Antioxidants for skin, antioxidant foods or beverages for skin, antioxidant pharmaceuticals for skin, and antioxidant cosmetics for skin.

Abstract

The present invention provides an anti-aging agent for the skin that has an anti-aging effect on the skin. [Solution] The solution comprises nicotinamide mononucleotide, docosahexaenoic acid, and eicosapentaenoic acid.

Description

【Technical Field】 【0001】 The present invention relates to anti- Oxidizing agent, skin foods or anti- oxidation skin beverages, or anti- oxidation skin pharmaceuticals. oxidation and Anti- Oxidative cosmetics skin 【Background Art】 【0002】 As the three major aging reactions of the living body, glycation reaction, oxidation reaction, and chronic inflammation (which can also be said to be vascular aging such as arteriosclerosis) are mentioned. In addition, as the three major causes of aging, obesity, decreased efficiency of immune function, and accumulation of senescent cells are mentioned. Reactive oxygen species accumulate in senescent cells, and the accumulation of reactive oxygen species further accelerates cell aging, so its elimination function is highly regarded. 【0003】 In recent years, nicotinamide mononucleotide (hereinafter sometimes referred to as NMN) has attracted attention. NMN is a derivative of vitamin B3 (niacin) and is converted into a molecule called nicotinamide adenine dinucleotide (oxidized form) (NAD + ) in the body. Since NAD + activates the sirtuin gene, which is also called an anti-aging gene or a longevity gene, an anti-aging effect is expected. Studies have also been conducted on combining NMN with other substances. For example, in Patent Document 1, a hepatocyte activator has been proposed by combining NMN with other substances including a natural-derived calcined zeolite powder and a reducing agent for food additives. 【Prior Art Documents】 【Patent Documents】 【0004】 【Patent Document 1】 Japanese Patent No. 7610823 【Summary of the Invention】 【Problems to be Solved by the Invention】 【0005】 While Patent Document 1 concerns hepatocytes, there is a need for a method that is effective against other cells and tissues. 【0006】 The objective of the present invention is to provide resistance to the skin. oxidation Skin anti- oxidation Agent, this skin anti oxidation Using the agent skin Targeting resistance oxidation Antimicrobial agents for food or skin use oxidation beverages, anti- oxidation Pharmaceuticals for use and anti-dermatological applications oxidation Provides cosmetics for use thing That is the case. [Means for solving the problem] 【0007】 The inventors of the present invention discovered that the above objective can be achieved by combining NMN with a predetermined substance, and thus completed the present invention. 【0008】 In other words, according to the present invention, (1) A skin anti-aging agent comprising nicotinamide mononucleotide, docosahexaenoic acid and eicosapentaenoic acid, (2) The anti-aging agent for skin according to (1), further comprising 5-aminolevulinic acid, (3) A skin anti-aging agent according to (1) or (2), comprising 80 to 120 parts by weight of eicosapentaenoic acid per 100 parts by weight of docosahexaenoic acid. (4) The anti-aging agent for skin according to (3), comprising 100 parts by weight of eicosapentaenoic acid per 100 parts by weight of docosahexaenoic acid, (5) An anti-wrinkle agent comprising the anti-aging agent for skin described in (1) or (2), (6) A food or beverage for skin use that contains the anti-aging agent for skin described in (1) or (2), (7) A dermatological anti-aging drug comprising the dermatological anti-aging agent described in (1) or (2), (8) A cosmetic for skin that contains the anti-aging agent for skin described in (1) or (2), is provided. 【Effects of the Invention】 【0009】 According to the present invention, there are provided an anti-aging agent for skin, which has an anti-aging effect on the skin, i.e., the dermis, an anti-aging food or an anti-aging beverage for skin, an anti-aging pharmaceutical, and an anti-aging cosmetic for skin. oxidation using this anti-aging agent for skin anti-aging food or anti-aging beverage for skin oxidation using this anti-aging agent for oxidation anti-aging food or anti-aging beverage for skin oxidation using this anti-aging agent for oxidation anti-aging pharmaceutical, and an anti-aging cosmetic for skin are provided. ru. 【Brief Description of the Drawings】 【0010】 [Figure 1] It is a graph showing the measurement results of ORAC in the examples. [Figure 2] It is a graph showing the measurement results of Superoxide (SOD) against hydrogen peroxide in the examples. [Figure 3] It is a graph showing the results of examination on the anti-glycation effect in the examples. [Figure 4] It is a graph showing the results of examination on the anti-wrinkle and moisturizing effects in the examples. 【Modes for Carrying Out the Invention】 【0011】 Hereinafter, the anti-aging agent for skin of the present invention and its production method will be described. 【0012】 The anti-aging agent for skin of the present invention comprises nicotinamide mononucleotide, docosahexaenoic acid (hereinafter sometimes referred to as "DHA") and eicosapentaenoic acid (hereinafter sometimes referred to as "EPA"). 【0013】 (NMN) NMN has α-type and β-type optical isomers, and either the α-type or the β-type may be used, but it is preferable to use the β-type. Furthermore, NMN may be derived from natural products or artificially synthesized. If it is derived from natural products, it is preferable to perform extraction using known extraction methods. 【0014】 (DHA) The DHA used in this invention is a type of omega-3 fatty acid, which is mainly found in abundance in marine foods. 【0015】 DHA can be obtained from marine fish oil and algae. For fish oil, it is particularly preferable to use fish oil from oily fish such as sardines, mackerel, and saury. Specifically, it can be obtained by extracting from fish oil or algae using pressing or solvent extraction methods, followed by purification and concentration. Furthermore, in order to obtain a high concentration of DHA, the manufacturing process may include steps such as concentration by molecular distillation or esterification followed by further separation and concentration. 【0016】 (EPA) EPA is a type of omega-3 fatty acid, particularly abundant in marine foods. EPA can be obtained from marine fish oil and algae. For fish oil, it is preferable to use fish oil from oily fish such as sardines, mackerel, and saury. Specifically, EPA can be obtained from fish oil or algae by pressing or solvent extraction, followed by purification and concentration. Furthermore, in order to obtain high concentrations of EPA, the manufacturing process may include steps such as concentration by molecular distillation or esterification followed by further separation and concentration. 【0017】 For the DHA and EPA used in the present invention, LDR-DHA / EPA Premier® is preferred, which contains EPA in an amount of 80 to 120 parts by weight, more preferably 90 to 110 parts by weight, and particularly preferably 100 parts by weight, relative to 100 parts by weight of DHA. In LDR-DHA / EPA Premier®, the ratio of DHA to EPA is approximately equal, which is a unique ratio not seen elsewhere. In LDR-DHA / EPA Premier, DHA and EPA are preferably contained in amounts of 10 to 50% by weight, more preferably 20 to 40% by weight, and even more preferably 25 to 35% by weight, respectively. 【0018】 (5-ALA) The anti-aging agent for skin of the present invention may further contain 5-aminolevulinic acid (hereinafter sometimes referred to as "5-ALA") in addition to the above-mentioned NMN, DHA, and EPA. 5-ALA may be produced by chemical synthesis or by microbial fermentation. 【0019】 (Anti-aging agent for skin) The anti-aging agent for skin of the present invention comprises NMN, DHA, and EPA, and optionally 5-ALA. The mixing ratio is not particularly limited, but it is preferable to mix 1 to 10,000 parts by weight of DHA per 100 parts by weight of NMN. It is also preferable to mix 1 to 10,000 parts by weight of EPA per 100 parts by weight of NMN. Furthermore, when 5-ALA is included, it is preferable to mix 1 to 10,000 parts by weight of 5-ALA per 100 parts by weight of NMN. The above components may be mixed by known methods. As mentioned above, it is preferable to use LDR-DHA·EPA Premier®, which contains approximately equal amounts of DHA and EPA. 【0020】 The anti-aging agent for skin of the present invention may contain other ingredients, provided they do not inhibit the effects of the present invention. Other ingredients include resveratrol, L-cystine, vitamin C, pineapple extract (containing ceramide), red orange extract, pasenol, GABA, coenzyme Q10, lactic acid bacteria powder, odorless garlic powder, alpha-lipoic acid, ginger powder, vitamin A, vitamin D, vitamin E, biotin, vitamin B1, vitamin B2, vitamin B6, and vitamin B1. 12 Examples include niacin, pantothenic acid, folic acid, calcium, magnesium, iron, manganese, copper, selenium, chromium, molybdenum, zinc, valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, histidine, glycine, arginine, glutamic acid, alanine, aspartic acid, proline, serine, and tyrosine. 【0021】 The anti-aging agent for skin of the present invention exhibits excellent antioxidant effects in the skin. Furthermore, it also exhibits excellent anti-glycation, anti-wrinkle, and moisturizing effects in the skin. In other words, it demonstrates superior anti-aging effects on the skin. 【0022】 (Anti-aging foods or beverages for the skin) The anti-aging agent for skin of the present invention can be incorporated into anti-aging foods or beverages for skin. Examples of anti-aging foods for skin include bread, noodles, confectionery, processed meat products, processed seafood products, frozen foods, jellies, ice cream, dairy products, and various seasonings. In addition to general foods, it can also be incorporated into foods for specified health uses, quasi-drugs, health foods, and supplements. Examples of anti-aging beverages for skin include soft drinks, dairy beverages, alcoholic beverages, tea, black tea beverages, coffee, fruit juices, carbonated drinks, mineral water, and fruit and vegetable beverages. 【0023】 Furthermore, when manufacturing foods or beverages containing the anti-aging agent for skin of the present invention, additives such as sweeteners, colorants, preservatives, thickeners, stabilizers, gelling agents, antioxidants, color fixatives, bleaching agents, emulsifiers, leavening agents, acidulants, glazing agents, and flavorings; solvents; and oils may be added as needed, to the extent that they do not interfere with the effects of the present invention. These additives may be used individually or in combination of two or more types. 【0024】 The proportion of the anti-aging agent for skin of the present invention incorporated into the above-mentioned anti-aging food or beverage for skin can be appropriately adjusted depending on the intended use, but the proportion of the ingredient formulation incorporated into the above-mentioned food or beverage is preferably 0.01 to 20% by weight, more preferably 0.01 to 15% by weight, and even more preferably 0.1 to 10% by weight. 【0025】 Furthermore, the anti-aging food or beverage for skin of the present invention may be in the same form as the oral formulations such as tablets, capsules, and syrups used in the anti-aging pharmaceuticals for skin described later. 【0026】 (Anti-aging drugs for skin) Furthermore, the anti-aging agent for skin of the present invention can be incorporated into anti-aging pharmaceuticals for skin. Examples of anti-aging pharmaceuticals for skin include preventive drugs and therapeutic drugs. 【0027】 When incorporated into anti-aging drugs for skin use, the active ingredient may be used alone or mixed with generally pharmaceutically acceptable excipients to form a formulation. Regarding the form of administration, there are no particular restrictions, and the appropriate form can be selected as appropriate depending on the purpose of treatment or prevention. Examples of administration methods include oral formulations such as tablets, granules, capsules, pills, powders, liquids, suspensions, emulsions, syrups, elixirs, and extracts, or parenteral formulations such as injections, liquids, suppositories, ointments, patches, poultices, and lotions. 【0028】 Furthermore, in the case of tablets, granules, pills, capsules, and powders, additives such as excipients, binders, disintegrants, and lubricants may be included. Examples of excipients include starch, carboxymethylcellulose, sucrose, dextrin, and corn starch. 【0029】 Examples of binders include crystalline cellulose, crystalline cellulose-carmellose sodium, methylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, carmellose sodium, ethylcellulose, carboxymethyl ethylcellulose, hydroxyethylcellulose, wheat starch, rice starch, corn starch, potato starch, dextrin, pregelatinized starch, partially pregelatinized starch, hydroxypropyl starch, pullulan, polyvinylpyrrolidone, aminoalkyl methacrylate copolymer E, aminoalkyl methacrylate copolymer RS, methacrylic acid copolymer L, methacrylic acid copolymer, polyvinyl acetal diethylaminoacetate, polyvinyl alcohol, gum arabic, gum arabic powder, agar, gelatin, white shellac, tragacanth, refined sucrose, and macrogol. 【0030】 Examples of disintegrants include crystalline cellulose, methylcellulose, low-substituted hydroxypropylcellulose, carmellose, carmellose calcium, carmellose sodium, croscarmellose sodium, wheat starch, rice starch, corn starch, potato starch, partially pregelatinized starch, hydroxypropyl starch, carboxymethyl starch sodium, and tragacanth. 【0031】 Examples of lubricants include wheat starch, rice starch, corn starch, stearic acid, calcium stearate, magnesium stearate, hydrated silicon dioxide, light anhydrous silicic acid, synthetic aluminum silicate, dried aluminum hydroxide gel, talc, magnesium aluminometasilicate, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, sucrose fatty acid esters, waxes, hydrogenated vegetable oils, and polyethylene glycol. 【0032】 Furthermore, in the case of liquid preparations, syrups, suspensions, emulsions, and elixirs, in addition to commonly used inert diluents such as water and vegetable oil, colorants, flavoring agents, and fragrances may be included as additives. 【0033】 Furthermore, in the case of injectable preparations, additives such as suspensions, emulsions, and solvents for use may be included. In the case of ointments and suppositories, additives such as fats, fatty oils, lanolin, petrolatum, paraffin, waxes, resins, plastics, bases, glycols, higher alcohols, water, emulsifiers, and suspending agents may be included. In the case of poultices, additives such as glycerin, water, water-soluble polymers, and superabsorbent polymers may be included. In the case of lotions, additives such as solvents, emulsifiers, and suspending agents may be included. 【0034】 Furthermore, the anti-aging agent for skin of the present invention can also be incorporated into cosmetics. Examples of cosmetics include lotions, emulsions, facial washes, cleansers, serums, creams, foundations, eyebrow products, mascaras, eyeshadows, eyeliners, lipsticks, lip glosses, blushes, face powders, and nail polishes. The cosmetic can also be in the form of a liquid, cream, solid, stick, or powder. [Examples] 【0035】 The present invention will be described below with reference to examples, but the present invention is not limited thereto. In these examples, parts and percentages are based on weight unless otherwise specified. 【0036】 (Example 1: Measurement of ORAC) The antioxidant capacity of the samples was measured using the ORAC (Oxygen Radical Absorbance Capacity) method with Trolox as the standard antioxidant. The measurements were performed using the ORAC Activity Assay Kit. The samples are as follows. The samples were prepared so that the main component was present in concentrations of several tens of μg / mL, and serially diluted. 【0037】 Sample 1: DHA and EPA (combined in a 1:1 weight ratio) Sample 2: NMN Sample 3: 5-ALA Sample 4: DHA, EPA, NMN, and 5-ALA (DHA and EPA are in a weight ratio of 1:1) Sample 5: DHA, EPA, and NMN (DHA and EPA are combined in a weight ratio of 1:1) 【0038】 The results are shown in Figure 1. The values ​​on the horizontal axis of "ORAC" in Figure 1 indicate the dilution ratio. As shown in Figure 1, sample 4 showed a high ORAC value. 【0039】 (Example 2: Measurement of Superoxide (SOD) in relation to hydrogen peroxide) Human skin fibroblasts were incubated in a culture medium supplemented with 0.2 mM hydrogen peroxide for 2 hours, then washed and removed with PBS, and transferred to a normal medium for continued culture. Materials 1-5 above were added after the addition of hydrogen peroxide, and co-cultured for 4 hours. Then, the cells or the culture medium were collected. 【0040】 Subsequently, the measurement of Superoxide (SOD) was performed using an SOD Assay Kit (Cayman Chemical, Ann Arbor, MI, USA). After culturing the cells that had undergone various treatments for 24 hours, lysis buffer was added, homogenized, and centrifuged. Then, 10 μL of each sample was mixed with 200 μL of xanthine oxidase at room temperature. After 30 minutes, the absorbance at 450 nm was measured. 【0041】 The results are shown in Figure 2. In Figures 2 and 3, the numerical values on the horizontal axis indicate the number of times of serial dilution. 1 / 5 < n indicates that 5-fold dilution was performed n times (n is an integer from 1 to 7), and "1" indicates the sample before serial dilution. 【0042】 As shown in Figure 2, in Samples 4 and 5, an SOD removal or inhibitory effect was observed in skin fibroblasts. 【0043】 (Example 3: Examination of anti-glycation effect) (Preparation of AGE) Bovine serum-derived albumin (25 mg / mL) was dissolved in DL-glyceraldehyde to make it 0.1 M, and AGE was obtained by culturing in phosphate-buffered saline (pH 7.4; sometimes referred to as "PBS" in this example) at 37°C for 1 week. Human skin fibroblasts (Normal Human Dermal Fibroblasts (NHDF), juvenile foreskin (C-12300, PromoCell)) were cultured in a dedicated medium, Fibroblast Growth Medium (C-23010, PromoCell) in a 37°C, 5% CO2 incubator. Next, human dermal fibroblasts were treated with AGE (100 μg / mL), and samples 1-5, each serially diluted (1x, 1 / 10x, 1 / 100x, and 1 / 1000x), were added to each sample and held at 37°C for 4 hours. 【0044】 Then, total RNA was extracted from the cells using Trizol reagent (Ambion), and mRNA levels of AGE receptors (CD-36, AGE-R1, and AGE-R3) were measured by qRT-PCR (one-step quantitative reverse transcription-polymerase chain reaction). The results are shown in Figure 3. CD-36, AGE-R1, and AGE-R3 are AGE receptors involved in the degradation and removal of AGEs. Furthermore, the qRT-PCR method will be described in detail later in the section titled "qRT-PCR Method". As shown in Figure 3, elevated mRNA levels of CD-36, AGE-R1, and AGE-R3 were observed in AGE-induced glycation stress cells at high concentrations of DHA & EPA (Sample 1). Furthermore, all samples showed a tendency to increase CD-36 and AGE-R1 levels. 【0045】 (Example 4: Examination of anti-wrinkle and moisturizing effects) Human dermal fibroblasts were incubated for 2 hours in a culture medium supplemented with 0.2 mM hydrogen peroxide, then washed with PBS, and cultured in standard medium. Substances 1-4 were added after the addition of hydrogen peroxide, and co-cultured for 4 hours. After that, the cells or culture medium were collected. Total RNA was extracted from the cells using Trizol reagent (Ambion), and the mRNA level of hyaluronic acid was measured by qRT-PCR. The results are shown in Figure 4(a). 【0046】 Furthermore, using a predetermined culture medium, samples 1 to 4, each serially diluted (1x, 1 / 10x, 1 / 100x, and 1 / 1000x), were added to stem cells (human mesenchymal stem cells) and cultured in an incubator at 37°C. Exosomes were then extracted from the supernatant of the culture medium containing the stem cells using an ultracentrifuge. The extracted exosomes were added to dermal fibroblasts and incubated for 4 hours. 【0047】 The dermal fibroblasts used were human dermal fibroblasts (NHDF), juvenile foreskin (C-12300, PromoCell), cultured in a dedicated medium, Fibroblast Growth Medium (C-23010, PromoCell), at 37°C in a 5% CO2 incubator. 【0048】 Then, total RNA was extracted from the cells using Trizol reagent (Ambion), and the mRNA levels of collagen, elastin, and hyaluronic acid were measured by qRT-PCR. The results for collagen are shown in Figure 4(b), for elastin in Figure 4(c), and for hyaluronic acid in Figure 4(d). 【0049】 As shown in Figure 4, under H2O2 oxidative stress, an increase in hyaluronic acid synthase mRNA levels was observed in sample 1 (DHA & EPA). Furthermore, when exosomes obtained by stimulating stem cells with each sample were added to skin fibroblasts, the mRNA levels of collagen and elastin increased. In addition, hyaluronic acid synthase mRNA levels increased in all samples. This is thought to be due to the secretion of exosomes different from those normally secreted. In other words, these results suggest anti-wrinkle and moisturizing effects in skin cells. 【0050】 (qRT-PCR method) The qRT-PCR method used for measuring AGE receptors, collagen, elastin, and hyaluronic acid, as described above, is explained. In the qRT-PCR method, 500 ng of obtained total RNA was used as a PCR template, and a one-step RT-PCR was performed using the Luna Universal One-Step qRT-PCR Kit, which allows for cDNA synthesis by reverse transcription (RT reaction) and quantitative PCR to be performed in a single test tube. 【0051】 A Takara Thermal Cycler Dice Real Time System II PCR instrument was used. For one reaction system, 10 μL of Luna Universal One-Step Reaction Mix (2x), 1 μL of Luna WarmStart RT Enzyme Mix (20x), 0.8 μL of Forward primer (10 μM), 0.8 μL of Reverse primer, and Total RNA as the PCR template were prepared by adding 20 μL of Nuclease-free Water. The reaction was carried out under conditions of 30 seconds per cycle at 95°C, followed by 50 cycles of 5 seconds at 95°C and 30 seconds at 60°C.

Claims

[Claim 1] A skin antioxidant comprising nicotinamide mononucleotide, docosahexaenoic acid, eicosapentaenoic acid, and 5-aminolevulinic acid. [Claim 2] The skin antioxidant according to claim 1, comprising 80 to 120 parts by weight of eicosapentaenoic acid per 100 parts by weight of docosahexaenoic acid. [Claim 3] The skin antioxidant according to claim 2, comprising 100 parts by weight of eicosapentaenoic acid per 100 parts by weight of docosahexaenoic acid. [Claim 4] A skin antioxidant food or skin antioxidant beverage comprising the skin antioxidant described in claim 1 or 2. [Claim 5] A skin antioxidant pharmaceutical comprising the skin antioxidant described in claim 1 or 2. [Claim 6] A skin antioxidant cosmetic comprising the skin antioxidant described in claim 1 or 2.

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