitamin C ... 167g
・ Stevia extract ... 10g

(Formulation Reference Example 7)
-Granular dietary supplement-
The following mixture was formed into granules to produce a dietary supplement.
-Ethanol extract of white eyebrow grass of production example 3 ... 20 g
・ Beat oligosaccharide ... 1000g
・ Vitamin C ... 167g
・ Stevia extract ... 10g

A skin cosmetic containing at least one of the antioxidant, anti-aging agent, and anti-inflammatory agent of the present invention has at least one of an excellent antioxidant effect, anti-aging effect, and anti-inflammatory effect, Effective in preventing oxidation in the body, preventing and improving skin wrinkles and skin elasticity, preventing rough skin, and preventing rough skin, such as ointments, creams, emulsions, lotions, packs, jellies, lip balms, lipsticks, bathing Widely used in agents, astringents, etc.
In addition, the cosmetic food and drink to which at least one of the antioxidant, the anti-aging agent, and the anti-inflammatory agent of the present invention is added has at least one of an anti-oxidation action, an anti-aging action, and an anti-inflammatory action that are excellent even when taken orally. Since it has any of them and is excellent in safety, it is widely used, for example, in health foods and nutritional supplements.

Claims (2)

  An antioxidant comprising an extract of white eyebrows.   The antioxidant according to claim 1, which has at least one of a superoxide scavenging action, a radical scavenging action, a hydrogen peroxide scavenging action, and a glutathione production promoting action.