Agent and composition for suppressing darkening of skin or preventing formation of wrinkles

Abstract

Provided is an agent comprising a silver skin as a functional material. The agent can be used for purposes pertaining to beautifying effects. The agent contains a silver skin-derived extract as an active ingredient, and is configured for suppressing darkening of the skin or prevention of wrinkle formation. As the silver skin extract contains an elastase inhibitor, the agent is used for prevention of wrinkle formation. As the silver skin extract contains at least any one of a glycation suppressing agent, a protein carbonylation suppressing agent, a protein nitration suppressing agent, and a nitrotyrosine decomposing agent, the agent is used for suppression of darkening.

Description

Agents and compositions for suppressing skin dullness and preventing wrinkle formation

[0001] The present invention relates to the effective use of silver skin, and in particular to an agent for at least suppressing dullness of the skin and preventing wrinkle formation.

[0002] Silverskin (often found in large quantities in coffee roasting factories) refers to the thin skin that encases the coffee bean. Botanically, the coffee cherry is a drupe, and as shown in Figure 6, the coffee cherry is arranged in the following order from the outside to the center: husk, pulp, mucilage, parchment (endocarp), silverskin, and seed. The seeds have an embryo and endosperm, and are arranged facing each other within a single coffee cherry. As the coffee cherry matures, the flat surfaces of the endosperm on opposite sides are depressed and folded into the interior of the endosperm, forming a groove called a center cut in the center of this flat surface.

[0003] Green beans are those from which the outer part of the seed has been removed, and the process of extracting green beans from the coffee cherry is called refining. During the refining process of green coffee beans, some of the silverskin is removed, and most of the remaining silverskin falls off during roasting, typically resulting in industrial waste. Traditionally, silverskin has been partially used as a material for psicose production (see Patent Document 1), paper production (see Patent Document 2), company envelopes, business cards, New Year's cards, wrapping paper, toys, and other uses. It has also been known that silverskin can be effectively used as a functional material, such as a hyaluronidase inhibitor (see Patent Document 3 and Non-Patent Document 1).

[0004] Japanese Patent Application Laid-Open No. 2004-143062 Japanese Patent Application Laid-Open No. 2008-255519 Japanese Patent No. 5537924

[0005] Kazuya Iwai et al., “Inhibitory Effect of a Hot Water Extract of Coffee “Silverskin” on Hyaluronidas”, Biosci. Biotechnol. Biochem., Vol.75(6), pp.1205-1207, 2011.

[0006] The present inventors have conducted extensive research to more effectively utilize silverskin, which has traditionally been treated primarily as industrial waste. As a result, they have discovered that, in addition to being used as a hyaluronidase inhibitor, silverskin can also be used to suppress skin dullness and prevent wrinkle formation. Silverskin is derived from natural products, is highly safe for humans, and can be applied or ingested on a daily basis. In view of the above circumstances, the present invention aims to provide an agent that can utilize silverskin as a functional material for cosmetic purposes.

[0007] The agent of the present invention is an agent for at least suppressing skin dullness and preventing wrinkle formation, containing an extract derived from silver skin as an active ingredient. That is, the agent of the present invention can be used as an agent for suppressing skin dullness, an agent for preventing wrinkle formation, or an agent for suppressing skin dullness and preventing wrinkle formation. As used herein, "silver skin" refers to the thin skin surrounding the endosperm of Rubiaceae plants (such as the genus Coffea or Mascalocoffea). For example, the thin skin enveloping coffee beans is generated in large quantities in coffee roasting factories. Silver skin may be obtained from either green coffee beans or roasted coffee beans. Components of coffee bean silver skin itself include caffeine, 5-hydroxymethylfurfural, chlorogenic acid, and the like. The caffeine content of roasted coffee bean silver skin ranges from approximately 0.8 to 1.4 g / 100 g. Coffee bean silverskin is composed mostly of dietary fiber (60-80%), including cellulose, hemicellulose, and lignin. The main monosaccharides contained in coffee bean silverskin are xylose, arabinose, rhamnose, fucose, mannose, glucose, galactose, and uronic acid (a derivative obtained by oxidizing monosaccharides). The protein content of coffee bean silverskin is approximately 18%, while the content of other nutrients, such as fat and reducing sugars, is low. The fat content of silverskin is approximately 7.5 g / 100 g, and the fatty acids are primarily saturated (64%), followed by polyunsaturated (30%) and monounsaturated (6%). Coffee bean silverskin also contains phenolic compounds, which have high antioxidant properties.

[0008] Here, the agent of the present invention contains an elastase inhibitor in the silver skin extract and can be used to prevent wrinkle formation. This agent has cosmetic effects such as wrinkle prevention, improving skin elasticity, and anti-aging. From another perspective, the silver skin-derived extract can be used as an elastase inhibitor.

[0009] Furthermore, the agent of the present invention contains a silver skin extract that contains a glycation inhibitor and can be used to suppress dullness. This agent has cosmetic effects such as suppressing skin dullness, improving skin elasticity, and anti-aging. From another perspective, the silver skin-derived extract can be used as a glycation inhibitor.

[0010] Furthermore, the agent of the present invention contains a silver skin extract that contains a protein carbonylation inhibitor and can be used to suppress dullness. Protein carbonylation is a protein denaturation that causes a decrease in skin transparency. By suppressing carbonylation, this agent has cosmetic effects such as suppressing skin dullness, improving skin elasticity, and anti-aging. From another perspective, the silver skin-derived extract can be used as a protein carbonylation inhibitor.

[0011] Furthermore, the agent of the present invention contains a silver skin extract that contains a protein nitration inhibitor and can be used to suppress dullness. Protein nitration causes yellowing and dullness of the skin. This agent suppresses nitration, and therefore has cosmetic effects such as suppressing dullness of the skin, improving skin elasticity, and anti-aging. From another perspective, the silver skin-derived extract can be used as a protein nitration inhibitor.

[0012] Furthermore, the agent of the present invention contains a silver skin extract and a nitrotyrosine decomposing agent, and can be used to suppress dullness. Nitrotyrosine (3-nitrotyrosine) is a type of nitrated protein modification, and is produced by the nitration of tyrosine residues by reactive nitrogen species such as nitric oxide, nitrogen dioxide, and peroxynitrite. Nitrotyrosine causes yellowing and dullness of the skin, and this agent decomposes nitrotyrosine, thereby suppressing dullness of the skin, improving skin elasticity, and providing cosmetic effects such as anti-aging. From another perspective, an extract derived from silver skin can be used as a nitrotyrosine decomposing agent.

[0013] The agent of the present invention preferably contains an extractant in an amount 10 times or more and 20 times or less the amount of the silver skin-derived extract. Furthermore, the agent of the present invention preferably contains an extract derived from silver skin of 1000 ppm (0.1%) or more. In this specification, the base units of % and ppm are mass. The agent of the present invention can be used as an anti-aging agent. It is effective for skin beautification and beauty, suppressing dullness of the skin and preventing wrinkle formation, and can be used as an anti-aging agent that can maintain a younger and healthier appearance than the actual age. Furthermore, this agent has the effect of reducing the risk of disease and extending healthy lifespan.

[0014] The agent of the present invention is a cosmetic used as a cosmetic composition for suppressing skin dullness and preventing wrinkle formation, a food used as a food composition, a drug used as a pharmaceutical composition, or a quasi-drug used as a quasi-drug composition for use in preventing skin dullness and wrinkle formation. In addition, in any of the methods for suppressing skin dullness and preventing wrinkle formation of the present invention, the agent of the present invention is used as an oral preparation or a skin application preparation.

[0015] According to the present invention, silver skin can be effectively used as a functional material, which is highly safe for humans and is useful for suppressing dullness of the skin and preventing wrinkle formation.

[0016] Graph comparing glycation inhibition rates Graph comparing carbonylation inhibition rates Graph comparing nitration inhibition rates Graph comparing nitrated tyrosine decomposition rates Graph comparing elastase activity inhibition rates Silver Skin diagram

[0017] (Silver Skin Extraction Procedure) An extract was prepared from silver skin obtained from roasted beans, and its effect on skin efficacy was evaluated. The silver skin was weighed into three beakers, and 10 times, 20 times, and 30 times the amount of distilled water was added. The mixture was then heated at 60°C for 4 hours and filtered using filter paper and a membrane filter to obtain an extract. The extract was concentrated using an evaporator and then freeze-dried to obtain a dried product (extract).

[0018] The extraction results are shown in Table 1. When the amount of solvent used was small (10 times the amount), most of the added solvent was absorbed by the silverskin, and the extract could not be obtained without squeezing it out. The results shown in Table 1 (1) are the results of squeezing, but the extract yield was low at 43%. On the other hand, as shown in Table 1 (2), when the amount of solvent was 20 times or more, there were no problems with operability, similar to obtaining extracts from other common plants, and the extract yield was 66.5%. Furthermore, as shown in Table 1 (3), the extract yield also improved with increasing the amount of solvent (43% → 66.5% → 78%), but the higher the extract yield, the lower the solid content in the extract (1.53% → 0.77% → 0.54%). Therefore, for subsequent tests, the extract (dried product) obtained by extraction with 20 times the amount of distilled water shown in Table 1 (2) was used.

[0019]

[0020] As described above, when obtaining an extract from silverskin, if the ratio of solvent to silverskin (raw material) is low (10 times the amount), the yield of the extract is low (less than 50%), which poses problems in terms of operability. However, it was found that an extract can be obtained without problems if the amount of extraction solvent is 20 times or more. Because silverskin is highly water-absorbent, if the amount of water is 10 times the amount (10 times the amount), the silverskin absorbs the water, making extraction difficult without squeezing. Therefore, solid-liquid separation by natural settling (decantation) or gravity filtration, etc., has problems in terms of extraction time and yield. On the other hand, even when the amount of water is 10 times the amount (10 times the amount), squeezing, centrifugation, vacuum filtration, or other squeezing operations rather than solid-liquid separation can shorten the extraction time and improve the yield. Therefore, although an extract can be obtained without problems if the amount of extraction solvent is 20 times or more, if the amount is more than 20 times the amount, the concentration process becomes costly and time-consuming, so it is preferable to perform extraction at a low water addition ratio.

[0021] (Glycation Inhibition Test) The results of evaluating the presence or absence of glycation inhibitory activity of silver skin extract are shown below. Glycation in the skin is involved in various skin aging processes, and it is known that the glycation of skin proteins and the accumulation of advanced glycation end products (AGEs) due to aging and glycation stress cause the skin to yellow and become dull. Proteins such as collagen and elastin, which are the main components of the dermis, have long half-lives and are susceptible to glycation, so glycation of proteins in the dermis is involved in maintaining the stability of fibrous tissue and can also cause a decrease in firmness and elasticity. For this reason, inhibiting glycation is effective in maintaining skin.

[0022] The test sample was silverskin extract, and aminoguanidine hydrochloride was used as a positive control. The silverskin extract was dissolved in distilled water to prepare sample solutions so that the final concentrations of the silverskin extract in the test system could be adjusted to 100 ppm and 1000 ppm. The final concentration of aminoguanidine hydrochloride (positive control) was 40 mM.

[0023] For the test method, D-glucose, D-ribose, and bovine serum albumin were dissolved in 1 M phosphate buffer to a final concentration of 10%, 1%, and 1%, respectively. The sample solution was added to this solution to give final concentrations of silver skin extract of 100 ppm and 1000 ppm, and the mixture was then incubated at 60°C for 3 days. For the control, phosphate buffer was added instead of the sample. After the reaction, 200 μL of the reaction solution was dispensed into a 96-well plate, and the fluorescence intensity at 465 nm when excited at 360 nm was measured using a fluorescence microplate reader. The amount of advanced glycation end products produced by the reaction was calculated, and the glycation inhibition rate at each concentration of silver skin extract was calculated based on the amount of advanced glycation end products in the control.

[0024] The results of the glycation inhibition test are shown in Figure 1. When silverskin (CS: Coffee Silverskin) extract was added at 100 ppm and 1000 ppm, no glycation inhibition effect was observed at 100 ppm, but an inhibitory effect was observed at 1000 ppm. These results confirmed that silverskin extract has an inhibitory effect on glycation at a certain concentration or higher.

[0025] (Carbonylation Inhibition Test) Next, the results of an evaluation of the carbonylation inhibitory effect of silver skin extract are shown. Aldehydes such as acrolein, a decomposition product of lipid peroxide, add to proteins, carbonylating them, and it is known that carbonylated proteins are present in large amounts in the stratum corneum, epidermis, and dermis tissues of sun-exposed skin. Carbonylation in the skin has been linked to factors such as the yellowing of dermal proteins due to carbonylation, a decrease in optical transparency due to carbonylated proteins in the sun-exposed stratum corneum, and dry skin due to carbonylation of keratin in the stratum corneum. Therefore, inhibiting carbonylation in the skin is effective in maintaining healthy skin.

[0026] The test sample was silverskin extract, and aminoguanidine hydrochloride was used as a positive control. The silverskin extract was dissolved in distilled water to prepare sample solutions so that the final concentrations of the silverskin extract in the test system could be adjusted to 100 ppm and 1000 ppm. The final concentration of aminoguanidine hydrochloride was 40 mM.

[0027] For the test method, a 96-well plate was prepared by coating with Cellmatrix® Type 1-A collagen. 100 μL of sample solution was dispensed into each well to achieve final concentrations of 100 ppm and 1000 ppm of 0.5 mM acrolein and silver skin extract. Distilled water was dispensed into control wells. The n number for each sample was six. After overnight reaction at 37°C, the reaction solution was removed, washed with distilled water, and stained at room temperature with 200 μL of 20 μM fluorescein-5-thiosemicarbazide / 0.1 M MES-Na (pH 5.5). After staining, the staining solution was removed, and the plate was thoroughly washed with T-PBS. The fluorescence intensity at 535 nm when excited at 465 nm was measured using a fluorescence microplate reader. The amount of carbonylation due to the reaction was calculated, and the carbonylation inhibition rate at each concentration of silver skin extract was calculated based on the amount of carbonylation in the control.

[0028] The results of the carbonylation inhibition test are shown in Figure 2. When silverskin (CS) extract was added at 100 ppm and 1000 ppm, no carbonylation inhibition effect was observed at 100 ppm, but a weak carbonylation inhibition effect was observed at 1000 ppm. These results confirm that silverskin extract has a carbonylation inhibition effect at a certain concentration or higher.

[0029] (Nitration Inhibition Test) The results of an evaluation of silver skin extract for its inhibitory effect on nitration are shown below. Nitration is a post-translational protein modification in which reactive nitrogen species generated in vivo add a nitro group to the benzene ring of tyrosine or tryptophan residues that make up proteins. Protein nitration affects cellular function by causing a decrease in the function of enzymes and tyrosine kinase receptors, and the accumulation of nitrotyrosine in proteins is associated with numerous diseases, including arteriosclerosis and ischemic cerebral disease. Protein nitration is also known to be involved in a variety of diseases and symptoms in various tissues within the body. Furthermore, it has been suggested that protein nitration contributes to a yellowish discoloration of the skin, and that in the stratum corneum, nitration of stratum corneum cell adhesion factors is associated with poor stratum corneum desquamation. For these reasons, preventing nitration in the skin is crucial.

[0030] The test sample was silverskin extract, and ferulic acid was used as a positive control. The silverskin extract was dissolved in distilled water to prepare sample solutions with final concentrations of 100 ppm and 1000 ppm in the test system. The final concentration of ferulic acid was 3 mM.

[0031] For the test method, tyrosine solution was dispensed into 1.5 mL tubes, and the sample solution was added to this solution and mixed so that the final concentrations of silverskin extract were 100 ppm and 1000 ppm. Distilled water was added instead of the sample to the control. Peroxynitrite was added to a concentration of 100 μM, and the mixture was incubated at 37°C for 24 hours. The reaction solution was then filtered, and the amount of nitrotyrosine produced by the reaction was quantified using HPLC. The amount of nitrotyrosine produced by the reaction was calculated, and the nitration inhibition rate at each concentration of silverskin extract was calculated based on the amount of nitrotyrosine in the control.

[0032] The results of the nitration inhibition test are shown in Figure 3. When silverskin (CS) extract was added at 100 ppm and 1000 ppm, almost no nitration inhibitory effect was observed at 100 ppm, but a strong nitration inhibitory effect was confirmed at 1000 ppm. These results confirm that silverskin extract has a strong nitration inhibitory effect at a certain concentration or higher.

[0033] (Regarding the Nitrated Product Decomposition Test) The following shows the results of an evaluation of the decomposition activity of nitrated products (here, nitrotyrosine) in silver skin extract. It has been shown that nitration has adverse effects in the skin, and it is known that the cause of yellowing and dullness of the skin is not only the glycosylated and carbonylated proteins present in the dermis, but also nitrated proteins formed by the nitration of amino acids that make up proteins present in the stratum corneum. Therefore, if the nitrated proteins formed by nitration can be decomposed, various symptoms caused by protein nitration, such as yellowing and dullness of the skin, can be expected to improve.

[0034] The test sample was silverskin extract, and sodium dithionite was used as a positive control. The silverskin extract was dissolved in distilled water to prepare sample solutions so that the final concentrations of silverskin extract in the test system could be adjusted to 100 ppm and 1000 ppm. The final concentration of sodium dithionite was 1 mM.

[0035] For the test method, 25 μL of sample solution was added to each well of a 96-well plate so that the final concentrations of silverskin extract were 100 ppm and 1000 ppm, and 225 μL of 0.005% nitrotyrosine aqueous solution was then dispensed. Distilled water was added instead of the sample to the control. The 96-well plate was sealed and incubated at 37°C for 72 hours. After the reaction, the absorbance at 450 nm was measured using a microplate reader. The ratio of the amount of nitrotyrosine when silverskin extract was added to the amount of nitrotyrosine in the control was calculated, and the percentage of nitrotyrosine reduction was calculated as the nitrotyrosine decomposition rate.

[0036] The results of the nitrate decomposition test are shown in Figure 4. When silverskin (CS) extract was added at 100 ppm and 1000 ppm, no decomposition of nitrotyrosine was observed at 100 ppm, but the decomposition of nitrotyrosine was confirmed at 1000 ppm. These results confirm that silverskin extract has the ability to decompose nitrates at a certain concentration or higher.

[0037] (Elastase Activity Inhibition Test) The results of an evaluation of silver skin extract to determine whether it has the ability to inhibit the enzymatic activity of elastase, an elastin-degrading enzyme, are shown below. It is well known that wrinkles are formed through long-term exposure to ultraviolet light, i.e., photoaging, and it is generally accepted that changes in the dermal matrix are an important factor. It is also known that photoaging changes in elastin, an elastic fiber that is one of the dermal matrix components, in skin through an increase in normal fibers and thickening of fibers, leading to the formation of amorphous masses. On the other hand, it is also known that the activity of elastase, an elastin-degrading enzyme, significantly increases in skin exposed to UVA. For this reason, it is important to suppress the elastase activity that increases after ultraviolet exposure as a means of preventing wrinkles caused by photoaging.

[0038] The test sample was silverskin extract, and ethylenediaminetetraacetic acid (EDTA) was used as a positive control. The silverskin extract was dissolved in distilled water to prepare sample solutions with final concentrations of 100 ppm and 1000 ppm in the test system. The final concentration of ethylenediaminetetraacetic acid was 250 μM.

[0039] For the test method, 8 mM elastase substrate (Methoxy-succinyl-alanyl-alanyl-propyl-valine-p-nitroanilide) was dispensed into each well of a 96-well plate, and sample solution was dispensed so that the final concentrations of silver skin extract were 100 ppm and 1000 ppm. Distilled water was added instead of the sample to the control. 0.25 units / mL human leukocyte elastase was then added, mixed, and incubated at 37°C for 20 minutes. Absorbance at 405 nm was then measured using a microplate reader. The amount of substrate decomposition by elastase in the control and silver skin extract treatments was calculated based on the absorbance of the light-absorbing substance produced by decomposition of the elastase substrate. Furthermore, the ratio of the amount of substrate decomposition in the silver skin extract treatment to the amount of elastase substrate decomposition in the control was calculated, and the percentage reduction in substrate decomposition was calculated as the elastase activity inhibition rate.

[0040] The results of the elastase activity inhibition test are shown in Figure 5. When silverskin (CS) extract was added at 100 ppm and 1000 ppm, almost no elastase activity inhibitory effect was observed at 100 ppm, but an elastase activity inhibitory effect was confirmed at 1000 ppm. These results confirm that silverskin extract has an elastase activity inhibitory effect at a certain concentration or higher.

[0041] (Cosmetic compositions, food compositions, pharmaceutical compositions, quasi-drug compositions) 1) Cosmetic compositions can be selected appropriately depending on the purpose, and examples thereof include lotions, lotions, creams, and emulsions. 2) Food compositions can be selected appropriately depending on the purpose, and examples thereof include beverages such as coffee, tea, soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks, as well as foods such as candy, jelly, gum, and chocolate. 3) Pharmaceutical compositions can be selected appropriately depending on the purpose, and examples thereof include tablets, capsules, energy drinks, and lozenges. 4) Quasi-drug compositions can be selected appropriately depending on the purpose, and examples thereof include medicated soaps and bath additives.

[0042] The present invention is useful in cosmetic compositions and food compositions for suppressing dullness of the skin and preventing wrinkle formation.

Claims

1. A skin dullness suppressant with an active ingredient derived from silver skin.

2. The skin dullness suppressant according to claim 1, wherein the extract contains a glycation inhibitor and is used to suppress skin dullness.

3. The skin dullness suppressant according to claim 1, wherein the extract contains a protein carbonylation inhibitor and is used to suppress skin dullness.

4. The skin dullness suppressant according to claim 1, wherein the extract contains a protein nitration suppressant and is used to suppress skin dullness.

5. The skin dullness suppressant according to claim 1, wherein the extract contains a nitrotyrosine decomposing agent and is used for suppressing skin dullness.

6. A glycation inhibitor that uses an extract derived from silver skin as an active ingredient to prevent dullness of the skin.

7. A protein carbonylation inhibitor that uses an extract derived from silver skin as an active ingredient to prevent skin dullness.

8. A protein nitration inhibitor that uses an extract derived from silver skin as an active ingredient to prevent dullness of the skin.

9. A nitrotyrosine decomposing agent that uses an extract derived from silver skin as an active ingredient to prevent dullness of the skin.

10. The agent according to any one of claims 1 to 9, wherein the amount of extraction solvent in the extract is 10 times or more and 20 times or less.

11. The agent according to any one of claims 1 to 9, wherein the content of the extract is 1000 ppm or more.

12. 10. The agent according to any one of claims 1 to 9, which is used as an anti-aging agent.

13. A cosmetic product comprising the agent according to any one of claims 1 to 9 as a cosmetic composition.

14. A food product comprising the agent according to any one of claims 1 to 9 as a food composition.

15. A pharmaceutical product comprising the agent according to any one of claims 1 to 9 as a pharmaceutical composition.

16. A quasi-drug in which the agent according to any one of claims 1 to 9 is used as a quasi-drug composition.

17. A method for suppressing dullness of the skin, comprising administering the agent according to any one of claims 1 to 9 as an oral preparation.

18. A method for suppressing dullness of the skin, comprising using the agent according to any one of claims 1 to 9 as a skin application agent.