Cosmetic composition containing nuezenide

Abstract

To provide a cosmetic composition with high beauty benefits. [Solution] A cosmetic composition containing nuezenid, preferably a composition derived from olive seeds.

Description

Technical Field

[0001] The present invention relates to a cosmetic composition, and more particularly to a composition containing nuezhenide and having cosmetic effects such as improving skin wrinkles.

Background Art

[0002] Plants contain various functional substances. In recent research, it has been clarified that plant-derived components have cosmetic effects such as improving skin functions. For example, an oral composition or a topical skin preparation containing fenugreek extract, which is effective in improving skin functions, has been proposed (see, for example, Patent Document 1).

Prior Art Documents

Patent Documents

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Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] An object of the present invention is to provide a novel cosmetic composition having high cosmetic effects.

Means for Solving the Problems

[0005] The present inventors have found that nuezhenide has excellent cosmetic effects such as improving skin wrinkles and skin elasticity, and have completed the present invention.

[0006] That is, the present invention is as follows. [1] A cosmetic composition characterized by containing nuezhenide. [2] The cosmetic composition according to [1] above, which is for improving skin wrinkles. [3] The cosmetic composition according to [1] above, which is for improving skin elasticity. [4] The composition according to any one of [1] to [3] above, characterized in that the nuezenid is derived from olive seeds.

[0007] [5] A cosmetic composition characterized by containing olive seeds or an extract thereof. [6] The cosmetic composition described in [5] above, characterized in that it is for improving wrinkles on the skin. [7] The cosmetic composition described in [5] above, characterized in that it is for improving the elasticity of the skin.

[0008] [8] A cosmetic composition characterized by containing olive oil residue or an extract thereof. [9] The cosmetic composition described in [8] above, characterized in that it is for improving wrinkles on the skin.

[10] The cosmetic composition described in [8] above, characterized in that it is for improving the elasticity of the skin. [Effects of the Invention]

[0009] The cosmetic composition of the present invention exhibits excellent cosmetic effects. [Brief explanation of the drawing]

[0010] [Figure 1] This figure shows the results of the evaluation of nuezenide's elastase inhibitory activity, and the graph shows the elastase activity of human dermal fibroblasts after the addition of nuezenide (n=3, mean ± SD, **p<0.01). [Figure 2] This figure shows the results of the evaluation of nuezenide's elastase inhibitory activity, and the graph shows the elastase inhibitory activity of human dermal fibroblasts after the addition of nuezenide (n=3, mean ± SD, **p<0.01). [Figure 3] This figure shows the results of the safety evaluation of nuezenide, and the graph shows the cell viability of human dermal fibroblasts after the addition of nuezenide (n=3, mean ± SD). [Figure 4] This graph shows the collagen production ratio of human dermal fibroblasts after the addition of nuezenide (n=3, mean ± SD, *p<0.05, **p<0.01). [Figure 5] Graph showing the ratio of collagen production per human skin fibroblast by adding nuezhenide (n = 3, mean ± SD). [Figure 6] Graph showing the change in R2, an index of the total elasticity of the skin, before and after using the nuezhenide-containing cream. [Figure 7] Graph showing the change in R7, an index of the recovery elasticity of the skin, before and after using the nuezhenide-containing cream.

Mode for Carrying Out the Invention

[0011] The beauty composition of the present invention is characterized by containing nuezhenide. The beauty composition of the present invention has an elastase inhibitory action and / or a collagen production promoting action. Therefore, the beauty composition of the present invention can improve skin wrinkles and / or improve skin elasticity by promoting the production of skin components such as elastin and collagen, and contribute to skin beauty.

[0012] That is, the beauty composition of the present invention can be used as a composition for inhibiting elastase, a composition for promoting collagen production, a composition for improving skin wrinkles, a composition for improving skin elasticity, and the like.

[0013] Here, the improvement of skin wrinkles in the present invention is a concept that includes not only reducing and eliminating existing wrinkles but also suppressing the occurrence of wrinkles. Also, the improvement of skin elasticity is a concept that includes not only improving (restoring) the firmness, flexibility, and elasticity of the skin but also suppressing the decrease thereof.

[0014] Hereinafter, each component contained in the beauty composition of the present invention will be described. Nuezhenide (Nuezhenide / Nuzhenide / Specnuezhenide) is a kind of iridoid glycoside and is, for example, a compound having the following chemical formula. Note that it may have substituents as long as the effects of the present invention are not inhibited.

[0015] [Chemistry]

[0016] As the nuezhenide contained in the cosmetic composition of the present invention, those derived from plants, those obtained by chemical synthesis, or commercially available reagents can be used. From the viewpoint of safety in oral ingestion, those derived from plants are preferred. Examples of plants containing nuezhenide include plants of the genus Olea in the family Oleaceae such as olive, and plants of the genus Ibotanoki in the family Oleaceae such as Tokunezumimochi. From the point that higher effects can be obtained, plants belonging to the genus Olea in the family Oleaceae are preferred, and Olea europaea is more preferred.

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[0057] The parts of the plant to which nuezenide can be applied are not particularly limited, as long as they contain nuezenide, including seeds, leaves, fruits (pulp and peel), stems, flowers, and roots.

[0020] In the case of olives, seeds are preferred. Furthermore, from the standpoint of effective resource utilization, it is preferable to use the residue (oil extraction residue) left after olive oil has been extracted. Normally, during oil extraction, the seeds and fruit are not separated and are extracted together. Therefore, from the standpoint of manufacturing efficiency, it is preferable to use the residue consisting of seeds and fruit. However, in cases where it is necessary to clearly define the extraction target (part of the plant), such as in cosmetics, it is preferable to use seed residue from which the fruit has been separated.

[0021] Examples of plant extracts include organic solvent extracts and aqueous organic solvent extracts. Examples of organic solvents include methanol, ethanol, isopropanol, butanol, ethyl acetate, methyl acetate, and acetone. Among these, ethanol extracts and aqueous ethanol extracts are preferred due to their safety and excellent cosmetic effects.

[0022] The method for obtaining the extract is not particularly limited and can be performed using commonly used methods (e.g., immersion extraction, reflux extraction, supercritical fluid extraction, etc.). Multiple extraction methods may also be combined. By combining multiple extraction methods, it is possible to obtain extracts with various compositions.

[0023] Furthermore, the extract may be used in liquid form or as a concentrate, but it is preferable to use it as a dried powder (extract powder). There are no particular restrictions on the drying method, and methods such as room temperature drying, heat drying, vacuum drying, and freeze-drying can be used.

[0024] Furthermore, it is preferable to use an extract that has been ultra-finely ground to about several tens of microns. The method for ultra-fine grinding the extract is not particularly limited, and known methods can be used. For example, ultra-fine grinding machines such as attritors, bead mills, and NGUs (nanogrinding units) can be used.

[0025] The cosmetic composition of the present invention can be used, for example, as a pharmaceutical product (including quasi-drugs), a cosmetic product, a functional food such as a food for specified health uses, a food with nutritional function claims, or a functional food whose efficacy has been approved by a designated institution, and other so-called health foods.

[0026] The composition of the present invention can be used as a topical or oral preparation. As for topical preparations, there are no particular restrictions as long as they are applied to the skin, etc. Examples of such preparations include ointments, creams, gels, lotions, emulsions, packs, patches, and other topical skin preparations, as well as injectable preparations.

[0027] Examples of oral preparations include tablets, capsules, powders, granules, liquids, granular preparations, sticks, plates, blocks, pills, pastes, creams, caplets, gels, chewables, and sticks. Among these, tablets, capsules, powders, granules, and liquids are particularly preferred.

[0028] The cosmetic composition of the present invention is not particularly limited as long as it contains nuezenide and can be distinguished from other products in terms of its use for skin beautification. For example, any product that displays a statement that it has skin beautifying effects on the product itself, packaging, instructions, or promotional materials (advertising media) is included within the scope of the present invention.

[0029] Furthermore, when using the cosmetic composition of the present invention as a composition for improving skin wrinkles or skin elasticity, such a composition is not particularly limited as long as it contains nuezenide and can be distinguished from other products in that it exhibits the function of improving skin wrinkles or skin elasticity. For example, any product that displays the function of improving skin wrinkles or skin elasticity on the product itself, packaging, instructions, or promotional materials (advertising media) is included in the scope of the present invention. In addition, products that display functions such as esterase inhibition or collagen production promotion as functions that contribute to improving skin wrinkles or skin elasticity are also included in the scope of the present invention.

[0030] For example, in the case of cosmetics and health foods, specific labels such as "skin beautification," "for those concerned about wrinkles," "prevents wrinkles and sagging," and "maintains skin firmness" can be displayed.

[0031] The amount of nuezenide contained in the cosmetic composition of the present invention should be appropriate, as long as nuezenide is present in a range that allows it to exert its effects as an active ingredient. For example, it is preferable to contain 0.001% by mass or more on a dry weight basis in the cosmetic composition of the present invention, more preferably 0.01 to 30% by mass, and even more preferably 0.05 to 10% by mass.

[0032] When the cosmetic composition of the present invention is an external preparation, there are no particular restrictions on the amount to be applied. However, from the viewpoint of more significantly exhibiting the effects of the present invention, it is preferable that the amount of nuezenide applied to an adult per day be 0.006 μg / day or more on a dry weight basis, more preferably 6 μg / day or more, even more preferably 18 μg / day or more, particularly preferably 25 μg / day or more, and most preferably 30 μg / day or more. The upper limit is, for example, 10 mg / day, preferably 5 mg / day.

[0033] When the cosmetic composition of the present invention is an oral preparation, there are no particular restrictions on the amount to be taken. However, from the viewpoint of more significantly exhibiting the effects of the present invention, it is preferable that the intake of nuezenide be 1 μg / day or more per adult, on a dry weight basis, more preferably 10 μg / day or more, and even more preferably 100 μg / day or more. The upper limit is, for example, 1000 mg / day, and preferably 100 mg / day.

[0034] The oral composition of the present invention can be contained in one container, or divided into, for example, two or three containers, to represent one day's intake, so that the daily intake amount is the aforementioned intake.

[0035] The composition of the present invention can be manufactured by known methods, with the addition of other components besides the components of the present invention as needed. Examples of other components besides the components of the present invention include nutritional components such as vitamins and minerals, and excipients. [Examples]

[0036] The present invention will be described below based on examples, but the present invention is not limited to these examples.

[0037] <Sample Preparation> As a sample, olive seeds (variety: Mission) were crushed to approximately 1 mm using a grinder, then further refined to approximately 40 μm using an NGU ultrafine grinder, and finally freeze-dried in a dryer to produce a frozen powder.

[0038] The freeze-dried powdered sample was immersed in 70% methanol for 60 minutes, then centrifuged at 3000 rpm for 15 minutes, and the supernatant was collected. This was repeated three times to obtain an olive concentrate. The obtained olive concentrate was fractionated by medium-pressure liquid chromatography (MPLC), and one of the fractions was isolated to obtain nuezenide.

[0039] <Elastase activity inhibition test> The dermis is mainly composed of collagen, elastin, and hyaluronic acid. Of these, elastin is produced by fibroblasts and plays an important role in maintaining skin flexibility and elasticity. The cross-linked structure formed by elastin and collagen is essential for maintaining skin elasticity. However, when elastase, an elastin-degrading enzyme, is induced by aging, UV radiation, reactive oxygen species, and stress, elastin is broken down, the cross-linked structure becomes brittle, skin elasticity is lost, and wrinkles form. In this study, the elastase inhibitory activity of nuezenid extract was measured to evaluate its function in restoring and maintaining skin elasticity.

[0040] In this study, the elastase inhibitory activity of nuezenide was evaluated by measuring the amount of p-nitroaniline produced using N-Succinyl-Ala-Ala-Ala-p-nitroanilide as a substrate in the presence of elastase derived from human dermal fibroblasts (NHDF-Ad) in human dermal fibroblasts (NHDF-Ad). Dimethyl sulfoxide (DMSO) was used as a control.

[0041] (Preparation of cell culture lysate) Human dermal fibroblasts (NHDF-Ad) were cultured in Dulbecco's Modified Eagle Medium (DMEM) (high glucose) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)) in a φ10 cm dish in a CO2 incubator (37°C, 5% CO2) until confluence. The cultured cells were washed with phosphate-buffered saline (PBS) and then lysed in cell lysate (0.2M Tris-HCl (pH 8.0) containing 1 mM PMSF and 5% Triton X-100) for 1.0 × 10⁻¹⁶ cells. 6 The cells were resuspended in cells / mL. After sonication, the cells were centrifuged (13,000 rpm, 4°C, 15 minutes), and the supernatant (celllysate) was collected.

[0042] (Measurement of elastase activity) To 12.5 μL of the recovered supernatant, 35.5 μL of 0.2 M Tris-HCl buffer (pH 8.0) and 2 μL each of nuezenide were mixed to the specified concentrations (final concentrations (μg / mL): 1, 10, 100, 200, 400, 1000, 2000), and the mixture was incubated at 37°C for 15 minutes.

[0043] Subsequently, 50 μL of 5 mM substrate (N-Succinyl-Ala-Ala-Ala-p-nitroanilide) was added to the test sample, and it was incubated at 37°C for 24 hours. Next, the absorbance of the obtained test sample at 405 nm was measured using a microplate reader, and the elastase activity of the test sample was calculated.

[0044] (Experimental results) The experimental results are shown in Figures 1 and 2. Figure 1 shows the elastase activity upon addition of nuezenide, with the elastase activity of the control sample set to 100% (control value), and Figure 2 shows the elastase inhibitory activity (%) based on the results of Figure 1. As shown in Figures 1 and 2, the addition of nuezenide resulted in a significant decrease in elastase activity (elastase inhibitory activity). Furthermore, the nuezenide extract exhibited concentration-dependent elastase inhibitory activity.

[0045] Based on these results, it was confirmed that nuezenide derived from olive seeds has elastase inhibitory activity, contributes to the restoration or maintenance of skin elasticity, and has an anti-wrinkle effect.

[0046] <Collagen production test> In this study, nuezenide was added to human dermal fibroblasts, and the changes in collagen production in the human dermal fibroblasts were measured to evaluate the collagen production-promoting effect of nuezenide, its anti-wrinkle effect, and its skin moisture-retaining effect.

[0047] (cell culture) Human dermal fibroblasts (NHDF-Ad) were used as a model cell. NHDF-Ad cells were pre-cultured in Dulbecco's Modified Eagle Medium (DMEM) (high glucose) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)) in φ10 cm dishes until confluent. Afterward, they were washed with phosphate-buffered saline (PBS), resuspended in culture medium, and then loaded into 96-well plates in 0.5 × 10⁶ wells. 4 Seeds were seeded at a concentration of cells / well and cultured overnight in a CO2 incubator (37°C, 5% CO2).

[0048] (added) After overnight incubation, the culture medium was replaced with serum-free DMEM medium (containing 1% penicillin-streptomycin) containing nuezenide (final concentrations of 0.1, 0.1, 1, 10, 100, 200, or 400 μg / mL), and incubated in a CO2 incubator for 72 hours.

[0049] (Measurement of cell viability) The cell viability in the culture medium 72 hours after the addition of each test substance was measured using CellCountingkit-8 (Dojin Chemical). 100 μL of DMEM (high glucose, containing 1% penicillin-streptomycin) was added to a 96-well plate from which the culture supernatant had been removed. Then, 3 μL of CCK-8 solution was added to each well, and the cells were incubated in a CO2 incubator for 1 hour. 90 μL of the cell supernatant was transferred to a 96-well plate, and the absorbance at 450 nm was measured using a microplate reader to calculate cell viability.

[0050] (Collagen production promotion evaluation test) The culture supernatant was collected 72 hours after the addition of each test substance, and the amount of collagen in the culture supernatant was measured using a commercially available ELISA kit (Human Collagen Type I ELISA, ACEL).

[0051] (Experimental results) As shown in Figure 3, NHDF-Ad cells treated with nuezenide showed a tendency towards increased cell viability compared to the control (DMSO). Furthermore, as shown in Figure 4, collagen production also significantly increased or showed an increasing trend compared to the control. Additionally, as shown in Figure 5, collagen production per cell showed a concentration-dependent increasing trend.

[0052] Based on these results, it was confirmed that nuezenide contributes to cell viability and collagen production.

[0053] <Clinical Trials> The olive-derived ingredient "nuezenid" was investigated through a clinical trial involving healthy individuals to determine its effectiveness in maintaining and improving skin elasticity.

[0054] (Preparation of test samples) An extract containing nuezenide was extracted from olive seeds using an aqueous alcohol (50% butylene glycol: BG) extraction solvent. A nuezenide-containing cream (test cream) was prepared using this nuezenide-containing olive seed extract. The amount of olive seed extract in the test cream was 10%. The test cream was applied to the face twice a day, with a total amount of cream used per day of 1.2g, and the amount of active ingredient (nuezenide) contained was 0.0324mg / day.

[0055] (test) Participants were instructed to discontinue the use of wrinkle-reducing cosmetics for one week prior to the intervention as a reset period. During the 8-week intervention period, participants applied a designated cream to their face twice a day. Before and after the intervention, the viscoelasticity of the cheeks was measured using a skin viscoelasticity measuring device (Cutometer DUAL MPA580 (Courage+Khazaka)). There were 8 participants in the study.

[0056] (result) As shown in Figures 6 and 7, R2, an indicator of total skin elasticity, and R7, an indicator of recovery elasticity, significantly increased before and after the use of the test cream. The significant improvement observed in these two viscoelasticity indicators in clinical trials on human skin suggests that the viscoelasticity of the skin was maintained or improved by suppressing the breakdown of components such as collagen and elastin that make up the dermis.

[0057] [Formulation Example 1] A cream formulation for cosmetic use was manufactured by combining the following ingredients. Olive seed extract powder Glycerin ethanol

[0058] [Formulation Example 2] A gel formulation for cosmetic use was manufactured by combining the following ingredients. Olive seed extract powder olive oil Ester oil

[0059] [Formulation Example 3] A powder formulation for capsules was manufactured by combining the following ingredients. Olive (fruit and seed) residue powder Calcium stearate

[0060] [Formulation Example 4] The tablets were manufactured by combining the following ingredients. Olive (fruit and seed) residue powder starch Dextrin [Industrial applicability]

[0061] The composition of the present invention is industrially useful because it can be used in cosmetics, health foods, and the like.

Claims

1. A cosmetic composition characterized by containing nuezenide.

2. The cosmetic composition according to claim 1, characterized in that it is for improving wrinkles on the skin.

3. The cosmetic composition according to claim 1, characterized in that it is for improving skin elasticity.

4. The cosmetic composition according to any one of claims 1 to 3, characterized in that the nuezenid is derived from olive seeds.

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