WO2017213346A1 - Composition containing diosmin or salt thereof as active ingredient and having skin moisturizing improvement, skin exfoliation, skin elasticity enhancement, erythema inhibition, skin wrinkle alleviation, or skin photoaging retardation effect - Google Patents

Abstract

The present invention relates to a cosmetic composition, a health functional food composition, and medicine and quasi-drug compositions, containing diosmin or a salt thereof as an active ingredient and having skin wrinkle alleviation, skin moisturizing improvement, skin exfoliation, skin elasticity enhancement, skin wrinkle alleviation, erythema inhibition, and skin photoaging retardation effects. Diosmin or a salt thereof according to the present invention exhibits skin moisture content improvement, skin moisture loss reduction, procollagen secretion increase, collagen biosynthesis promotion, collagen fiber damage inhibition, collagen fiber decomposition inhibition, erythema inhibition, and epidermal hypertrophy inhibition activities, and thus can be usefully applied as a material for a functional cosmetic, a health functional food, a medicine, a quasi drug, or the like for providing skin moisturizing improvement, skin exfoliation, skin elasticity enhancement, skin wrinkle alleviation, erythema inhibition, skin wrinkle alleviation, and/or skin photoaging retardation effects.

Description

A composition having the effect of improving skin moisturizing, removing dead skin cells, enhancing skin elasticity, inhibiting erythema, improving skin wrinkles or improving skin photoaging, containing diosmin or a salt thereof as an active ingredient.

This application claims the priority of Korean Patent Application No. 10-2016-0071737, filed June 09, 2016, the entirety of which is a reference of the present application.

The present invention relates to a composition containing diosmin or a salt thereof as an active ingredient, and more particularly, to enhance skin moisture content, decrease skin moisture evaporation amount, increase procollagen secretion amount, promote collagen biosynthesis, inhibit collagen fiber damage, collagen fiber degradation Cosmetic composition having the effect of improving skin moisturizing, removing dead skin cells, enhancing skin elasticity, inhibiting erythema, improving skin wrinkles and / or improving skin photoaging by using diosmin or a salt thereof that inhibits, inhibits erythema and thickening of the skin epidermal layer. , Dietary supplements, pharmaceuticals and quasi-drug compositions can be provided.

Recently, as life expectancy is extended due to the development of medical technology and the quality of life and the desire for healthy and beautiful life are increasing, interest in skin beauty and health is expanding. Accordingly, various cosmetic functional cosmetics have been developed according to the purpose of maintaining healthy skin, and research for preventing, alleviating and improving skin wrinkle formation is being actively conducted. In addition, cosmetic ingredients have recently reached the skin dermis to supply nutrients, and as a result of the change in the perception that skin nutrients or functional ingredients can be ingested to improve skin beauty, resulting in inner beauty foods Excavations are also actively underway.

The skin is composed of three layers: epidermis, dermis, and hypodermis. The epidermis, especially the stratum corneum, the outermost layer of the epidermis, acts as a barrier to the skin to lose water and electrolytes. On the other hand, the dermal layer plays a role in maintaining the elasticity of the skin and supporting the structure through collagen and elastin synthesis. In other words, collagen and elastin are major proteins produced in fibroblasts and are involved in skin mechanical firmness, tissue binding strength and elasticity. Collagen forms various isoforms according to its shape and structural features, and there are a total of 28 collagen isotypes in human tissues, of which collagen is present in Type 1, 3. 4. 6. 13, 14, 17 and the like are known. Collagen types 1 and 3 make up the interstitial components of the dermal layer, and collagen type 7 is the major component of the dermal and epidermal junctions.

Type I collagen is the largest amount of extracellular matrix protein in the skin connective tissue, and there are other proteins such as elastin, fibronectin, integrin, fibrillin, and proteoglycan. The newly synthesized procollagen is secreted into the extracellular space of skin cells through an enzymatic reaction to form a microfibril of triple helix configuration, and the microfibrils are leucine-rich small proteoglycans (leucine). It combines with rich small proteoglycans to form fibrils. As a result, the fibrils thus formed form collagen fibers that provide skin binding and elasticity.

Skin aging is known to decrease collagen content, a protein that accounts for most of the collagen of skin dermis. Collagen decreases the skin's tension and strength. have. Skin aging is largely divided into endogenous aging due to physiological aging and photoaging caused by continuous ultraviolet radiation (UV) exposure. Repeated ultraviolet exposure results in increased collagen degrading enzymes and causing denaturation and destruction of collagen fibers, reducing the elasticity of the skin and promoting the production of wrinkles. That is, the generation of reactive oxygen species (ROS) is increased in skin tissues exposed to ultraviolet rays continuously, and the latter has a signaling system mediated by growth factor receptor (EGF-R), tumor necrosis factor (TNF) -receptor, etc. It promotes proinflammatory cytokine production. Activation of these receptors consists of continuous phosphorylation that activates downstream signaling mediators, including mitogen-activated protein kinase (MAPK), resulting in activator protein-1 (AP-1). And activating transcription factors such as nuclear factor κB (NF-κB) to induce an inflammatory response and to promote the activity of collagenase such as matrix metalloproteinases (MMPs) to reduce skin elasticity and promote wrinkle formation do. AP-1 regulates the expression of numerous genes involved in cell growth and differentiation and strongly regulates the expression of some MMPs. Among the MMPs whose expression is regulated by AP-1, MMP-1 is known as collagenase 1, and is based on type 1 and 3 collagen.

On the other hand, diosmin is the main component of the drug that is commercially available for the treatment of diseases such as chronic venous insufficiency, hemorrhoids, lymphedema. It is generally recommended to take 500 mg diosmin tablets twice a day for the treatment of these diseases.In acute illnesses, 1,000 mg three times a day and 1,000 mg twice daily for three days. After that, it is recommended to take 500 mg twice a day for two months.

In a multinational clinical trial conducted over two years in 23 countries, 5,052 patients with chronic venous insufficiency had been treated with 450 mg diocimine daily for six months and found improvement. In addition, clinical trials in which patients with severe venous insufficiency with venous ulceration were difficult to treat were given 900 mg diosmin in addition to standard treatment protocols (cleaning, compression therapy, skin care in adjacent areas). In the study, only 28% of patients who received standard treatment protocols were completely cured of ulcers, while 47% of patients with diosmin were fully cured.

Several large multinational clinical studies have reported the effects of diosmin in relieving acute and chronic hemorrhoids. In a double-blind, placebo-controlled study of 120 patients with hemorrhoids, pain, bleeding, swelling and pruritus after taking 500 mg of a flavonoid mix of 90% diosmin and 10% hesperidin for two months a day It was confirmed that this was alleviated. Diosmin is also very effective in relieving hemorrhoidal symptoms associated with pregnancy and has no negative effect on the maintenance of pregnancy, fetal development, birth weight, neonatal growth rate and intake, and does not mutate or affect reproductive function. Turned out to be.

Diosmin has an effect on the lymphatic system to increase the lymph flow (lymph flow). In addition to traditional treatments, the use of diosmin for the treatment of upper limb lymphedema has been shown to improve symptoms. Diosmin also showed significant symptomatic relief in lymphedema animal models such as burns and lung cotusions.

Diosmin has been shown to be effective in premenstrual syndrome, viral infections, mastopathy, colitis, etc. In addition, diosmin has been shown to have anti-cancer effects and cell proliferation inhibitory effects in various animal and cell line models. . However, there is no report on the anti-wrinkle or collagen promoting activity of diosmin.

The present inventors conducted a study to develop a food or cosmetic material that is effective in improving wrinkles by inhibiting the action of collagen degrading enzymes in natural products with fewer side effects and promoting collagen synthesis, and thus, diosmin improves skin moisturization. The present invention was completed by confirming that exfoliation, skin elasticity enhancement, erythema suppression, skin wrinkle improvement and / or skin photoaging improvement were shown.

An object of the present invention is to provide a cosmetic composition having an effect of improving skin moisturizing, skin exfoliation, skin elasticity enhancement, erythema suppression, skin wrinkle improvement and / or skin photoaging.

Another object of the present invention is to provide a health functional food composition having an effect of improving skin moisturizing, removing skin dead skin, enhancing skin elasticity, suppressing erythema, improving skin wrinkles and / or improving skin photoaging.

Still another object of the present invention is to provide a pharmaceutical or quasi-drug composition having the effect of improving skin moisturizing, removing skin dead skin, enhancing skin elasticity, suppressing erythema, improving skin wrinkles and / or improving skin photoaging.

In addition, the present invention provides a skin moisturizing improvement, skin exfoliation, skin elasticity enhancement, erythema suppression, wrinkle improvement and / or skin comprising administering a composition containing diosmin or a pharmaceutically acceptable salt thereof as an active ingredient It is intended to provide a method for improving photoaging.

Furthermore, the present invention comprises diosmin or a pharmaceutically acceptable salt thereof as an active ingredient for improving skin moisturizing, skin exfoliation, skin elasticity, erythema suppression, skin wrinkle improvement and / or preparation of skin photoaging improving agent. It is intended to provide a use of the composition.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

The present invention provides a cosmetic composition having a skin moisturizing improvement, skin exfoliation, skin elasticity enhancement, erythema suppression, skin wrinkle improvement and / or skin photoaging improvement effect including diosmin or a cosmetically acceptable salt thereof as an active ingredient. to provide.

The composition containing the diosmin of the present invention or a salt thereof as an active ingredient may improve skin moisture content, reduce skin moisture evaporation amount, increase collagen secretion amount, promote collagen biosynthesis, inhibit collagen fiber damage, inhibit collagen fiber degradation, inhibit erythema and skin epidermal layer. It has excellent thickening inhibitory activity, so it can be usefully used to improve skin moisturizing, skin exfoliation, skin elasticity improvement, erythema suppression, skin wrinkle improvement and / or skin photoaging.

In addition, since the diosmin of the present invention is a natural product, it is safe for the human body and has less side effects, and thus may be used in various materials such as cosmetics, health foods, medicines or quasi-drugs.

1 is a graph showing the results of measuring procollagen secretion by ELISA assay after irradiating UVB at 20 mJ / cm 2 to human dermal fibroblasts for 24 hours with 100 μM of vitamin C or diosmin (-UVB: Normal group without UVB treatment, + UVB: UVB treated control group, DM: UVB treated group, diosmin treated group, VitC: UVB treated group, vitamin C treated group, DM + VitC: UVB treated group Then treated with diosmin and vitamin C).

Figure 2 is a graph showing the weight gain and dietary intake of mice fed the experimental diet having the composition of Table 1 (A: weight, B: weight gain, C and D: diet intake).

Figure 3 is a graph showing the water content (A), water evaporation (B), elasticity (C) and erythema index (D) of the skin tissue of mice fed the experimental diet having the composition of Table 1.

Figure 4 is a photograph showing the back skin tissue of each mouse fed the experimental diet having the composition of Table 1.

5 is a photograph (A) and a graph (B) showing the degree of wrinkle formation of the back skin tissue of the mouse fed the experimental diet having the composition of Table 1.

6 is a graph (A) and skin epidermal layer photograph (B) of measuring skin thickness of mice fed an experimental diet having the composition of Table 1. FIG.

Figure 7 is a photograph observing the change in the amount and morphology of collagen fibers in mice fed an experimental diet having the composition of Table 1.

FIG. 8 shows the results of RT-PCR measurement of expression changes of collagen and MMP genes in hairy mouse back tissue treated with diosmin.

Hereinafter, the present invention will be described in more detail.

As described above, diosmin has been known to be effective in treating diseases such as chronic venous insufficiency, hemorrhoids and lymphedema, premenstrual syndrome, viral infections, breast pain, colitis, etc. There has been no report on moisturizing, skin exfoliation, skin elasticity improvement, erythema suppression, skin wrinkle improvement and / or skin photoaging improvement.

The present inventors confirmed the novel activity of diosmin related to skin health, and developed a cosmetic composition, a dietary supplement, a pharmaceutical and a quasi-drug composition containing diosmin or a salt thereof as an active ingredient.

The composition provided by the present invention has excellent skin hydration content, reduced skin moisture evaporation amount, increased procollagen secretion, increased collagen biosynthesis, inhibited collagen fiber damage, inhibited collagen fiber degradation, erythema suppression and thickening inhibitory effect of skin epidermal layer. It can be usefully used for improvement, skin moisturizing improvement, skin exfoliation, skin elasticity improvement, erythema suppression, skin wrinkle improvement and / or skin photoaging improvement.

Therefore, the present invention is a cosmetic composition having a skin moisturizing improvement, skin exfoliation, skin elasticity enhancement, erythema suppression, wrinkle improvement and / or skin photoaging improvement effect containing diosmin or a cosmetically acceptable salt thereof as an active ingredient To provide.

Diosmin is a flavonoid glycoside (flavone) -based compound represented by Formula 1 below, and the name designated by the International Pure and Applied Chemistry Organization (IUPAC) is 5-hydroxy-2- (3-hydroxy-4 -Methoxyphenyl) -7-[(2S, 3R, 4S, 5S, 6R) -3,4,5-trihydroxy-6-[[(2R, 3R, 4R, 5R, 6S) -3,4 , 5-trihydroxy-6-methyloxan-2-yl] oxymethyl] oxan-2-yl] oxychromen-4-one [5-Hydroxy-2- (3-hydroxy-4-methoxyphenyl) -7 -[(2S, 3R, 4S, 5S, 6R) -3,4,5-trihydroxy6-[[(2R, 3R, 4R, 5R, 6S) -3,4,5-trihydroxy-6-methyloxan-2- yl] oxymethyl] oxan-2-yl] oxychromen-4-one], also called diosmetin. The CAS registry number for diosmin is 520-27-4, the structural formula is C28H32O15, and the molecular weight is 608.5 g / mol.

[Formula 1]

The appearance of the diosmin compound is a pale yellow or grayish yellow powder. Diosmin is well soluble in water and in small amounts in dimethyl sulfoxide (50 mg / ml at 25 ° C) and ethanol (<1 mg / ml at 25 ° C). The melting point of diosmin is 274 ° C, the boiling point is 926.8 ° C (at 760 mmHg), the density is 1.7 g / cm3, the refractive index is n20D 1.71, and the pKa value is: 6.1.

Diosmin is a flavonoid glycoside material present in various plants and can be extracted from plants or synthesized from flavonoid hesperidin. Among the citrus fruits, the particularly unripe Meyer lemon (3.06 g diosmin / 100 g of dry weight) and Buddha's finger fruits (3.64 g / 100 g of dry weight) contain the highest amount of diosmin Known as In addition, Vetches (Asian and Caucasian vetch species) native to North America, Europe, Africa, and Asia are representative plants containing a large amount of diosmin, and the leaves contain about 2% of diosmin. Herb hyssop (also called Hyssopus officinalis or Hyssopus decumbens), native to southern and central Asia, is also a source of diosmin, especially in leaves and flowers.

The method for obtaining the diosmin of the present invention is not particularly limited, and may be isolated from natural products, chemically synthesized using a known production method, or commercially available.

Although the LD ₅₀ value of diosmin is not known much, it has been reported that it is safe to administer more than 3000 mg / kg when administered orally to mice. Diosmin has also been reported to be very limited in migration through the placenta and into breast milk.

Therefore, since diosmin is administered at 100 mg / kg to 3,000 mg / kg, preferably 500 mg / kg to 2,000 mg / kg, the toxicity is very low or not, so cosmetic compositions, dietary supplement compositions, pharmaceutical or quasi-drug compositions, etc. Can be used as

In the present invention, the diosmin may include a diosmin hydrate, a diosmin derivative, and the like within a range having the same efficacy, and may include a solvate or stereoisomer thereof.

In the present invention, the term "improve skin wrinkles" means maintaining or enhancing the ability to be associated with wrinkles and elasticity of the skin. Collagen (collagen) and elastic fiber elastin (collagen) in the dermal layer of the skin is the main protein that plays a role in the skin elasticity, collagen biosynthesis is affected by the internal and external skin. Specifically, the skin cells are reduced in cell activity due to natural aging, the collagen fibers are reduced, or the active oxygen produced by excessive irradiation of ultraviolet rays or stress as an external factor, the thiol group of the protein (thiol: -SH) In response to inhibiting the enzyme activity or by increasing the expression of degradation enzymes, such as collagen, elastin, increase the wrinkles of the skin and decrease the elasticity, the skin aging progresses.

In the present invention, the terms "cosmetic acceptable salt", "food acceptable salt", "pharmaceutically acceptable salt" or "salt thereof" may be an acid addition salt formed by free acid. have. Acid addition salts can be prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. In addition, equimolar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

As the free acid, an inorganic acid or an organic acid may be used. Non-limiting examples of the inorganic acid may be hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like, which may be used alone or in combination of two or more. Non-limiting examples of the organic acid are methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid (propionic acid). acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid (glucuronic acid), aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid and the like can be used. These may be used alone or in combination of two or more thereof.

In addition, the diosmin may use a base to make a cosmetically or food acceptable metal salt. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compounds salt, and then evaporating and drying the filtrate. As the metal salt, it is particularly preferable to prepare sodium, potassium or calcium salts, but is not limited thereto. Corresponding silver salts can also be obtained by reacting an alkali or alkaline earth metal salt with a suitable silver salt (eg silver nitrate).

The salts of diosmin may include all salts of acidic or basic groups which may be present in the compound of diosmin unless otherwise indicated. For example, the salt of the diosmin may include sodium, calcium and potassium salts of the hydroxy group, and other cosmetically acceptable salts of the amino group include hardbromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, Dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, and the like. It can be prepared through the method.

The effective amount of diosmin or its cosmetically acceptable salt in the cosmetic composition of the present invention is not particularly limited, and may be included in 0.0001 to 20% by weight based on the total weight of the composition. Less than 0.0001% by weight of diosmin or its salt in the cosmetics may have a small amount of anti-wrinkle effect, and more than 20% by weight of diosmin or its salt may exhibit known toxicity.

In one embodiment of the present invention, the procollagen type I C-peptide (PIP) secretion was increased when compared to the control cells irradiated with ultraviolet rays only when treated with diosmin in the human dermal fibroblasts irradiated with ultraviolet rays (Fig. 1).

In another embodiment of the present invention by supplying the experimental diet of Table 1 containing diosmin to the hairless mice for 10 weeks with ultraviolet irradiation, skin moisture content, water evaporation, elasticity, erythema index, back skin tissue of the experimental animal Wrinkles, changes in skin thickness, changes in the amount and morphology of collagen fibers, collagen types 1α1, 1α2, 3α1 and MMP-1a, MMP-1b, MMP-3, MMP-9 gene expression were measured. As a result, it was confirmed that diosmin significantly increased the skin moisture content and elasticity, and significantly reduced the water vaporization and erythema index (FIG. 3). In addition, diosmin improves wrinkles of the back skin tissues of experimental animals (FIG. 4), and significantly reduces skin wrinkle area, depth and length, significantly inhibiting wrinkle formation (FIG. 5). The thickness of the epidermal layer thickened by this was also significantly reduced (FIG. 6), and the effect of densifying collagen fibers and maintaining the arrangement regularly (FIG. 7), significantly reducing the expression of collagen types 1α1, 1α2 and 3α1. Increase the synthesis of collagen protein and significantly inhibit the degradation of collagen fibers by effectively reducing the expression of MMP-1a, MMP-1b, MMP-3, and MMP-9 genes, effectively inhibiting wrinkle formation by UV irradiation It was confirmed (Fig. 8).

Therefore, the composition comprising the diosmin of the present invention or a salt thereof is a functional cosmetic that provides an effect of improving skin wrinkles, improving skin moisturizing, removing skin dead skin, enhancing skin elasticity, inhibiting erythema, improving skin wrinkles and / or skin photoaging, It can be usefully used as a material for health functional food, medicine or quasi-drugs.

As used herein, the term "cosmetic composition" is a composition comprising the compound, the formulation may be in any form. For example, the cosmetic preparation prepared using the composition may be a nourishing cream, eye cream, massage cream, cream such as cleansing cream, pack, lotion such as nutrient lotion, essence, softening lotion, and lotion such as nourishing lotion. , Powders, foundations, makeup bases, and the like, and may be prepared and commercialized in any of these formulations to achieve the object of the present invention, and are not limited to the above examples. In addition, the cosmetic composition according to the present invention can be formulated by a conventional cosmetic preparation method.

Specifically, the cosmetics of the present invention include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, pack, mask pack, mask sheet It may be one having a formulation selected from the group consisting of, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder and eye shadow.

The cosmetic composition of the present invention may include other additives such as excipients, carriers, etc. in addition to diosmin or salts thereof, and it is possible to apply and formulate as needed the usual ingredients to be used in general skin cosmetics.

Specifically, the cosmetic composition of the present invention may further include a transdermal penetration enhancer. As used herein, the term transdermal penetration enhancer is a composition that allows a desired component to penetrate into the blood vessel cells of the skin at a high absorption rate. Preferably other phospholipid components, liposome components and the like used in lecithin cosmetics are included, but are not limited to these.

In addition, as the oil which can be mainly used as an oil phase component, one or more selected from vegetable oil, mineral oil, silicone oil and synthetic oil can be used. More specifically, mineral oil, cyclomethicone, squalane, octyldodecyl myristate, olive oil, Vitis binifera seed oil, macadamia nut oil, glyceryl octanoate, castor oil, ethylhexyl isononanoate, dimethicone Chicon, cyclopentasiloxane, sunflower seed oil and the like can be used.

In addition, 0.1 to 5% by weight of a surfactant, a higher alcohol, and the like may be added to reinforce the emulsifying ability. Such surfactants may be used conventional surfactants such as nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, phospholipids, and the like, specifically, sorbitan sesquinolate, polysorbate 60 , Glyceryl stearate, lipophilic glyceryl stearate, sorbitan oleate, sorbitan stearate, die-cetyl phosphate, sorbitan stearate / sucrosecoate, glyceryl stearate / polyethylene glycol-100 Stearate, ceteareth-6 oleate, arachidyl alcohol / behenyl alcohol / arachidyl glucoside, polypropylene glycol-26-butes-26 / polyethylene glycol-40 hydrogenated castor oil, etc. have. As the higher alcohol, alcohols having 12 to 20 carbon atoms, such as cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol, etc. may be used alone or in combination of one or more thereof.

The aqueous phase component may further add 0.001 to 5% by weight of one or more thickeners such as carbomer, xanthan gum, bentonite, magnesium aluminum silicate, cellulose gum, dextrin palmitate and the like to adjust the viscosity or hardness of the aqueous phase.

In addition, the cosmetic composition of the present invention, if necessary, active ingredients such as higher fatty acids, vitamins, sunscreens, antioxidants (butylhydroxyanisole, propyl gallic acid, elixolic acid, tocopheryl acetate, butylated hydroxy) Toluene), preservatives (methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinylurea, chlorphenesin, etc.), colorants, pH adjusters (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, Sodium malic acid, fmaric acid, sodium pramate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, etc., moisturizers (glycerine, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin , Betaine, glycerin-26, methylgluse-20 and the like), lubricants and the like can be further added.

In addition, the cosmetic composition of the present invention further comprises a substance capable of auxiliaryly providing essential nutrients to the skin, and may preferably contain auxiliary agents including, but not limited to, natural flavors, cosmetic flavors, or herbal medicines. have.

In addition, the cosmetic composition of the present invention may further comprise a skin wrinkle improvement component or skin elasticity enhancing component. The specific skin wrinkle improving component or skin elasticity enhancing component may be any one or more selected from the group consisting of vitamin C, retinoic acid, TGF, protein from animal placenta, betulinic acid and chlorella extract, most preferably Vitamin C.

In one embodiment of the present invention, when treated with vitamin C in the human skin fibroblasts irradiated with ultraviolet rays, the amount of collagen is measured to a higher value than the case of treating diosmin or vitamin C alone, synergistic effect It was confirmed that it is shown (Fig. 1).

The present invention also provides a health function having a skin moisturizing improvement, skin exfoliation, skin elasticity enhancement, erythema suppression, skin wrinkle improvement and / or skin photoaging improvement effect, which contain diosmin or a food acceptable salt thereof as an active ingredient. Provide a food composition.

The description of "diosmin" and "food acceptable salt thereof" is as described above.

In the present invention, the term "health functional food" refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials or ingredients having useful functions for the human body. Here, 'functional' means to obtain a useful effect for health purposes such as nutrient control or physiological action on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding raw materials and ingredients commonly added in the art. In addition, the formulation of the health functional food can also be prepared without limitation as long as the formulation is recognized as a health functional food. The health functional food composition of the present invention has the advantage that there is no side effect that can occur when taking long-term use of the drug, unlike foods as a raw material, and excellent portability, improve skin moisturizing, skin exfoliation, skin elasticity Ingestion may be taken as an adjuvant to promote erythema suppression, skin wrinkle improvement and / or skin photoaging improvement.

The dietary supplement composition of the present invention may be prepared in any one formulation selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and rings.

Health functional food composition according to the present invention may be formulated in the form of powder, liquid, tablets, soft capsules, granules, tea bags, instant tea or drink by including diosmin as an active ingredient. The content of diosmin as an active ingredient can be suitably determined according to the purpose of use (prevention or improvement). In general, the amount of diosmin included in the nutraceutical composition may be added to 0.1 to 90% by weight of the total food weight. However, in the case of prolonged intake for health and hygiene purposes or health control purposes, the amount may be below the above range. In addition, the nutraceutical composition according to the present invention, in addition to diosmin, other ingredients that can give a synergistic effect to the main effect within the range that does not impair the main effect of the present invention, for example, vitamin C and It may contain such compounds for improving wrinkles or natural products, such as green tea extract, mulberry extract, licorice extract, lettuce extract, betel nut extract, golden extract, wild ginseng extract.

The dietary supplement composition formulated in the form as described above may be added to the food as it is or used with other foods or food ingredients, and may be appropriately used according to conventional methods. Examples of foods include drinks, meats, sausages, breads, biscuits, rice cakes, chocolate, candy, snacks, confectionery, pizza, ramen, dairy products including other noodles, gums, ice creams, various soups, beverages, alcoholic beverages and vitamin complexes. , Dairy products and dairy products, and all functional foods in the conventional sense.

When the health functional food composition of the present invention is a drink, it contains diosmin as an essential ingredient in the ratio indicated, and there are no particular restrictions on other ingredients used for the purpose of preparing other drinks, and various flavoring agents such as ordinary drinks. Or natural carbohydrate or the like as an additional component. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a flavoring agent other than the above-mentioned, a natural flavoring agent, a synthetic flavoring agent, etc. can be used. The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition, the nutraceutical composition of the present invention is a variety of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain a pulp for producing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the diosmin of the present invention.

The present invention also provides a pharmaceutical or quasi-drug composition having a skin moisturizing improvement, skin exfoliation, skin elasticity improvement, erythema suppression, wrinkle improvement and / or skin photoaging improvement effect containing diosmin or a salt thereof as an active ingredient. .

The description of "diosmin" and "salts thereof" is as described above.

When using the diosmin of the present invention or a pharmaceutically acceptable salt thereof as a medicine, it may further contain one or more active ingredients exhibiting the same or similar functions. For example, it may include known skin wrinkle improving ingredients or elasticity enhancing ingredients. By including additional skin wrinkle improving component and elasticity enhancing component, the wrinkle improvement and elasticity enhancing effect of the composition of the present invention can be further enhanced. When the ingredient is added, skin safety, ease of formulation, and stability of the active ingredients may be considered. In one embodiment of the present invention, the pharmaceutical composition is a wrinkle improving component known in the art, any one or more selected from the group consisting of vitamin C, retinoic acid, TGF, protein from animal placenta, betulinic acid and chlorella extract It may further comprise a skin wrinkle improving ingredient. The additional skin wrinkle improving component may be included in 0.0001 to 10% by weight relative to the total weight of the composition, the content range is based on requirements such as collagen synthesis promoting activity, skin safety, ease of formulation of the compound of Formula 1 Can be adjusted accordingly.

In addition, the pharmaceutical composition for improving skin wrinkles and elasticity of the present invention may further comprise a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain various components such as buffers, sterile water for injection, general saline or phosphate buffered saline, sucrose, histidine, salts, polysorbates and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and may be administered in the form of a general pharmaceutical preparation, for example, in various formulations, orally and parenterally, in the case of clinical administration. , Diluents or excipients such as extenders, binders, wetting agents, disintegrants, surfactants and the like.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the pharmaceutical composition of the present invention, for example, starch, calcium carbonate, It may be prepared by mixing sucrose or lactose, gelatin and the like.

In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The pharmaceutical composition of the present invention may provide a desirable skin moisturizing improvement, skin exfoliation, skin elasticity improvement, erythema suppression, wrinkle improvement or skin photoaging improvement when including an effective amount of diosmin or a pharmaceutically acceptable salt thereof. have. In the present invention, the term 'effective amount' means an amount of a compound capable of inhibiting or inhibiting generation of wrinkles on the skin or alleviating wrinkles already produced. The effective amount of diosmin or a pharmaceutically acceptable salt thereof contained in the composition of the present invention will vary depending on the form in which the composition is commercialized, the method by which the compound is applied to the skin and the time it stays on the skin. For example, when the composition is commercialized as a medicament for dermatological treatment due to wrinkle formation, elasticity reduction, and blemishes of the skin, diosmin or a pharmaceutical composition thereof is present at a higher concentration than when it is commercialized as a cosmetic product that is applied to the skin on a daily basis. It may include an acceptable salt.

The term "quasi drug" used in the present invention refers to articles that have a lesser action than drugs among those used for the purpose of diagnosing, treating, ameliorating, alleviating, treating or preventing diseases of humans or animals. According to the quasi-drug product, the drug is used for the purpose of medicine, and it includes a product used for the treatment or prevention of diseases of humans and animals, and a product having a slight or no direct action on the human body.

The quasi-drug composition of the present invention is used for the purpose of improving skin moisturizing, removing skin dead skin, enhancing skin elasticity, suppressing erythema, improving skin wrinkles and / or improving skin photoaging, and is not particularly limited in the formulation thereof. It may be a cosmetic composition having a formulation of a flexible cosmetics, nourishing cosmetics, massage creams, nutrition creams, packs, mask packs, mask sheets, gels or skin adhesive cosmetics, and also lotions, ointments, gels, creams, patches or sprays. It may be a transdermal dosage form such as

In addition, in each formulation, the quasi-drug composition may be optionally selected and blended with other ingredients according to the formulation or purpose of use of other quasi-drugs. The mixed amount of the active ingredient can be suitably determined depending on the purpose of use (inhibition or relaxation). For example, it may include conventional adjuvants such as thickeners, stabilizers, solubilizers, vitamins, pigments and flavorings, carriers and the like.

The content of the diosmin or salt thereof of the present invention is preferably 0.0001 to 20% by weight, respectively, based on the total weight of the quasi-drug composition. If the content exceeds 20% by weight, the color and stability of the composition are inferior. If the content is less than 0.0001%, the effect is minimal.

In addition, in the quasi-drug composition of each formulation, other components than the essential components described above can be appropriately selected and blended by those skilled in the art without difficulty depending on the formulation or purpose of use.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Example 1

Human skin fibroblasts  Used Diosmin  Wrinkle improvement effect

1-1. Experiment method

1) Cell Culture

Human dermal fibroblasts (normal, human, neonatal, ATCC No. PCS-201-010) were purchased from ATCC (Manassas, VA, USA). The purchased cells were cultured in a 37, 5% CO 2 incubator using a fibroblast growth medium (Promo Cell, Heidelberg) was used for the experiment.

2) Procollagen Type I C-peptide (PIP) concentration measurement

A human fibroblasts In order to examine the collagen biosynthesis performance 12-well-was added to the plate (well-plate) 1.0 × 10 6 cells / by well Pipette DIOS min and vitamin C at a concentration of 100 μM respectively for 24 hours, CO ₂ Cultured in the incubator. After removing the medium of each well, washed once with PBS, and again put 1 ml of PBS was irradiated with ultraviolet B (UVB) at 20 mJ / cm ² conditions. PBS of each well was replaced with medium again and cultured for 24 hours, and then the amount of procollagen secreted into the medium was measured using a procollagen type C-peptide EIA kit (Takara Bio, Japan). The standard solution included in the collagen measurement kit was diluted by concentration, and the absorbance was measured at 450 nm to prepare a standard concentration curve and calculate the amount of collagen produced.

1-2. Experiment result

1) Changes in Procollagen Secretion

Collagen, the main protein constituting the skin, is synthesized in the form of procollagen from fibroblasts present in the dermis and then secreted into the extracellular matrix. Procollagen secreted into the extracellular matrix is decomposed by the procollagen peptidase on the cell surface and formed into active collagen. Therefore, the activated collagen content can be determined by measuring the C-peptide content. Each value in FIG. 1 represents the means ± SEM of three measurements from three independent wells. Mean values in other alphabets indicate statistical significance ( p <0.05).

Human skin fibroblasts were treated with UV radiation and treated with drugs to measure the amount of procollagen and procollagen type I C-peptide (PIP) secreted into the extracellular matrix. Procollagen secretion was significantly reduced compared to -UVB), and the amount of collagen was increased 21% in UV-irradiated cells treated with diosmin (DM) compared to UV-irradiated control cells (+ UVB). On the other hand, cells treated with diosmin with vitamin C (VitC + DM) showed a 45% significant increase in the amount of collagen compared to UV control cells (+ UVB). This is higher than the amount of collagen observed in cells treated with either vitamin C (+ 27%) or diosmin (+ 21%) alone (FIG. 1). Substituting these results into Colby's formula, it can be seen that synergistic effects are obtained when diosmin and vitamin C are treated together. Therefore, diosmin increases the amount of collagen activated in human skin fibroblasts, and this collagen increase effect is more effective when used with vitamin C.

Example 2

Using mouse Diosmin  Skin wrinkle improvement, moisturizing and elasticity efficacy evaluation

2-1. Experiment method

1) Preparation of experimental diet, breeding of experimental animals and UV irradiation

The five-week-old female albino hairless mice (Skh-1) used in this example were purchased from Orient Bio (Gyeonggi-do, Korea) and subjected to a one-week adaptation period with solid feed. Experimental animals were divided into 4 groups and 5 animals were assigned to each group and used for the experiment. All experimental groups were divided into the normal control group (-UVB), the ultraviolet irradiation group (+ UVB), and the ingestion of diosmin (CV) or vitamin C (VitC) with ultraviolet irradiation. During the breeding period, feed and water were freely ingested, and the temperature was maintained at 22 ± 1 and the humidity at 60 ± 5%, and the photoperiod and dark cycle were adjusted to 12 hours daily.

The -UVB and + UVB groups consumed a refined diet prepared according to the AIN-93 rodent diet (Reeves, PG et al., J Nutr, 123: 1939-1951, 1993). A diet prepared by adding 0.2% diosmin (Sigma-Aldrich) and the VitC group to the AIN-93 tablet diet by adding 0.2% vitamin C (Sigma-Aldrich) for 10 weeks. The composition of the detailed experimental diet is shown in Table 1 below. The diet was fed with water between 10 am and 11 am daily, and dietary intake was measured daily.

Experimental composition ingredient AIN- 93G diet (g / kg diet) Diosmin Supplementary diet (DM) (g / kg diet) Vitamin c Supplementary diet ( Vitc ) (g / kg diet) Casein 200 200 200 Maltodextrin 132 132 132 Corn starch 397.486 395.486 395.486 Sucrose 100 100 100 cellulose 50 50 50 Soybean oil 70 70 70 Vitamin complex 10 10 10 Mineral complex 35 35 35 Choline Bitartrate 2.5 2.5 2.5 L-cystine 3 3 3 Tert-Butyhydroquinone 0.014 0.014 0.014 Diosmin - 2 - Vitamin c - - 2 Total (g) 1,000 1,000 1,000

During the breeding period, ultraviolet B (UVB) was irradiated to the back of the hairless mice three times a week. The UV dose was 73 mJ / cm ² for the first week, 146 mJ / cm ^{2 for the} second week, and from 3 to 10 weeks. Irradiation at 219 mJ / cm ² . During breeding, body weight and skin thickness were measured every week, and photographs of the back skin were taken. Skin thickness was measured in the hip region of hairless mice using a digital micro caliper (Marathon Watch Company Ltd, Ontario, Canada). The caliper used in the measurement was able to measure up to 0.01 mm, and it has a control function to apply a constant force to the thickness, so that the thickness of the skin was measured under the same force.

On the last day of the experiment, the animals were fasted for 6 hours and blood and back skin tissues were collected under anesthesia. Blood samples were collected from the abdominal inferior vena cava with a syringe, centrifuged at 2,000 × g for 15 minutes, and plasma was separated and stored frozen at −70 until analysis. Some of the back skin tissues were immediately stored in -70 freezers and used for molecular biological tests, and some were fixed in 10% formalin solution and used for immunohistochemical staining.

2) skin moisturizing, elasticity and Erythema index  Measure

Skin moisture content, water evaporation, elasticity, and erythema index were measured once using the Corneometer®, Tewameter®, Cutometer®, and Mexameter® (CK Electronics GmbH), respectively. When measuring, the numerical value that appears by lightly pressing a certain part of the experimental animal and the like was recorded.

3) Dorsal skin tissue  Wrinkle measurement

The skin of hairless mice subjected to UV irradiation for 10 weeks was made of silicone rubber to measure the extent of wrinkle formation. Attach a disk with a circular hole with a diameter of 1 cm to the back of the hairless mouse, mix the reagent for making a replica, thinly spread it on the back of the hairless mouse, dry it completely, and carefully remove the disk. Was produced. Fabrication temperature of the replica plate was carried out in a constant temperature and humidity condition of 20 ~ 23, humidity of 45 ~ 50%, and a silicon rubber impression material (Epigem, Seoul, Korea) for producing a replica plate. Analysis of the simulated platen was performed using a computer image analyzer (Visioline VL650, CK electronic GmbH, Germany) for total wrinkle area, maximum wrinkle depth, mean depth and mean wrinkles. Four wrinkle indicator items such as mean length were analyzed.

4) Immunohistochemical staining of skin tissue

The back skin tissue of hairless mice was extracted, fixed in 10% formalin, and then stained with Hematoxylin and eosin (H & E) and Masson's trichrome (M & T) by a Korean CFC (Gyeonggi-do, Korea), followed by an optical microscope (IX71, Olympus, JPN). ), And photographed using a digital camera (DP71, Olympus, JPN).

5) RT- PCR  analysis

Tissue was ground by adding 1 ml of Trizol solution per 0.1 g of back skin tissue, and centrifuged at 4, 12,000 xg for 10 minutes. The supernatant was transferred to a new tube and 200 μl of chloroform was added and vortexed. After repeating this process twice, the supernatant was transferred to a new tube and isoprophanol and supernatant were added at a 1: 1 ratio. After shaking vigorously 10 times and left at room temperature for 15 minutes, centrifugation was performed at 12,000 xg and 4 minutes for 10 minutes, the supernatant was removed, and 1 ml of 70% ethanol was added to the remaining precipitate, followed by 5 minutes at 7,500 xg and 4 minutes. Centrifugation was performed. After removing ethanol, the tube containing the RNA precipitate was dried at room temperature for 15 minutes, and the RNA pellet was dissolved using nuclease free water. Using a UV / VIS spectrophotometer (Beckman coulter, DU730) was measured the concentration of RNA samples extracted at 260 nm and 280 nm wavelength, and the integrity of the RNA samples were confirmed by agarose gel electrophoresis.

CDNA was synthesized by reverse transcription using oligo dT primer and superscript reverse transcriptase (GIBCO BRL, Gaithersburg, MD, USA) on RNA samples extracted from the back skin tissue. PCR was performed using 5D and 3 'flanking sequences of the gene cDNA to be amplified as a template and cDNA obtained through reverse transcription, and the primer sequences used are shown in Table 2. 1 μl of the amplified PCR product was electrophoresed on 1% agarose gel to confirm DNA band.

Primer sequence used for RT-PCR gene primer Sequence (5 '→ 3') Annealing Temperature (℃) PCR product (bp) Collagen type alpha1 (Col1α1) F ggcaacagtcgcttcaccta (SEQ ID NO: 1) 55 164 R agtccgaattcctggtctgg (SEQ ID NO: 2) Collagen type alpha2 (Col12) F cggttctgttggtcctgttg (SEQ ID NO: 3) 55 103 R acccctgtgccctttatcac (SEQ ID NO: 4) Collagen type alpha1 (Col3α1) F taaccaaggctgcaagatgg (SEQ ID NO: 5) 55 104 R accagtgcttacgtgggaca (SEQ ID NO: 6) Matrixmetallopeptidase 1a (MMP-1a) F ccctgtgtttcacaacggag (SEQ ID NO: 7) 55 133 R cctcagcttttcagccatca (SEQ ID NO: 8) Matrixmetallopeptidase 1b (MMP-1b) F tttgctcatgcttttctgcc (SEQ ID NO: 9) 55 146 R gaatgggagagtccaaggga (SEQ ID NO: 10) Matrixmetallopeptidase 3 (MMP-3) F tgctggtatggagcttctgc (SEQ ID NO: 11) 55 142 R catctccaacccgaggaact (SEQ ID NO: 12) Matrixmetallopeptidase 9 (MMP-9) F gtggaccatgaggtgaacca (SEQ ID NO: 13) 55 102 R actgcacggttgaagcaaag (SEQ ID NO: 14) Glyceraldehyde3-phosphatedehydrogenase (GAPDH) F ggagattgttgccatcaacg (SEQ ID NO: 15) 55 122 R tgacaagcttcccattctcg (SEQ ID NO: 16)

2-2. Experiment result

1) body weight of hairless mice and Dietary Intake

Ultraviolet irradiation, diosmin and vitamin C intake did not significantly affect the body weight and dietary intake of hairless mouse mice (FIG. 2). The values in FIG. 2 are mean ± SEM of 5 mice.

2) moisture content, elasticity and Erythema index  change

The + UVB control group irradiated with UV light for 10 weeks significantly reduced the moisture content and elasticity of skin tissues compared to the normal group (-UVB) without UV irradiation, while the water evaporation and erythema index increased significantly (Fig. 3). In the DM group fed with diosmin, moisture content and elasticity were significantly increased by 69% and 57%, respectively, compared to the + UVB control group. % And 44% were found to decrease significantly. The water content and elasticity enhancing effect of this diosmin was similar to the effect by vitamin C (Fig. 3). The values in FIG. 3 are mean ± SEM of 5 mice, and the alphabet above the bar represents a significant difference by one-way ANOVA followed by Duncan's multiple range test (p <0.05).

3) Changes in Wrinkle Formation in Hairless Mice with UV Irradiation

To evaluate the effect of diosmin intake on the formation of skin wrinkles, photographs of the back skin of experimental animals were taken. The + UVB control group, which had been irradiated with UVB for 10 weeks, was able to visually observe the formation of fine wrinkles with a number of coarse and deeply wrinkled wrinkles compared to the normal group (-UVB), which was not irradiated with UVB. Compared with the + UVB control group, the thickness and depth of the wrinkles were significantly reduced, which was similar to the skin condition observed in the -UVB group not irradiated with ultraviolet rays (FIG. 4).

10 weeks after the UV-irradiated hairless mice were made with silicone rubber to measure the extent of wrinkle formation, the + UVB control group was thicker and deeper than the normal UV-irradiated group (-UVB). It was observed that fine wrinkles were formed together with the wrinkles, and the diosmin intake group showed a significant improvement in the thickness and depth of the wrinkles, despite the fact that the UV rays of the same intensity were irradiated with the deep wrinkles almost disappearing compared to the + UVB control group. (FIG. 5A). Even in the result of quantifying wrinkle formation in computer simulation using the computer image analyzer, DM group had 70% total wrinkle area, maximum wrinkle depth 59%, average wrinkle depth 29%, and average wrinkle compared to + UVB group. The length was significantly reduced by 58%, and the antiwrinkle effect of diosmin was similar to the antiwrinkle effect observed in vitamin C (FIG. 5B). The values in FIG. 5B are mean ± SEM of 5 mice, and the alphabet above the bar represents a significant difference by one-way ANOVA followed by Duncan's multiple range test (p <0.05). Therefore, it can be seen that ingestion of diosmin has an effect of significantly inhibiting wrinkle formation by ultraviolet irradiation.

4) Changes in skin thickness of UV-radiated hairless mice

As photoaging progresses due to ultraviolet rays, the formation of the stratum corneum increases to protect the dermal layer of the skin, resulting in thickening of the skin. Therefore, the thickening of the skin epidermal layer by UV irradiation means that the skin damage caused by photoaging is large. J Molecular mechanism of skin ageing Mech Ageing Dev 123: 801-810, 2002)

As a result of measuring the thickness of the back skin by using digital micro caliper on the last day of the experiment, the + UVB control group significantly increased the skin thickness compared to the normal group (-UVB) without UV irradiation. It was confirmed that the skin thickness was significantly reduced by 23% compared to the + UVB control (Fig. 6A). The values in FIG. 6A are mean ± SEM of 5 mice, with the alphabet above the bar representing a significant difference by one-way ANOVA followed by Duncan's multiple range test (p <0.05).

The skin epidermal layer thickness of hairless mice by H & E staining of skin tissue was also observed that the + UVB control group thickened the skin epidermis layer compared to the normal group (-UVB) without UV irradiation. It was confirmed that the thickness of the epidermal layer thickened by was significantly reduced (FIG. 6B).

5) Changes in the amount and morphology of collagen fibers in the dermis of UV-radiated hairless mice

In the case of skin that has undergone photoaging by UV irradiation, the collagen fiber network at the epidermal-dermal junction is damaged, and thus the amount and form of the collagen fiber is used as an indicator of skin aging (Varani et al. "Inhibition of type I procollagen synthesis by damaged collagen in photoaged skin and by collagenase-degraded collagen in vitro. "The American journal of pathology 158: 931-942, 2001). The skin tissue of UV-radiated hairless mice was stained with collagen fibers by Masson's trichrome staining. As a result, collagen fibers strongly stained with aniline blue were observed in the dermal layer under the epidermis of -UVB group. After 10 weeks of irradiation, collagen fibers of + UVB group were weakly stained by aniline blue. In the dermal layer of UV-irradiated diosmin for 10 weeks, the density of collagen fibers was more dense and orderly compared to the control group, and the staining by aniline blue was also stronger. (FIG. 7).

6) Gene Expression Changes in Skin Tissue of Ultraviolet Irradiated Hairless Mice

Collagen types 1 and 3 are proteins that constitute the cytoplasmic components of the dermal layer, and in particular, type 1 collagen is present in the largest amount of extracellular matrix proteins present in skin connective tissue. On the other hand, there are 23 types of MMPs that catalyze collagen degradation, and MMP types known to increase by UV rays are known as Nos. 1, 3, and 9. These three types of MMPs are enzymes that degrade collagen types 1 and 3. to be. MMP-1 is known to cut the middle of collagen fibers, while MMP-3 and MMP-9 are known to play a role in subdividing and cutting the cut collagen fibers (Quan T. et al. Elevated matrix metalloproteinases and collagen fragmentation). in photodamaged human skin: impact of altered extracellular matrix microenvironment on dermal fibroblast function.J Investigative Dermatology, 133: 1362, 2013; Kondo S. The roles of cytokines in photoaging.J Dermatological Science 23: S30-S36, 2000).

As a result of evaluating gene expression changes by RT-PCR analysis on skin tissue, collagen type 1α1 and α2 and collagen type 3α1 of + UVB control group were compared with normal group (-UVB) without UV irradiation. The expression of MMP-1a and -1b, MMP-3, and MMP-9 genes were significantly increased. On the other hand, the expression of collagen type 1α1 and α2 and collagen type 3α1 was significantly increased in the diosmin intake group compared to the + UVB control group, and the expression of MMP-1a and -1b, MMP-3, and MMP-9 genes was significantly increased. Decrease (Fig. 8). Therefore, it is thought that diosmin suppressed wrinkle formation by ultraviolet irradiation by increasing collagen protein synthesis of skin tissue and inhibiting collagen fiber degradation.

The above results suggest that diosmin may have an effect to prevent or alleviate skin wrinkles caused by ultraviolet rays, which may be usefully used as functional cosmetics, health functional foods, medicines or quasi-drugs in the future.

Example 3

Skin wrinkle improvement, skin irritation and dead skin removal sensory test

3-1. Diosmin  Containing nutrition cream ( Experimental Example 1) and Diosmin  Nourishing cream Comparative example 1)  Produce

number ingredient Experimental Example 1 (% by weight) Comparative Example 1 (% by weight) One Cetearyl Alcohol 1.5 1.5 2 Glyceryl Stearate One One 3 Polysorbate 60 1.2 1.2 4 Sorbitan sesquioleate 0.3 0.3 5 Cetyloctanoate 6 6 6 Squalane 8 8 7 Apricot Kernel Oil 4 4 8 Dimethicone One One 9 Butylene Glycol 5 5 10 glycerin 4 4 11 Magnesium Aluminum Silicate 0.2 0.2 12 Xanthan Gum 0.05 0.05 13 antiseptic a very small amount a very small amount 14 Purified water Remaining amount Remaining amount 15 Diosmin One -

Among the components identified by each component number in Table 3 above, components 1 to 8 are first dissolved by heating at a temperature of 70, and then components 9 to 13 are dissolved and dispersed in component 14 and emulsified to 70. Thereafter, the emulsified product was cooled to a temperature of 56, and then, component 15 dissolved in the fractionated component 9 was added, stirred, and cooled to room temperature to prepare.

Comparative Example 1 with respect to Experimental Example 1, except for the component 15, diosmin, except for the component composition and the manufacturing method was set to proceed in the same manner.

3-2. Wrinkle improvement effect and skin irritation sensory test

In order to evaluate the skin wrinkle improvement effect and skin irritation of the cosmetic composition according to the present invention, the sensory test was carried out using the nutrition cream prepared in Experimental Example 1 and Comparative Example 1 of Table 3.

Specifically, in order to measure the antiwrinkle effect of the skin when the nutrition cream of Experimental Example 1 and Comparative Example 1 formulations described in Table 3 were applied to the skin, Experimental Example 1 nutrition cream (test group) of 3, Comparative Example 1 nutrition cream (control group) of Table 3 to the right side of the face was to be used continuously for 12 weeks once a day.

In the sensory test, the anti-wrinkle effect items of the skin were evaluated to evaluate the anti-wrinkle effect of the nutritional cream of Experimental Example 1 based on the nutritional cream of the comparative example. This phenomenon was evaluated. Evaluation was performed based on the five-point standard of very good (5 points), excellent (4 points), moderate (3 points), bad (2 points), very bad (1 point), the results are shown in Table 4 below. Indicated. Skin stimulation in Table 4 indicates that the higher the score, the less skin stimulation.

number Skin irritation wrinkle improvement Experimental Example 1 Comparative Example 1 Experimental Example 1 Comparative Example 1 One 5 4 3 3 2 5 5 4 4 3 4 4 5 4 4 4 4 5 5 5 5 5 4 3 6 4 5 4 4 7 4 4 4 4 8 4 5 4 5 9 3 4 4 4 10 4 5 5 4 11 5 4 4 4 12 5 5 5 4 13 4 4 5 4 14 5 5 4 4 15 4 3 3 3 16 5 5 4 3 17 5 4 4 4 18 5 5 4 4 19 4 4 5 5 20 4 4 5 5 Average 4.4 4.4 4.25 4.0

As shown in Table 4, the skin irritation evaluation score for the nutritional cream formulation of Experimental Example 1 containing diosmin was evaluated very well at 4.4, and the skin was the same as the nutritional cream formulation of Comparative Example 1 which received the same score. The low degree of irritation was confirmed to be excellent skin safety.

In addition, the relative skin wrinkle improvement effect of the nutritional cream formulation of Experimental Example 1 containing diosmin compared to the nutritional cream formulation of Comparative Example 1 was 4.25 points, it can be seen that the degree of wrinkle improvement is also very excellent.

Therefore, the cosmetic composition containing the diosmin of the present invention or its cosmetically acceptable salt as an active ingredient has no skin side effects, improves wrinkles, increases the synthesis of collagen protein, and inhibits the degradation of collagen fibers. Excellent for wrinkle improvement

3-3. skin Moisturizing  Increase measurement test

In order to measure the moisture retention ability of the skin when the nutrition cream of the Experimental Example 1 and Comparative Example 1 formulations described in Table 3 were applied to the skin, the left side of the face was shown in Table 3 in 20 women in their 20s to 50s. Experimental Example 1 The nourishing cream (test group) was applied to the right side of the face by gently massaging the Comparative Example 1 nourishing cream (control group) of Table 3 in a sufficient amount. Massage was evenly carried out over about 1 minute.

The skin moisture content was measured using a Koniometer in a constant temperature and humidity room at room temperature 25 and relative humidity 45% before application, and the Experimental Example 1 and Comparative Example 1 formulations of Table 3 were placed on the left and right sides of the face, respectively. The skin was rested for 10 minutes after application and the skin moisture content was measured 12 hours after application and 24 hours after application. The measuring device measured the moisture content of the skin using a moisture content meter (coneometer; CM820, Courage Khazaka electronic GmbH, Germany) that measures the moisturizing power by measuring the electric capacity of the skin according to the moisture content of the skin. The conniometer measures the moisture level by measuring the amount of moisture by measuring the amount of water ions using a sensor and measuring the amount of moisture present in the skin epidermis, and the measuring method is as follows.

1) The Koniometer probe was placed on the skin area to be measured.

2) When the probe is pressed on the skin, the electric conductivity of the skin through the sensor is numerically displayed on the screen.

3) The measurement was repeated with different measurement sites.

4) After measuring once, the sensor was wiped with a tissue such as Kimwipe and then measured again.

The measured values are shown in Table 5 below as average values.

Test substance Skin moisture content (%) Before application 12 hours later 24 hours later Experimental Example 1 30.3 44.1 38.7 Comparative Example 1 30.2 38.0 30.8

As shown in the above results, when the nutrition cream of the comparative example was applied, after 12 hours of application, the skin moisture content increased to some extent, but after 24 hours, it can be seen that it returned to a level similar to the state before application. On the other hand, when the nutrition cream of Experimental Example 1 containing the diosmin was applied to the skin, it was confirmed that the moisturizing power was maintained for more than 24 hours and the skin moisture increase effect was also higher than that of Comparative Example 1.

3-4. Exfoliation effect test

In order to measure the skin exfoliation effect when the nourishing creams of the Experimental Example 1 and Comparative Example 1 formulations described in Table 3 were applied to the skin, respectively, 20 women in their 20's to 50's ages had the left side of Table 3 Experimental Example 1 The nourishing cream (test group) was applied to the right side of the face by gently massaging the Comparative Example 1 nourishing cream (control group) of Table 3 in a sufficient amount. Massage was evenly carried out over about 1 minute.

The measurement of the angular mass of the skin was measured by using Charm view (Moritex, Japan) in a constant temperature and humidity room at room temperature 25 and relative humidity of 45% before application, and Experimental Example 1 and Comparative Example 1 formulation of Table 3 above. Were applied to the left and right sides of the face, respectively, and the skin angular mass after 24 hours was measured, and the amount of skin exfoliation was calculated. Each value is shown in Table 6 below as an average value.

Test substance Skin angular mass (%) Before application 24 hours later Decrease in dead skin cells Experimental Example 1 20.4 9.6 10.8 Comparative Example 1 20.2 15.9 4.3

As shown in the above results, it was confirmed that the nutrition cream of Experimental Example 1 containing diosmin is superior to the skin exfoliation effect than the nutritional cream of Comparative Example 1 containing no diosmin. As the moisturizing effect is applied to the skin, the angular mass decreases. The nourishing cream of Experimental Example 1 has a superior skin exfoliation effect than the nourishing cream of Comparative Example 1 due to its excellent water retention as demonstrated in Example 3-3. It is judged to indicate.

[ Production Example  1] Manufacture of Cosmetics

1-1. Preparation of Flexible Lotion

To the composition of Table 7 below, a flexible lotion containing diosmin was prepared.

Furtherance Production Example  1-1 ( weight% ) Diosmin 0.1 ethanol 10.0 Polylauric acid polyoxyethylene sorbitan 1.0 Paraoxybenzoic Acid Methyl 0.2 glycerin 5.0 1,3-butylene glycol 6.0 incense Quantity Pigment Quantity

1-2. Manufacture of nutritional lotion

To the composition of Table 8, a nourishing lotion containing diosmin was prepared.

Furtherance Production Example  1-2 ( weight% ) Diosmin 0.05 Waselin 2.0 Sesquioleate sorbitan 0.8 Polyoxyethylene oleylethyl 1.2 Paraoxybenzoic Acid Methyl Quantity Propylene glycol 5.0 ethanol 3.2 Carboxy Vinyl Polymer 18.0 Potassium hydroxide 0.1 Pigment Quantity incense Quantity

1-3. Manufacture of nutritional cream

With the composition of Table 9 below, a nutritive cream containing diosmin was prepared.

Furtherance Production Example  1-3 ( weight% ) Diosmin 0.2 Stearic acid 15.0 Cetanol 1.0 Potassium hydroxide 0.7 glycerin 5.0 Propylene glycol 3.0 antiseptic Quantity incense Quantity Purified water to 100

1-4. Manufacture of pack

With the composition shown in Table 10 below, a pack containing diosmin was prepared.

Furtherance Production Example  1-4 ( weight% ) Diosmin 0.05 glycerin 5.0 Propylene glycol 4.0 Polyvinyl alcohol 15.0 ethanol 8.0 Polyoxyethylene Oleethyl 1.0 Paraoxybenzoic Acid Methyl 0.2 Pigment Quantity incense Quantity

1-5. Preparation of Essence

With the composition shown in Table 11 below, an essence containing diosmin was prepared.

Furtherance Production Example  1-5 ( weight% ) Diosmin 0.2 Propylene glycol 10.0 glycerin 10.0 Sodium hyaluronate aqueous solution (1%) 5.0 ethanol 5.0 Polyoxyethylene Cured Castor Oil 1.0 Paraoxybenzoic Acid Methyl 0.1 incense Quantity Purified water to 100

[ Production Example  2] Preparation of health functional food

2-1. Preparation of health functional food

To the composition of Table 12, a dietary supplement containing diosmin was prepared.

Furtherance Production Example  2-1 Diosmin 1000 mg Vitamin mixtures Quantity Vitamin A Acetate 1.0 mg Vitamin E 0.13 mg Vitamin B1 0.15 mg Vitamin B2 0.5 mg Vitamin B6 0.2 μg Vitamin B12 10 mg Vitamin c 10 μg Biotin 1.7 mg Nicotinic acid amide 50 μg Folic acid 0.5 mg Calcium Pantothenate Quantity Mineral mixture Quantity Ferrous sulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium phosphate monobasic 15 mg Dibasic Potassium Phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg

Although the composition ratio of the vitamin and mineral mixture is a composition that is relatively suitable for the health functional food is mixed in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Next, the granules may be prepared and used for preparing the nutraceutical composition according to a conventional method.

2-2. Manufacture of healthy drinks

With the composition shown in Table 13 below, a health beverage containing diosmin was prepared.

Furtherance Production Example  2-2 (mg) Diosmin 1000 mg Citric acid 1000 mg oligosaccharide 100 g Plum concentrate 2 g Taurine 1 g Purified water Quantity

After mixing the above components in accordance with the conventional method for preparing healthy beverages, the solution prepared by stirring and heating at 85 for about 1 hour, filtered into 21 sterilized containers, sealed and sterilized and then refrigerated and stored Used to prepare beverage compositions. Although the composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

2-3. Manufacture of tablets

To the composition shown in Table 14, by tableting according to the conventional tablet manufacturing method to prepare a tablet containing diosmin.

Furtherance Production Example  2-3 mg Diosmin 100 mg Corn starch 100 mg Lactose 100 mg Magnesium Stearate 2 mg

2-4. Preparation of Capsules

To the composition shown in Table 15, according to the conventional capsule preparation method was filled in gelatin capsules to prepare a capsule containing diosmin.

Furtherance Production Example  2-4 (mg) Diosmin 100 mg Corn starch 100 mg Lactose 100 mg Magnesium Stearate 2 mg

2-5. Manufacture of rings

To the composition shown in Table 16, the ring was prepared so as to be 4 g per ring according to a conventional ring production method.

Furtherance Production Example  2-5 (mg) Diosmin 1 g Lactose 1.5 g glycerin 1 g Xylitol 0.5 g

2-6. Preparation of Granules

To the composition of Table 17, 100 mg of 30% ethanol was added and dried at 60 degrees Celsius to form granules, which were then filled into a cloth to prepare granules.

Furtherance Production Example  2-6 (mg) Diosmin 150 mg Soybean Extract 50 mg glucose 200 mg Starch 600 mg

[ Production Example  3]

Preparation of skin ointments

To the composition of Table 18, an external skin ointment containing diosmin was prepared.

Furtherance Production Example  3 ( weight% ) Diosmin 0.5 Diethyl sebacate 8.0 Prepayment 5.0 Polyoxyethylene oleyl ether phosphate 6.0 Sodium benzoate Quantity

Claims (11)

A cosmetic composition having a skin moisturizing improvement, skin exfoliation, skin elasticity improvement, erythema suppression, skin wrinkle improvement or skin photoaging improvement effect, containing diosmin or a cosmetically acceptable salt thereof as an active ingredient. The method of claim 1, The diosmin or its cosmetically acceptable salt is characterized in that it comprises 0.0001 to 20% by weight based on the total weight of the composition Cosmetic composition. The method of claim 1, The composition includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, essence, pack, mask pack, mask sheet, exfoliant, Soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, latex, press powder, loose powder and eye shadow Cosmetic composition. The method of claim 1, The composition is characterized in that it further comprises a skin wrinkle improvement component  Cosmetic composition. The method of claim 4, wherein The wrinkle improvement component further included is any one or more selected from the group consisting of vitamin C, retinoic acid, transforming growth factor (TGF), protein from animal placenta, betulinic acid and chlorella extract  Cosmetic composition. A health functional food composition having a skin moisturizing improvement, exfoliation, skin elasticity improvement, erythema suppression, skin wrinkle improvement or skin photoaging improvement effect, containing diosmin or a food acceptable salt thereof as an active ingredient. The method of claim 6, The composition is characterized in that it is prepared in any one formulation selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and rings. Health food composition. The method of claim 6, The composition is characterized in that it further comprises a skin wrinkle improvement component  Health food composition. The method of claim 8, The wrinkle improvement component further included is any one or more selected from the group consisting of vitamin C, retinoic acid, transforming growth factor (TGF), protein from animal placenta, betulinic acid and chlorella extract Health food composition. A quasi-drug composition having a skin moisturizing improvement, skin exfoliation, skin elasticity improvement, erythema suppression, wrinkle improvement or skin photoaging improvement effect, containing diosmin or a salt thereof as an active ingredient. Improve skin moisturization, exfoliate skin, enhance skin elasticity, inhibit erythema, improve wrinkles and / or skin, including administering or taking a composition containing diosmine or a pharmaceutically acceptable salt thereof as an active ingredient. How to improve photoaging

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