JP2016204272A - Active oxygen species remover, as well as skin external preparation for removing active oxygen species, health foods, and goods for daily life - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide active oxygen species removers, skin external preparations for removing active oxygen species, health foods for removing active oxygen species, and goods for daily life for removing active oxygen species, which are used for antiaging and the like and have excellent active oxygen species-removal action with less side effect and high safety for a human body.SOLUTION: The present invention provides an active oxygen species remover containing Sapindaceae Dodonaea viscosa plant extract as an active ingredient. Preferably, the active oxygen species is at least one selected from the group consisting of super oxide, singlet oxygen, hydroxy radical, hydroperoxy radical, and nitric oxide. The invention also provides a skin external preparation for removing active oxygen species, a health food for removing active oxygen species, goods for daily life for removing active oxygen species which contain the active oxygen species remover.SELECTED DRAWING: None

Description

  The present invention relates to a reactive oxygen species removing agent, a skin external preparation for removing reactive oxygen species, a health food for removing reactive oxygen species, and a household sundries for removing reactive oxygen species. Specifically, the present invention comprises an active oxygen species remover, an active oxygen species removing skin external agent, an active oxygen species removing health food, an active oxygen species, and an excellent active ingredient comprising an extract of Sindaceae dodonea viscosa. It relates to household goods for seed removal.

  The active oxygen species are superoxide, hydroxy radical, hydrogen peroxide and singlet oxygen in a narrow sense, but also include nitric oxide and lipid peroxide in a broad sense. Aerobic organisms produce energy necessary for life support by consuming oxygen in the electron transport system in mitochondria, but reactive oxygen species are always generated during the metabolic process. It is well known that living organisms use reactive oxygen species when leukocytes take in foreign substances such as bacteria. However, when reactive oxygen species are generated excessively by external factors such as ultraviolet rays and chemical substances, reactive oxygen species cause non-specific reactions such as DNA damage and lipid peroxidation, and circulation such as arteriosclerosis. It causes many diseases such as organ diseases, dermatological diseases such as atopic dermatitis, diabetes and cancer. Furthermore, from a cosmetic point of view, reactive oxygen species are thought to be involved in skin aging such as spots, buckwheat and wrinkles. The living body has superoxide dismutase, an enzyme that removes superoxide, and catalase, an enzyme that removes hydrogen peroxide, against reactive oxygen species that have both merits and demerits. However, it cannot be said that it has sufficient activity to eliminate the excessively produced reactive oxygen species. Therefore, a search for a substance having an antioxidant ability has been actively conducted. Among the active oxygen species in the narrow sense described above, superoxide is an active oxygen species that is first generated when an oxygen molecule captures an unpaired electron, and then is sequentially converted into a hydroxy radical and hydrogen peroxide by reduction. That is, superoxide is a precursor of other reactive oxygen species, and its removal is very important.

  However, other than superoxide dismutase which is an enzyme specific to superoxide, only substances such as vitamin C and bilirubin (Non-patent Document 1) are known to remove superoxide. Therefore, it can be said that the development of a superoxide remover that can be taken orally or by application is very important.

Debit E. Barenano (David E. Baranano), 3 other authors, “Bilirubin reductase: A major physicothetology of the world. (10) Biological reductase: A major physicothetology. Sun, Vol. 99, No. 25, pp. 16093-16098

  An object of the present invention is to provide a reactive oxygen species removing agent, an active oxygen species removing external preparation for active oxygen species removal, an active oxygen species, which are excellent in reactive oxygen species removing action, have few side effects, and have high safety to the human body. It is to provide health food for removing seeds and household goods for removing active oxygen species.

  The inventors of the present invention are excellent in the action of removing specific oxygen species such as superoxide, singlet oxygen, hydroxy radical, hydroperoxy radical, and nitric oxide, particularly a reactive oxygen species removing enzyme. It has been found that it has superoxide dismutase (SOD) activity and that anti-aging effects can be expected by removing reactive oxygen species, and at the same time, it is safe for living bodies, and the present invention has been completed.

  The present invention includes the following aspects (1) to (5).

(1) A reactive oxygen species remover comprising, as an active ingredient, a plant extract of Sapindaceae Dodonaea viscosa.

(2) The reactive oxygen species removal according to (1), wherein the reactive oxygen species is at least one selected from the group consisting of superoxide, singlet oxygen, hydroxy radical, hydroperoxy radical, and nitric oxide. Agent.

(3) A skin external preparation for removing reactive oxygen species, comprising the reactive oxygen species removing agent according to (1) or (2).

(4) A health food for removing reactive oxygen species, comprising the reactive oxygen species removing agent according to (1) or (2).

(5) A household item for removing active oxygen species, comprising the active oxygen species removing agent according to (1) or (2).

  ADVANTAGE OF THE INVENTION According to this invention, it is excellent in a reactive oxygen species removal effect | action, there are few side effects, and the safety to a human body is high, the reactive oxygen species removal agent for the purpose of anti-aging, the skin external preparation for active oxygen species removal, active It is possible to provide health foods for removing oxygen species and household goods for removing active oxygen species.

(1) Plant extract The active ingredient of the reactive oxygen species removing agent of the present invention is an extract of Sindaceaeae Dodonaea viscosa (hereinafter sometimes simply referred to as “plant extract”).

  The site | part used in the said plant is not specifically limited, It can extract from a flower, a root, a leaf, a branch, a stem, etc. The plant extract is usually obtained by subdividing a dried plant as necessary, immersing it in a solvent, and filtering it. The degree of fragmentation and the fragmentation method are not particularly limited, and examples thereof include a method of pulverizing a dried plant body by a conventionally known method to form a powder.

  Examples of the solvent used for the immersion treatment include water, a hydrophilic organic solvent such as ethanol, propylene glycol, and 1,3-butylene glycol, an organic solvent such as dichloromethane, or a mixture thereof. Specific examples of the mixed solvent include a water / ethanol mixed solution (50% ethanol, 70% ethanol, etc.), and a water / butylene glycol mixed solution (50% butylene glycol, etc.). The temperature of the solvent in the immersion treatment is not particularly limited and is usually treated at room temperature (22 to 25 ° C.), but the solvent can be boiled and extracted by refluxing. The immersion time is not particularly limited, but is usually about 1 day to 1 week. The form of the plant extract thus obtained may be any of powder, gel and paste.

(2) Reactive oxygen species remover The plant extract can be used as an active oxygen species remover as it is. In particular, it has an excellent removal action on at least one selected from the group consisting of superoxide, singlet oxygen, hydroxy radical, hydroperoxy radical, and nitric oxide as the active oxygen species.

(3) Reactive oxygen species removing skin external preparation The active oxygen species removing agent of the present invention can be used as it is or blended into a skin external preparation and used as a reactive oxygen species removing skin external preparation.
The blending amount of the plant extract in the active oxygen species removing skin external preparation is not particularly limited, but is 0.0001 based on the dry solid weight and the total amount of the active oxygen species removing skin external preparation. -10.0% by weight is preferable, more preferably 0.0005-5.0% by weight, and particularly preferably 0.01-5% by weight. When the blending amount is not less than the lower limit, the effect of the present invention is sufficiently obtained. On the other hand, when the blending amount is less than the upper limit, the effect corresponding to the increase tends to be recognized.

  Although the dosage form of the skin external preparation for removing reactive oxygen species of the present invention is not particularly limited, for example, it can be a dosage form such as lotions, emulsions, creams, ointments, packs and the like.

  In addition, the skin external preparation for removing reactive oxygen species of the present invention can be used in preparations for pharmaceuticals, quasi-drugs, cosmetics and the like as needed in addition to the plant extract as long as the effects of the present invention are not impaired. Components and additives used, for example, coloring matters (coloring agents), preservatives, surfactants, fragrances, pigments and the like can be appropriately blended as optional components.

  Such optional ingredients include, for example, macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, liquid lanolin, hydrogenated coconut oil Oils such as hardened oil, mole, castor oil, beeswax, candelilla wax, carnauba wax, ibotarou, lanolin, reduced lanolin, hard lanolin, jojoba wax, mineral oil (mineral oil), waxes, liquid paraffin, squalane, pristane, Hydrocarbons such as ozokerite, paraffin, ceresin, petrolatum, microcrystalline wax;

  Cetyl isooctanoate, isopropyl myristate, hexyl decyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, Di-2-heptylundecanoic acid glycerin, tri-2-ethylhexanoic acid glycerin, tri-2-ethylhexanoic acid trimethylolpropane, triisostearic acid trimethylolpropane, tetra-2-ethylhexanoic acid pentane erythritol, hydrogenated coco Esters such as glyceryl, polyglyceryl stearate, methyl paraoxybenzoate (methylparaben);

  Linear polysiloxanes such as dimethylpolysiloxane, methylphenylpolysiloxane, diphenylpolysiloxane, cyclic polysiloxanes such as octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, dodecamethylcyclohexanesiloxane, amino-modified polysiloxane, polyether-modified poly Oil agents such as silicone oil such as siloxane, alkyl-modified polysiloxane, and modified polysiloxane such as fluorine-modified polysiloxane;

  Fatty acid soap (sodium laurate, sodium palmitate, etc.), anionic surfactants such as potassium lauryl sulfate, alkylsulfuric triethanolamine ether, cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, laurylamine oxide , Imidazoline-based amphoteric surfactants (such as 2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt), betaine surfactants (such as alkylbetaines, amidebetaines, sulfobetaines), acyls Amphoteric surfactants such as methyl taurine, sorbitan fatty acid esters (such as sorbitan monostearate, sorbitan sesquioleate), glycerin fatty acids (such as glyceryl monostearate), propylene glycol Fatty acid esters (propylene glycol monostearate, etc.), hardened castor oil derivatives, glycerin alkyl ethers, sucrose fatty acid esters, surfactants such as nonionic surfactants such as alkyl glucoside;

  Polyhydric alcohols such as polyethylene glycol, 1,3-butylene glycol and glycerine; higher fatty acids such as stearic acid; moisturizing ingredients such as sodium pyrrolidonecarboxylate, lactic acid and sodium lactate; guar gum, quince seed, carrageenan, galactan, Gum arabic, pectin, mannan, starch, curdlan, methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, methylhydroxypropylcellulose, chondroitin sulfate, dermatan sulfate, glycogen, heparan sulfate, sodium hyaluronate, tragacanth gum, keratan sulfate, chondroitin, mucoitin sulfate, Hydroxyethyl guar gum, carboxymethyl guar gum, dextran, keratosulfate, locust bean gum, succinoglucan Caronic acid, chitin, chitosan, carboxymethylchitin, agar, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer (carbomer), polyacrylic acid such as sodium polyacrylate and / or its salt, polyethylene glycol, bentonite, fine particle cellulose gel, etc. A thickener;

  Powders such as mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, anhydrous silicic acid (silica), aluminum oxide, barium sulfate whose surface may be treated;

  Bengara, yellow iron oxide, black iron oxide, cobalt oxide, ultramarine, bitumen, titanium oxide, zinc oxide inorganic pigments that may be treated on the surface; titanium mica, fish phosphorus that may be treated on the surface Pearls such as foil and bismuth oxychloride; red 202, red 228, red 226, yellow 4, blue 404, yellow 5, red 505, red 230, which may be raked, Organic pigments such as Red No. 223, Orange No. 201, Red No. 213, Yellow No. 204, Yellow No. 203, Blue No. 1, Green No. 201, Purple No. 201, Red No. 204;

  Organic powders such as polyethylene powder, polymethyl methacrylate, nylon powder, organopolysiloxane elastomer;

  Paraaminobenzoic acid UV absorbers, anthranilic acid UV absorbers, salicylic acid UV absorbers, cinnamic acid UV absorbers, benzophenone UV absorbers, sugar UV absorbers, 2- (2'-hydroxy-5 ' UV absorbers such as -t-octylphenyl) benzotriazole, 4-methoxy-4'-t-butyldibenzoylmethane;

  Lower alcohols such as ethanol and isopropanol; vitamin B such as vitamin A or a derivative thereof, vitamin B6 hydrochloride, vitamin B6 tripalmitate, vitamin B6 dioctanoate, vitamin B2 or a derivative thereof, vitamin B12, vitamin B15 or a derivative thereof; Preferred examples include vitamins such as α-tocopherol, β-tocopherol, γ-tocopherol, and vitamin E acetate, vitamins D, vitamin H, pantothenic acid, panthetin, pyrroloquinoline quinone, and the like.

  The reactive oxygen species removing agent of the present invention can be used by blending in oral preparations such as supplements and nutrients in addition to the above-mentioned skin external preparation for removing reactive oxygen species.

  The external preparation for removing active oxygen species of the present invention can be applied to the skin to remove active oxygen species from the skin and prevent inflammation, cancer, aging, lifestyle-related diseases and the like caused by the active oxygen species. .

(4) Health food The active oxygen species removing agent of the present invention can be used as a health food for removing active oxygen species by containing it in food. The food containing the active oxygen species removing agent of the present invention is not particularly limited, and for example, meat, fish, vegetables, eggs, confectionery, noodles, soybeans, breads, rices, soups, sauces, sauces, and gyoza , Wonton, sweet potato, spring roll, Chinese bun, fried food, croquette, hamburger, tempura, pizza, takoyaki, okonomiyaki, gratin, sprinkle, honey.

  The method for containing the active oxygen species remover of the present invention in food is not particularly limited. For example, a method of mixing with a food material, a method of immersing food in a solution containing an active oxygen species remover, an active oxygen species remover The method of apply | coating or spraying the solution containing this to a foodstuff etc. is mentioned. The content ratio of the active oxygen species removing agent in the health food for removing active oxygen species is not particularly limited, but as a proportion of the plant extract based on the total amount of the health food for removing active oxygen species, 0.0001 to 10.0 % By weight is preferable, more preferably 0.0005 to 5.0% by weight, and particularly preferably 0.01 to 5% by weight.

  By ingesting the health food for removing reactive oxygen species of the present invention, it is possible to remove reactive oxygen species in the body and prevent inflammation, cancer, aging, lifestyle-related diseases and the like caused by the reactive oxygen species.

(5) Life miscellaneous goods The active oxygen species removing agent of the present invention can be used as life miscellaneous goods for removing active oxygen species by being included in life miscellaneous goods. Living miscellaneous goods containing the reactive oxygen species removing agent of the present invention are not particularly limited. For example, tableware, hardware, blades, thread, paper, film, toys, packaging materials, clothing, tools, stationery, home appliances , Musical instruments, furniture, curtains.
The method for containing the active oxygen species remover of the present invention in the household sundries is not particularly limited, but the method of mixing the active oxygen species remover with the household sundries material, the household sundries in the solution containing the active oxygen species remover or its Examples thereof include a method of immersing the material, and a method of applying or spraying a solution containing the active oxygen species removing agent to life miscellaneous goods or the material.

  By using the living oxygen for removing active oxygen species of the present invention, it is possible to remove active oxygen species in the living environment and prevent inflammation, cancer, aging, lifestyle-related diseases and the like caused by the active oxygen species.

(Example)
EXAMPLES The present invention will be specifically described below with reference to examples and comparative examples, but the present invention is not limited to these.

(Examples 1-3 and Comparative Examples 1-2) (Examination of radical scavenging ability by DPPH method)
19.8 mg of 2,2-diphenyl-1-picrylhydrazyl (α, α-diphenyl-β-picrylhydrazyl (DPPH) (manufactured by Sigma)) was dissolved in 50 ml of ethanol to prepare a DPPH solution having a concentration of 1 mM. 200 mg of dried plant powder of Sapindaceae Dodonaea viscosa was subdivided, immersed in 100 ml of ethanol at 25 ° C. for 1 week, and filtered to obtain a plant extract of 200 mge / ml (equivalent to milligrams of plant powder / ml). 998 μl of ethanol was added to 2 μl of the obtained plant extract with a concentration of 200 mge / ml to prepare a plant extract solution with a concentration of 0.4 mge / ml, and the plant extract solution with a concentration of 0.4 mge / ml was doubled with ethanol. Dilute to prepare a plant extract solution with a concentration of 0.2 mge / ml. A plant extract solution having a concentration of 0.4 mge / ml was diluted 4-fold with ethanol to prepare a plant extract solution having a concentration of 0.1 mge / ml.Vitamin C (manufactured by Sigma) was added to pure water. A vitamin C solution having a concentration of 10 μg / ml after dissolution was prepared in the proportions shown in Table 1. 1 mM DPPH solution, 0.4 mge / ml plant extract solution, 0.2 mge / ml concentration. A plant extract solution, a plant extract solution with a concentration of 0.1 mge / ml, a vitamin C solution with a concentration of 10 μg / ml, an acetic acid (sodium) buffer solution with a concentration of 10 mM, and ethanol are mixed, and 200 μge / ml, 100 μge / ml or Samples containing 50 μge / ml plant extract (Examples 1-3), samples not containing plant extract (Comparative Example 1), and samples containing 5 μg / ml vitamin C To prepare the reaction solution (Comparative Example 2).

  After reacting at room temperature for 10 minutes, the absorbance at 517 nm was measured, and the radical stability of Examples 1 to 3 and Comparative Example 2 was calculated with the absorbance of Comparative Example 1 being 100. The results are shown in Table 2.

(Examples 4 to 5 and Comparative Example 3) (Removal of intracellular reactive oxygen species)
Human leukemia-derived mononuclear cell line THP-1 was cultured using RPMI1640 supplemented with 10% fetal bovine serum as a medium until the required amount of cells was reached. Cells were seeded in ^6- well-plate (Corning) so that the cell density was 10 × 10 ⁶ cells / well (medium: 2 ml).
Thereafter, in Examples 4 and 5, 2 μl of a plant extract solution having a concentration of 200 mge / ml was added to the medium. In Comparative Example 3, 2 μl of dimethyl sulfoxide was added.
After culturing for 24 hours, the cells were collected and the cell density was adjusted to 3 × 10 ⁵ cells / ml to obtain a cell suspension. 1 ml of each cell suspension was transferred to a 1.5 ml capacity microtube. After washing the cells with 1 ml of phosphate buffer and centrifuging, the cells were resuspended in 1 ml of a fluorescent coloring reagent DCFH-DA (2 ′, 7′-dichlorofluorescein diacetate) (Sigma) with a concentration of 5 μM. It became cloudy and left at 37 ° C. for 15 minutes. After the coloring reagent was taken up into the cells, it was washed 3 times with 1 ml of phosphate buffer.
Thereafter, in Example 4, the cells were resuspended in 1 ml of a phosphate buffer containing a plant extract solution having a concentration of 200 μge / ml and cumene peroxide having a concentration of 100 μM. In Example 5 and Comparative Example 3, cells were resuspended in 1 ml of phosphate buffer containing cumene peroxide at a concentration of 100 μM.
Thereafter, each cell suspension was left at 37 ° C. for 1 hour. After the reaction, it was analyzed with a flow cytometer (BD FACSAria, manufactured by Becton-Dickinson), and the total fluorescence intensity in the cells was calculated. The results are shown in Table 3.

(Example 6 and Comparative Examples 4 to 5) (Removal of Nitric Oxide (NO))
The mouse mononuclear cell-derived cell line RAW264.7 was cultured using Dulbecco Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum until the required amount of cells was reached. The cells were seeded in 24 well-plate (Corning) so that the cell density was 2 × 10 ⁵ cells / well (medium: 1 ml) and cultured for 48 hours.
Thereafter, in Example 6, the medium was replaced with a medium containing a plant extract solution having a concentration of 200 μge / ml and 100 ng / ml lipopolysaccharide (LPS). In Comparative Example 5, the medium was changed to Dulbecco's Forked Modified Eagle Minimum Essential Medium (DMEM). In Comparative Example 6, the medium was replaced with a medium containing 100 ng / ml LPS.
After further culturing for 24 hours, NO contained in the culture supernatant was measured using a measurement kit Invitro NO assay kit (manufactured by Cell Biolabs), and the NO concentration (μM) was calculated. The results are shown in Table 4.

(Examples 7 to 8 and Comparative Examples 6 to 7) (SOD-like activity)
In Example 7, a plant extract solution having a concentration of 100 μge / ml, in Example 8, a plant extract solution having a concentration of 20 μge / ml, in Comparative Example 6 a vitamin C solution having a concentration of 5 μg / ml, and in Comparative Example 7 having a concentration of 50 μg / ml. Using the vitamin E solution, the SOD-like activity was calculated for each SOD activity using the SOD assay kit (manufactured by Cayman) according to the instruction manual of Cayman. The results are shown in Table 5.

(Examples 9 to 10 and Comparative Examples 8 to 9) (catalase-like activity)
In Example 9, a plant extract solution having a concentration of 100 μge / ml, in Example 10, a plant extract solution having a concentration of 20 μge / ml, in Comparative Example 8 a vitamin C solution having a concentration of 5 μg / ml, and in Comparative Example 9 having a concentration of 50 μg / ml. Catalase activity equivalent (U / ml) was calculated for each catalase-like activity using a vitamin E solution using Catalase assay kit (manufactured by Cayman) according to the instruction manual of Cayman. The results are shown in Table 6.

(Example 11 and Comparative Example 10) (Cytotoxicity, neutral red method)
Human leukemia-derived mononuclear cell line THP-1 was cultured using RPMI1640 supplemented with 10% fetal bovine serum as a medium until the required amount of cells was reached. Cells were seeded in 96 well-plate (Corning) so that the cell density was 5 × 10 ⁴ cells / well (medium: 1 ml).
In Example 11, after 24 hours, the medium was replaced with a medium containing a plant extract solution having a concentration of 200 μge / ml. In Comparative Example 10, after 24 hours, the medium was changed to a medium containing a DMSO solution having a concentration of 0.1%, and the medium was changed.
Further, after culturing for 24 hours, cytotoxicity was measured using a Neutral Red assay kit (manufactured by Sigma) according to the instruction manual of Sigma. The results are shown in Table 7.

(Example 12 and Comparative Example 11) (Cytotoxicity, MTT method)
Human leukemia-derived mononuclear cell line THP-1 was cultured using RPMI1640 supplemented with 10% fetal bovine serum as a medium until the required amount of cells was reached. Cells were seeded in 96 well-plate (Corning) so that the cell density was 5 × 10 ⁴ cells / well (medium: 1 ml).
In Example 11, after 24 hours, the medium was replaced with a medium containing a plant extract solution having a concentration of 200 μge / ml. In Comparative Example 10, after 24 hours, the medium was replaced with a medium containing a DMSO solution having a concentration of 0.1%.
After further culturing for 24 hours, cytotoxicity was measured using MTT assay kit (manufactured by Sigma) according to the instruction manual of Sigma. The results are shown in Table 8.

  The reactive oxygen species removing agent of the present invention, which comprises a plant extract of Sapindaceae Dodonaea viscosa as an active ingredient, is excellent in preventing inflammation, cancer, aging, lifestyle-related diseases, etc. caused by reactive oxygen species, has few side effects, and is a human body High safety. Therefore, this active oxygen species removing agent is contained and useful as a skin external preparation for removing active oxygen species, a health food for removing active oxygen species, and a daily miscellaneous product for removing active oxygen species.

Claims (5)

  A reactive oxygen species remover comprising, as an active ingredient, a plant extract of Sapindaceae Dodonaea viscosa.   The reactive oxygen species removing agent according to claim 1, wherein the reactive oxygen species is at least one selected from the group consisting of superoxide, singlet oxygen, hydroxy radical, hydroperoxy radical, and nitric oxide.   3. A skin external preparation for removing reactive oxygen species, comprising the reactive oxygen species removing agent according to claim 1 or 2.   A health food for removing active oxygen species, comprising the active oxygen species removing agent according to claim 1.   A household oxygen for removing active oxygen species comprising the active oxygen species removing agent according to claim 1.