JP3553477B2 - Active oxygen scavenging enzyme activity promoter - Google Patents

Description

【００３６】
製造例６〜１５についても同様の試験を行い、同様のＳＯＤおよびカタラーゼ活性の促進作用を示した。以上の実験例１の結果において、特にメハジキ及びジオウの抽出物が活性酸素消去酵素の活性促進作用が強かった。また、上記のいずれの抽出物も安定性に優れていた。
【００３７】
【発明の効果】
以上のことから、本発明のメハジキを含有する活性酸素消去酵素の活性促進剤、またはメハジキ、カンゾウ、ジオウ、ロクジョウの抽出物を一種又は二種以上含有するカタラーゼ活性促進剤は安定性に優れ、優れた活性酸素消去酵素の活性促進作用を示した。これらは、皮膚のみならず、活性酸素消去酵素が絡むあらゆる臓器についても同様である。[0001]
BACKGROUND OF THE INVENTION
TECHNICAL FIELD The present invention relates to an activity promoter for active oxygen scavenging enzyme, and in particular, active oxygen scavenging enzyme effective for preventing oxidation of living bodies including skin by promoting the activity of active oxygen scavenging enzyme which has been reduced by ultraviolet rays or aging. It relates to an activity promoter.
[0002]
[Prior art]
The skin is located in the outermost layer of the living body, is an organ in which active oxygen is easily generated due to the influence of ultraviolet rays and the like, and is constantly exposed to the oxygen stress. On the other hand, active oxygen scavenging enzymes exist in the skin cells, and protect the skin cells from injury of active oxygen unless active oxygen exceeding that capacity is generated. However, it is known that the activity of scavenging reactive oxygen scavenging enzymes in skin cells decreases with aging, and when the injury due to active oxygen surpasses its protective response, the skin is oxidized and the cellular function deteriorates and ages. It is thought to go. Accordingly, active oxygen scavengers and antioxidants have been studied as disclosed in JP-A-9-118630 and JP-A-9-208484 for the purpose of protecting against active oxygen injury, and active oxygen scavenging enzymes such as SOD and catalase, SOD Foods, cosmetics, quasi-drugs or pharmaceuticals containing active oxygen scavengers and antioxidants such as active substances have been developed.
[0003]
[Problems to be solved by the invention]
However, active oxygen scavenging enzymes such as SOD and catalase have problems in stability, and many active oxygen scavengers and antioxidants are not effective enough, so they are excellent in stability and highly effective in scavenging active oxygen. There is a need for enzyme activity promoters.
[0004]
[Means for Solving the Problems]
Under such circumstances, as a result of intensive investigations to obtain an active oxygen scavenging enzyme activity promoter excellent in protective action against active oxygen injury, the present inventors have found that extracts of Japanese pheasant, licorice, elephant and rouge are The inventors have found that the activity of the active oxygen scavenging enzyme inherent in the skin cells is stable and the present invention has been completed.
[0005]
That is, the present invention is an active oxygen scavenging enzyme activity promoter containing an extract of Japanese pheasant, licorice, dianthus, and rhododendron .
[0006]
The pheasant used in the present invention (scientific name: Leonurus sibiricus L.) is a herb belonging to the genus Lamiaceae, and grows naturally in sunny places in Yamano from Honshu, Shikoku, Kyushu, Korea and China to Southeast Asia. The whole plant of the territory of Mehajiki is the name of Yakumosou, and is used as a herbal medicine for postpartum hemostasis, menstrual irregularities, dizziness and abdominal pain.
[0007]
The licorice used in the present invention (scientific name: Glycyrrhiza uralensis) is a herb belonging to the genus Leguminosae and is distributed from Africa, Europe to Asia, Australia and western North America. Roots and running stems (strons) are used in Chinese medicine as licorice. You may use closely related licorice (scientific name: Glycyrrhiza glabra).
[0009]
Giant (scientific name: Rehmannia glutinosa Liboschitz) used in the present invention is a herb belonging to the genus Giantiaceae, and is distributed in North China and Korea. What dried the roots of Jiou is called ground yellow and is used as a herbal medicine.
[0010]
Locust used in the present invention is a deer family of Mandeka deer (scientific name: Cervus (Cervus) elephus L. var. Xanthhopygus Milne-Edwards) and Manshu deer (scientific name: Cervus (Sika) nippin t. It is a young horn (bag angle) that is not cornified yet or slightly keratinized.
[0011]
The extract of Japanese pheasant used in the present invention is extracted from a part or whole plant of a plant such as a leaf, stem, bark, flower, fruit, root, etc. Preferably, the one obtained by extracting from the whole plant of the above-ground part of a pheasant used as a crude drug is good. The extract of licorice is extracted from a part of the plant body such as leaves, stems, bark, flowers, fruits, roots or whole plants of licorice. Preferably, licorice roots and strons used as herbal medicines are extracted from dried ones. Diou extract is extracted from a part of the plant body such as leaves, stems, bark, flowers, fruits, roots, or the whole grass. Preferably, the dried roots of sage used as a crude drug are good. The extract of Rokujou is extracted from the horn of Manshu deer and Manchu deer. The preparation method is not particularly limited, and for example, it may be extracted by heating or may be extracted at room temperature. In addition, some of the above extracts are commercially available, and these can be used.
[0012]
Examples of the solvent to be extracted include water, lower monohydric alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, Propylene glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, Propyl ether, etc.). Water, lower monohydric alcohol and liquid polyhydric alcohol are preferable, and water, ethanol, 1,3-butylene glycol and propylene glycol are particularly preferable. These solvents may be used alone or in combination of two or more.
[0013]
In addition, the extract of Japanese pheasant, licorice, elephant, and rhododendron may be used as it is, or may be used after being subjected to treatments such as concentration, dilution, and filtration as necessary. Further, the extracted solution may be subjected to a treatment such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.
[0014]
The active oxygen scavenging enzyme activity promoter of the present invention may use the above extract as it is, and within the range that does not impair the effects of the extract of pheasant, licorice, territory, and rhododendron , ordinary foods, cosmetics, pharmaceuticals Fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, antiseptics, perfumes, moisturizers, powders Components such as a body, an ultraviolet absorber, a thickener, a dye, an antioxidant, a whitening agent, a chelating agent, an excipient, a stabilizer, a preservative, a binder, and a disintegrant can also be blended.
[0015]
The active oxygen scavenging enzyme activity promoter of the present invention can be used in any of foods, cosmetics, quasi-drugs, and pharmaceuticals. Examples of the dosage form include tablets, beverages, lotions, creams, and emulsions. , Gel, aerosol, ointment, poultice, paste, plaster, essence, pack, detergent, bath preparation, foundation, dusting, lipstick, powder, pill, tablet, injection, suppository, emulsion, capsule Agents, granules, liquids (including tinctures, fluid extracts, spirits, suspensions, limonades, etc.).
[0016]
【Example】
Next, in order to describe the present invention in detail, examples of production and experimental examples of the extract used in the present invention are given, but the present invention is not limited thereto.
[0017]
Production Example 1 Hot water extract of Japanese pheasant 400 mL of purified water was added to 20 g of the whole plant of the terrestrial pheasant, extracted at 95-100 ° C. for 2 hours, filtered, the filtrate was concentrated, freeze-dried, Of hot water extract was obtained.
[0018]
Production Example 2 Hot water extract of licorice 400 mL of purified water was added to 20 g of dried licorice root, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated, freeze-dried and dried with licorice. 1.0 g of hot water extract was obtained.
[0020]
Production Example 4 Geothermal hot water extract 400 g of purified water was added to 20 g of dried geous root, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated, lyophilized and dried. 1.2 g of hot water extract was obtained.
[0021]
Production Example 5 Extract from hot water of Antarcticus 400 mL of purified water is added to 20 g of dried sachet of Manchu deer, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate is concentrated and freeze-dried. 0.3 g of hot water extract of Rokujou was obtained.
[0022]
Manufacture example 6: Extract of a Japanese reptile, added to 900 g of dried dry vegetative herb, and extracted at room temperature for 7 days, filtered, and concentrated to dryness. .5 g was obtained.
[0023]
Production Example 7 Licorice ethanol extract To 100 g of dried licorice whole plant, 900 mL of ethanol was added, extracted at room temperature for 7 days, filtered, the filtrate was concentrated to dryness, and licorice ethanol extract 6 0.0 g was obtained.
[0025]
Production Example 9 Diou ethanol extract 900 g of ethanol was added to 100 g of a dried product of Diou whole plant, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness to obtain an ethanol extract of Diou. 0.2 g was obtained.
[0026]
Production Example 10 Ethanol extract of Rokujou Add 900 mL of ethanol to 100 g of dried sardine of Manchua deer, extract for 7 days at room temperature, filter, concentrate the filtrate to dryness, 0.8g was obtained.
[0027]
Production Example 11 Extract from 1,3-butylene glycol aqueous solution of pheasant fish 1 kg of purified water and 1 kg of 1,3-butylene glycol were added to 100 g of dried terrestrial herb whole grass, extracted for 7 days at room temperature, and then filtered. As a result, 1.9 kg of a 1,3-butylene glycol aqueous solution extract of swordfish was obtained.
[0028]
Production Example 12 Extract of 1,3-butylene glycol aqueous solution of licorice 1 kg of purified water and 1 kg of 1,3-butylene glycol were added to 100 g of dried licorice root, extracted for 7 days at room temperature, filtered, and licorice 1.9 kg of 1,3-butylene glycol aqueous solution extract was obtained.
[0030]
Production Example 14 Diou 1,3-butylene glycol aqueous solution extract 1 kg of purified water and 1 kg of 1,3-butylene glycol were added to 100 g of dried diou root, and extracted at room temperature for 7 days. 1.9 kg of a 1,3-butylene glycol aqueous solution extract was obtained.
[0031]
Production Example 15 1,3-Butyleneglycol aqueous solution extract of Aspergillus added 1 kg of purified water and 1 kg of 1,3-butyleneglycol to 100 g of dried sachet of Manju deer, extracted for 7 days at room temperature, and then filtered. 1.9 kg of a 1,3-butylene glycol aqueous solution extract of Rokujou was obtained.
[0032]
Next, experimental examples will be given to explain the effects of the present invention in detail.
[0033]
Experimental Example 1 Promoting activity of reactive oxygen scavenging enzyme in epidermal cells 1 × 10 ⁵ mouse keratinocyte-derived Pam212 cells were seeded in a 6cm dish, and hot water extracts of Japanese pheasant , licorice, elephant and rouge were added to a concentration of 1μg / ml 10%
The cells were cultured in Eagle's MEM containing FCS for 4 days under conditions of 5% CO ₂ and 37 ° C. Next, the cells were washed twice with PBS (−), detached with a rubber policeman, and ultrasonically disrupted with 500 μl of PBS (−) as an enzyme sample solution. The SOD activity in the enzyme sample solution was measured using SOD Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). That is, 100 μl of enzyme sample solution, 450 μl of color reagent and 450 μl of enzyme solution (xanthine oxidase) or blank solution (phosphate buffer used to dissolve xanthine oxidase) were mixed and reacted at 37 ° C. for 20 minutes, and then the reaction was stopped. 1 ml of the solution was added, and the absorbance at 560 nm was measured. Catalase activity was measured using the amount of hydrogen peroxide decomposed as an index. That is, 50 μl of enzyme sample solution was added to 1 ml of 50 mM phosphate buffer containing 10 mM hydrogen peroxide, and the change in absorbance at 240 nm for 1 minute was measured. As a control, cells obtained by sonicating cells cultured without addition of a sample were used. In this experiment, it has been confirmed that the hot water extract of Japanese pheasant, licorice, terrestrial and cypress added to the medium itself does not affect the measurement results of SOD and catalase activity of the enzyme sample solution.
[0034]
These test results are shown in Table 1 with the SOD and catalase activity of the cells cultured with the addition of each sample as 100. The hot-water extracts of Japanese pheasant , licorice, Giant and Locust showed SOD and catalase activity in mouse keratinocyte-derived Pam212 cells. In particular, the hot-water extract of pheasant has a high SOD activity promoting effect, and the hot water extract of licorice, dianthus, and rhododendron has a high promoting activity of catalase activity.
[0035]
[Table 1]

[0036]
The same tests were conducted on Production Examples 6 to 15 and showed similar SOD and catalase activity promoting effects. In the results of Experimental Example 1 described above, particularly, the extract of Japanese pheasant and dianthus had a strong activity promoting effect on the active oxygen scavenging enzyme. In addition, any of the above extracts was excellent in stability.
[0037]
【The invention's effect】
From the above, the active oxygen scavenging enzyme activity promoter containing the oysterfish of the present invention , or the catalase activity promoter containing one or more extracts of the oysterfish, licorice, dianthus, and rhododendron are excellent in stability, Excellent activity of scavenging enzyme for active oxygen scavenging enzyme was shown. The same applies not only to the skin but also to all organs that involve the active oxygen scavenging enzyme.

Claims (5)

An active oxygen-scavenging enzyme activity promoter containing an extract of Japanese pheasant. The enzyme activity promoter according to claim 1, wherein the active oxygen scavenging enzyme is superoxide dismutase (SOD). The enzyme activity promoter according to claim 1, wherein the active oxygen scavenging enzyme is catalase. The enzyme activity promoter according to any one of claims 1 to 3 , wherein the extraction solvent is water, a lower monohydric alcohol and / or a liquid polyhydric alcohol. A catalase activity promoter containing one or more extracts of Japanese pheasant, licorice, dianthus, and rhododendron.