Novel composition

Abstract

To provide a novel composition that aids improvement such as skin moisturization, and a novel promoter for aquaporin expression.SOLUTION: The composition of the present invention (for example, a topical skin preparation such as a cosmetic) contains glyceryl glucoside, pomegranate extract, and lemongrass extract. In the total amount of the composition, the combined content of glyceryl glucoside, pomegranate extract, and lemongrass extract is 0.00001 mass% or more and less than 5.0 mass%, and of these three components, glyceryl glucoside has the highest content.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to a novel composition, and in particular to a composition that can promote the expression of aquaporins and increase the moisturizing power of the skin, and is useful in cosmetics and the like. [Background technology]

[0002] Maintaining skin moisture, elasticity, and barrier function is important for maintaining healthy, youthful skin. In recent years, with the growing trend toward anti-aging, many cosmetics focused on these functions have been sold.

[0003] Aquaporins are proteins with pores present in cell membranes, known as water channels that selectively allow water molecules to pass through. The activity of these water channels increases the cell membrane's water permeability. Thirteen types of aquaporins (0-12) are known to exist in humans, with aquaporin 3 being particularly prevalent in the skin. For example, it has been reported that mice lacking aquaporin 3 exhibit significantly reduced skin moisture and elasticity, as well as delayed wound healing. Thus, it is believed that enhancing aquaporin production can improve skin moisture, elasticity, and roughness.

[0004] It has been reported that various naturally occurring ingredients and substances promote the expression of aquaporins. For example, Patent Document 1 reports that the expression level of aquaporin 3 is increased by adding an extract such as lemongrass. Patent Document 2 lists pomegranate extract as an aquaporin-stimulating compound that stimulates aquaporin membrane proteins in the epidermis. It has also been reported that glyceryl glucoside, known as a moisturizing agent, has the ability to induce the expression of aquaporin 3 mRNA in keratinocytes.

[0005] However, there remains a need for compositions that contribute to improving skin moisturization and the like. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Laid-Open No. 2013-087057 [Patent Document 2] Special Publication No. 2009-500394 Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a novel composition that contributes to improving skin moisture, etc. Another object of the present invention is to provide a novel aquaporin expression promoter. [Means for solving the problem]

[0008] As a result of extensive research aimed at solving the above-mentioned problems, the present inventors have found that a combination of glyceryl glucoside, pomegranate extract, and lemongrass extract exhibits a synergistic effect of promoting aquaporin expression compared to when each extract is used alone.

[0009] The composition of the present invention is characterized by containing glyceryl glucoside, pomegranate extract, and lemongrass extract. The total content of the glyceryl glucoside, pomegranate extract, and lemongrass extract in the composition is 0.00001% by mass or more and less than 5.0% by mass. The composition is further characterized by having the highest content of the glyceryl glucoside among the glyceryl glucoside, pomegranate extract, and lemongrass extract.

[0010] The aquaporin expression promoter of the present invention is characterized by containing glyceryl glucoside, pomegranate extract, and lemongrass extract, and the aquaporin is aquaporin 3. [Effects of the Invention]

[0011] The present invention can provide a composition (for example, a cosmetic) that exhibits a good aquaporin expression promoting effect and contributes to improving functions such as moisturizing of the skin. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 shows the results of aquaporin 3 gene expression levels when each component was used alone. [Figure 2] FIG. 1 shows the results of cell viability of epidermal keratinocytes when each component is used alone. [Figure 3] FIG. 1 shows the results of aquaporin 3 gene expression levels when ingredients are used in combination. [Figure 4] FIG. 1 shows the results of cell viability of epidermal keratinocytes when ingredients are used in combination. DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention provides a novel composition comprising (a) glyceryl glucoside, (b) pomegranate extract, and (c) lemongrass extract.

[0014] (a) Glyceryl glucoside Glyceryl glucoside is a sugar derivative having a structure in which glucose and glycerin are glycosidicly bonded. Depending on the glycerin bond site and the steric configuration of glucose, there are 1-α-glyceryl glucoside, 1-β-glyceryl glucoside, 2-α-glyceryl glucoside, and 2-β-glyceryl glucoside. In the present invention, any of these may be used, or a mixture of two or more of these may be used.

[0015] Glyceryl glucoside can be produced by chemical synthesis, microbial fermentation, etc. Glyceryl glucoside may be used as a single raw material, may be used as a mixed raw material with other components (various solvents, etc.), or may be a commercially available product.

[0016] The content of glyceryl glucoside in the composition of the present invention is preferably 0.00001% by mass or more, more preferably 0.0001% by mass or more, and may be 0.0005% by mass or more, based on the total amount of the composition (100% by mass). The content of glyceryl glucoside is preferably less than 5.0% by mass based on the total amount of the composition. The content of glyceryl glucoside in the present invention is the amount converted into a pure content (when used as a mixed raw material, the amount excluding other components).

[0017] (b) Pomegranate extract Pomegranate (scientific name: Punica granatum) is a deciduous tree of the genus Punica in the family Lythraceae. Pomegranate flowers around June and bears fruit in autumn. A characteristic feature of pomegranate fruit is that the skin splits open irregularly when ripe. In the present invention, "pomegranate extract" refers to an extract obtained by extracting the pomegranate plant (fruit, skin, etc.), and preferably refers to an extract obtained by extracting pomegranate fruit.

[0018] The extraction process for pomegranate extract is not particularly limited, and may be, for example, solvent extraction using water or an organic solvent. Examples of organic solvents include monohydric alcohols such as ethanol, propanol, and isopropanol, polyhydric alcohols such as 1,3-butylene glycol and glycerin, acetone, and ethyl acetate. The organic solvent may be used alone or in combination with water. For example, when using an aqueous ethanol solution, a 10% to 95% v / v ethanol solution is used.

[0019] The form of the pomegranate plant to be subjected to the extraction treatment is not particularly limited, and for example, dried or crushed pomegranate fruit or fresh pomegranate fruit may be used. In addition, the extraction temperature, amount of extraction solvent used, extraction time, etc. in the extraction treatment can be appropriately set.

[0020] The pomegranate extract may be used as it is, or may be filtered and concentrated as necessary, or may be fractionated and purified using column chromatography or the like.

[0021] The pomegranate extract may be used as a single ingredient, or may be used as a mixed ingredient with other ingredients (various solvents, etc.), or a commercially available product may be used.

[0022] The content of pomegranate extract in the composition of the present invention is preferably 0.000001% by mass or more, more preferably 0.00001% by mass or more, based on the total amount of the composition. Furthermore, the content of pomegranate extract is preferably less than 5.0% by mass, based on the total amount of the composition. The content of pomegranate extract in the present invention is the amount of solids excluding the solvent.

[0023] (c) Lemongrass extract Lemongrass (scientific name: Cymbopogon citratus) is a perennial plant of the Poaceae family, belonging to the Cymbopogon genus. Lemongrass has a scent similar to lemon and is used to flavor tea and cooking. In the present invention, "lemongrass extract" refers to an extract obtained by extracting the leaves and / or stems of lemongrass.

[0024] The extraction process for lemongrass extract is not particularly limited, and may be, for example, solvent extraction using water or an organic solvent. For example, the extraction solvents described in (b) above can be used. The lemongrass extract may be used as is as an extract, or may be filtered and concentrated as necessary. It may also be fractionated and purified using column chromatography or the like.

[0025] The lemongrass extract may be used as a single raw material, or may be used as a mixed raw material with other components (various solvents, etc.), or a commercially available product may be used.

[0026] The content of lemongrass extract in the composition of the present invention is preferably 0.000001% by mass or more, more preferably 0.000005% by mass or more, based on the total amount of the composition. Furthermore, the content of lemongrass extract is preferably less than 3.0% by mass, based on the total amount of the composition. The content of lemongrass extract in the present invention is the amount of solids excluding the solvent.

[0027] The total content of the above (a) to (c) in the total amount of the composition of the present invention is, for example, 0.00001 mass% or more and less than 5.0 mass%, preferably 0.0001 mass% or more and less than 3.0 mass%, and may be 0.0005 mass% or more and less than 3.0 mass%.

[0028] In the composition of the present invention, among the above (a) to (c), it is preferable that the content of (a) glyceryl glucoside be the highest, and more preferably the order is (glyceryl glucoside) > (pomegranate extract) > (lemongrass extract). This makes it easier to effectively exert the effect of promoting aquaporin gene expression while further considering safety. In this case, for example, if the content (mass%) of lemongrass extract is "1," the content ratio of glyceryl glucoside is preferably 5 to 200, and the content ratio of pomegranate extract is preferably 2 to 100. Alternatively, the content ratio of glyceryl glucoside may be 20 to 200, and the content ratio of pomegranate extract may be 2 to 15.

[0029] As shown in the Examples below, the composition of the present invention has the effect of promoting the expression of aquaporin genes. Aquaporins are proteins with pores present in cell membranes. They are water channels that selectively allow water molecules to pass through but do not allow ions or other substances to pass through. They form tetramers within the cell membrane, and each monomer functions as a water channel. The composition of the present invention has the effect of promoting the expression of aquaporin 3 genes, which are particularly expressed in the skin.

[0030] The composition of the present invention is preferably a topical skin preparation to be applied to the skin. The topical skin preparation can be various cosmetics, quasi-drugs, or pharmaceuticals. For example, the cosmetic form can be skin care cosmetics such as lotions, emulsions, or creams, or makeup cosmetics. Furthermore, the quasi-drug or pharmaceutical form can be an ointment, cream, liquid, or the like.

[0031] The composition of the present invention may contain other moisturizing ingredients, various oils, surfactants, ultraviolet absorbers, powders, pigments, polyhydric alcohols, thickeners, antioxidants, fragrances, preservatives, and the like.

[0032] Other moisturizing ingredients may be any moisturizing ingredient commonly used in the field of topical skin preparations. Examples include polyhydric alcohols such as glycerin, polyethylene glycol, propylene glycol, 1,3-butylene glycol, dipropylene glycol, and pentylene glycol; sugars such as sorbitol, glycosyltrehalose-hydrogenated starch hydrolyzate mixture, maltitol, trehalose, and raffinose; amino acids such as valine, leucine, alanine, arginine, glutamine, lysine, aspartic acid, and glutamic acid; betaine; and plant extracts such as Sagarame extract and carrot root extract. These moisturizing ingredients may be used alone or in combination. Among the above moisturizing ingredients, it is preferable to include at least one moisturizing ingredient selected from glycerin and 1,3-butylene glycol.

[0033] Examples of oils that can be used include hydrocarbon oils such as liquid paraffin and squalane, ester oils such as isopropyl palmitate, vegetable oils such as soybean oil, olive oil, and castor oil, silicone oils such as methylpolysiloxane, higher fatty acids such as oleic acid, higher alcohols such as stearyl alcohol and behenyl alcohol, alkyl glyceryl ethers such as batyl alcohol, petrolatum, lanolin, and shea butter. One or more types of oils can be used.

[0034] The surfactant may be an ionic surfactant or a nonionic surfactant. Examples of nonionic surfactants that can be used include glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, propylene glycol fatty acid esters, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, and polyoxyethylene hydrogenated castor oil. One or more surfactants may be used.

[0035] The aquaporin expression promoter of the present invention promotes the expression of aquaporin 3, which is mainly localized in the skin, thereby improving water permeability in the skin and contributing to the prevention and improvement of symptoms and phenomena caused by a decrease in skin moisture content, including skin aging such as wrinkle formation and loss of elasticity. Therefore, the aquaporin expression promoter of the present invention is used for the purpose of promoting the expression of aquaporin genes. It is particularly preferably used for the purpose of promoting the expression of aquaporin 3 genes. [Example]

[0036] [Test A: Evaluation of single use] In Test A, commercially available products were used for (a) glyceryl glucoside, (b) pomegranate extract, and (c) lemongrass extract. Pomegranate fruit extract was used for (b) pomegranate extract. Each commercially available product was dissolved in water or an alcoholic solvent, and the concentrations shown below are values ​​converted to pure products.

[0037] [Test A-1: ​​Evaluation test for promoting aquaporin 3 gene expression] The expression level of the aquaporin 3 gene was measured for the medium (Med) group, control group (Ctrl), and treatment groups at each concentration using the following procedure. Note that for Ctrl, the solvent contained in each commercially available product was adjusted to have a concentration similar to that of the treatment group.

[0038] Aquaporin 3 expression levels were examined using keratinocytes derived from normal human neonatal epidermis. Specifically, keratinocytes were cultured until 50-70% confluent, then replaced with working medium and cultured in a 37°C, 5% CO2 incubator. After culture, total RNA was extracted using an RNA recovery reagent, and cDNA was synthesized from 2.5 μg of RNA using reverse transcriptase. Gene expression comparisons were performed using a real-time PCR system, with the ΔΔCT method using SYBR Green to measure the expression levels of the above genes. GAPDH was used as an internal standard.

[0039] The results of aquaporin 3 gene expression levels when used alone are shown in Figure 1. As shown in Figure 1, when (a) glyceryl glucoside, (b) pomegranate extract, and (c) lemongrass extract were used alone, a tendency to promote aquaporin 3 gene expression in a concentration-dependent manner was confirmed. In particular, a significant increase in aquaporin 3 gene expression was confirmed in the high concentration group (21.6 mg / ml) of (a) glyceryl glucoside.

[0040] [Test A-2: Cytotoxicity test (MTT test)] Cytotoxicity was evaluated at the test concentrations evaluated in Test A-1 above, according to the following procedure. Epidermal keratinocytes were seeded at 10,000 cells / well in a 12-well plate. The medium was changed the next day, and continued to be changed as needed until the cells became subconfluent. The medium was removed, and 1 ml / well of treatment medium adjusted to each concentration was added and incubated for 48 hours. After 48 hours, the treatment medium was removed, and 1 ml / well of MTT-containing medium was added and incubated for 4 hours. After 4 hours, 1 ml / well of isopropanol (containing 0.04 N HCl) was added and the reaction was stopped by stirring for 15 minutes on a plate mixer. 200 μL / well of the stopped solution was transferred to a 96-well plate, and the absorbance (570 nm - 630 nm) was measured using a microplate reader. The cell viability was calculated using the following formula. Cell viability (%) = (absorbance of each well - absorbance of blank) / (absorbance of control - absorbance of blank) x 100

[0041] The results of cytotoxicity when used alone are shown in Figure 2. As shown in Figure 2, when a 48-hour MTT test was performed, the cell viability was 80% or higher in all treatment groups compared to the Ctrl group, and no significant toxicity was observed. Glyceryl glucoside (a), which showed a particularly significant effect in the above test A-1, had a higher concentration than the other components (b) and (c), but no difference in cell viability was observed.

[0042] [Test B: Evaluation of combined use] In Test B, as in Test A, commercially available products were used for (a) glyceryl glucoside, (b) pomegranate extract, and (c) lemongrass extract.

[0043] [Test B-1: Evaluation test for promoting aquaporin 3 gene expression] The test was carried out in the same manner as in Test A-1 above. Regarding the concentrations of (a) to (c), the concentrations of each component in Test A were set at low concentrations, taking into consideration the cytotoxicity when used in combination. Specifically, the medium used was supplemented with (a) glyceryl glucoside at 2.7 mg / ml, (b) pomegranate extract at 0.15 mg / ml, and (c) lemongrass extract at 0.002625%. In this case, the contents of each component in the medium were in the following order: (glyceryl glucoside) > (pomegranate extract) > (lemongrass extract).

[0044] The results of the expression level of the aquaporin 3 gene when the components were used in combination are shown in Figure 3 and Table 1. In Table 1, "+" indicates the addition of each component, and "-" indicates that each component was not added. As shown in Table 1, Comparative Examples 1 to 3 are cases where a single component was used, Comparative Examples 4 to 6 are cases where two components were used in combination, and Example 1 is a case where three components were used in combination.

[0045] [Table 1]

[0046] As shown in Figure 3 and Table 1, Example 1, which used the three ingredients (a) to (c) in combination, significantly increased the expression level of the aquaporin 3 gene, significantly promoting the expression of the aquaporin 3 gene. In contrast, when the ingredients were used alone or in combination, promoting the expression of the aquaporin 3 gene was observed, but to a lesser extent. Furthermore, simply using each ingredient in combination does not necessarily improve the effect; for example, Comparative Example 5, which used (a) glyceryl glucoside and (c) lemongrass extract in combination, resulted in a decrease in the expression level of the aquaporin 3 gene compared to Comparative Example 3, which used (c) lemongrass extract alone. Thus, surprisingly, Example 1 of the present invention was confirmed to have a synergistic effect on promoting aquaporin expression.

[0047] [Test B-2: Cytotoxicity test (MTT test)] Cytotoxicity was evaluated at the test concentrations evaluated in the above Test B-1. The test procedures were the same as those in the above Test A-2.

[0048] The results of the cytotoxicity of the combined use are shown in Figure 4. As shown in Figure 4, when a 48-hour MTT test was performed, even in Example 1, in which the three components (a) to (c) were used in combination, the cell viability was 80% or more compared to the Ctrl group, and no significant toxicity was observed. Therefore, it is possible to achieve a significant effect in promoting aquaporin expression while ensuring safety.

[0049] From the above, it was found that the combination of (a) glyceryl glucoside, (b) pomegranate extract, and (c) lemongrass extract synergistically promotes the expression of aquaporins. The composition and aquaporin expression promoter of the present invention are particularly useful for topical skin preparations (such as cosmetics) that are administered to the skin. Furthermore, each of the above components (a) to (c) has a proven track record of use in topical skin preparations, which also helps ensure safety.

Claims

1. A composition comprising glyceryl glucoside, pomegranate extract and lemongrass extract.

2. The composition according to claim 1, characterized in that the total content of the glyceryl glucoside, the pomegranate extract and the lemongrass extract is 0.00001% by mass or more and less than 5.0% by mass of the total amount of the composition.

3. 3. The composition according to claim 2, wherein the content of glyceryl glucoside is the highest among the glyceryl glucoside, the pomegranate extract, and the lemongrass extract in the total amount of the composition.

4. 3. The composition according to claim 1, wherein the composition is a cosmetic.

5. An aquaporin expression promoter characterized by containing glyceryl glucoside, pomegranate extract, and lemongrass extract.

6. The aquaporin expression promoter according to claim 5, wherein the aquaporin is aquaporin 3.

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