Anti-aging composition

Abstract

The present invention addresses the problem of providing an anti-aging external composition and an aging suppression method using the same. By using an anti-aging external composition containing a glutathione persulfide or a glutathione analogue persulfide, it is possible to suppress a decrease in cell proliferation ability and hyaluronic acid production ability after aging induction. In addition, the tyrosinase activity of melanoma cells induced by melanin can be inhibited, and pigmentation after spots, freckles and tanning can be improved by inhibiting melanin production.

Description

Technical Field

[0001] This invention relates to anti-aging compositions and methods for inhibiting aging. Background Technology

[0002] It is known that exposure to ultraviolet (UV) radiation causes photoaging, inflammation, and pigmentation. Aging itself is known to occur due to genetic and environmental factors, leading to a decrease in skin cell count and reduced hyaluronic acid production, resulting in decreased skin function, reduced hydration and elasticity, and the appearance of spots and wrinkles. Furthermore, it is known that spots, freckles, and post-sun pigmentation are caused by a significant increase in melanin production in the skin's pigment cells.

[0003] To date, as methods for inhibiting cellular aging, fibroblast activators with nicotinamide, hydrolysate of rice bran extract, and bamboo shoot extract as active ingredients have been reported (Patent Document 1), and compositions containing acacia tree extract for the prevention or treatment of photoaging, inflammation, pigmentation, and / or cancer caused by ultraviolet radiation have been reported (Patent Document 2). However, since acacia tree and bamboo shoots are natural products, there is a problem that the contained ingredients may vary depending on the time period.

[0004] In addition, as a method for inhibiting melanin production, formulations containing yeast extract and glutathione that inhibit melanin production, and cosmetics containing such formulations have been reported (Patent Document 3). Furthermore, as a sulfur-containing cosmetic, a cosmetic composed of levulinic acid and a sulfur compound has been reported (Patent Document 4). However, compounds such as reduced glutathione and sulfides have a sulfur-like odor, and oxidized glutathione has issues with potency. Therefore, there is a need for compositions with high potency, low odor, and the ability to inhibit melanin production.

[0005] Furthermore, a method for inhibiting aging by promoting glutathione production in organisms through the administration of polyamines has been reported (Patent Document 5). In recent years, persulfides, as glutathione-related substances, have attracted attention, and although their antioxidant activity is anticipated, no aging inhibition using persulfides has been reported to date.

[0006] Existing technical documents Patent documents Patent Document 1: Japanese Patent Application Publication No. 2022-138725 Patent Document 2: Japanese Patent Application Publication No. 2022-149769 Patent Document 3: Booklet No. WO2015 / 151867 Patent Document 4: Japanese Patent Application Publication No. 2002-326918 Patent Document 5: Japanese Patent Application Publication No. 2021-050180 Summary of the Invention The technical problem that the invention aims to solve The objective of this invention is to provide an anti-aging composition containing glutathione persulfate or glutathione analog persulfate, and a method for inhibiting aging using an anti-aging topical composition containing glutathione persulfate or glutathione analog persulfate.

[0007] Technical solutions for solving technical problems The inventors conducted in-depth research on the solutions to the above-mentioned problems and found that glutathione persulfate inhibits the aging of cells that have undergone aging-inducing treatment, thus completing the present invention.

[0008] That is, the present invention relates to the following first to sixth points.

[0009] First, an anti-aging composition containing 0.001 to 0.7% by weight of a persulfate of a glutathione analog or a persulfate of a cysteine ​​analog.

[0010] Second, an anti-aging composition, wherein the persulfate of the glutathione analog or the persulfate of the cysteine ​​analog is an oxidized glutathione trisulfide or an oxidized NAC trisulfide.

[0011] Third, an aging inhibition method, characterized in that the composition described in the first or second method is used as an oral or topical agent.

[0012] Fourth, according to the third method for inhibiting aging, wherein the method for inhibiting aging is a method for enhancing hyaluronic acid production or a method for inhibiting melanin production.

[0013] Fifth, a method for enhancing the inhibitory effect of glutathione analogs or cysteine ​​analogs on melanin production, characterized by persulfating the glutathione analogs or cysteine ​​analogs.

[0014] Sixth, a method for enhancing the hyaluronic acid production effect of glutathione analogues or cysteine ​​analogues, characterized in that the glutathione analogues or cysteine ​​analogues are persulfated.

[0015] Invention Effects According to the present invention, by using an anti-aging composition containing glutathione persulfate or glutathione analog persulfate, it is possible to inhibit the age-induced decrease in cell proliferation capacity and hyaluronic acid production of normal human fibroblasts and normal human epidermal keratinocytes, thereby improving skin wrinkles and enhancing skin elasticity and radiance. Attached Figure Description

[0016] Figure 1The figure shows the inhibition of aging-induced proliferation of normal human fibroblasts and the reduction in proliferation caused by persulfides.

[0017] Figure 2 The graph shows the inhibition of aging-induced proliferation of normal human epidermal keratinocytes and the reduction in proliferation caused by persulfides.

[0018] Figure 3 The graph shows the inhibition of hyaluronic acid production in normal human fibroblasts induced by aging and the reduction in hyaluronic acid production caused by persulfates.

[0019] Figure 4 This graph shows the ability of normal human epidermal keratinocytes to produce hyaluronic acid inhibited by aging and the reduced ability to produce hyaluronic acid caused by persulfates.

[0020] Figure 5 This is a graph showing the results of an experiment to inhibit melanin production.

[0021] Figure 6 This is a graph showing the results of a tyrosinase activity inhibition assay.

[0022] Figure 7 It is caused by persulfides in human skin. A graph showing the changes in values.

[0023] Figure 8 It is caused by persulfides in human skin. A graph showing the changes in values.

[0024] Figure 9 It is caused by persulfides in human skin. A graph showing the changes in values. Detailed Implementation

[0025] The compositions of the present invention comprise glutathione analog persulfates or cysteine ​​analog persulfates (hereinafter sometimes collectively referred to as persulfates). Here, glutathione analogs in the present invention refer to reduced or oxidized glutathione, reduced or oxidized γ-glutamylcysteine, reduced or oxidized cysteylglycine, and substances in which one or more amino or carboxyl groups in these structures are modified by a straight-chain or branched alkyl group having 1 to 24 carbon atoms, as well as substances esterified with organic acids or inorganic esters. Furthermore, cysteine ​​analogs in the present invention refer to cysteine, cystine, reduced or oxidized N-acetylcysteine ​​(hereinafter sometimes referred to as NAC), and substances in which one or more amino or carboxyl groups in these structures are modified by a straight-chain or branched alkyl group having 1 to 24 carbon atoms, as well as substances esterified with organic acids or inorganic esters. The persulfide in this invention refers to a molecule having a structure in which one or more sulfur atoms are directly bonded to the sulfur atom constituting the thiol group or disulfide bond in the molecules of general glutathione analogs or cysteine ​​analogs (e.g., RS-SH, RSSS-R'; R and R' are arbitrary functional groups). Any molecule having one or more sulfur atoms directly bonded to the sulfur atom in the glutathione analog or cysteine ​​analog molecule can be used in this invention, and can also be used as a mixture of compounds containing different numbers of sulfur atoms in various molecules.

[0026] The persulfide contained in the composition of the present invention is preferably an oxidized persulfide, more preferably an oxidized glutathione persulfide or an oxidized NAC persulfide, and even more preferably an oxidized glutathione persulfide (oxidized glutathione trisulfide) or an oxidized NAC persulfide (oxidized NAC trisulfide) containing 3 sulfur atoms in 1 molecule of persulfide.

[0027] Regarding the method for obtaining the persulfide of the present invention, it is possible to use the aforementioned glutathione analogs or cysteine ​​analogs that are commonly available as raw materials. For example, oxidized glutathione and peracetic acid are reacted on ice for 30 minutes, and after adding an equal amount of a solvent composed of mixed ethanol and THF (tetrahydrofuran), the mixture is centrifuged. The resulting precipitate is then further reacted with sodium hydrosulfide at room temperature, thereby obtaining oxidized glutathione persulfide. By purifying this oxidized glutathione persulfide, oxidized glutathione trisulfide can be produced, yielding substances in crystalline, amorphous, solution, or suspension states.

[0028] The composition of the present invention contains the above-mentioned persulfide as an active ingredient, and can also be used in the form of a mixture with other raw materials, provided that it does not impair the effect of the present invention. In addition to the above-mentioned persulfide, thickeners, pH adjusters, fragrances, bases for topical agents, etc., can also be mentioned.

[0029] Persulfides can be prepared by mixing them simultaneously with other raw materials during composition preparation, or by mixing them at different times. There are no restrictions on the form of the composition as long as it is suitable for oral ingestion or dermal application. It can be prepared using solvents and base materials commonly used in food, dietary supplements, cosmetic compositions, pharmaceutical compositions, and topical dermatological agents, and can be formulated into solutions, dispersions, powders, granules, capsules, tablets, pastes, gels, or sheets using conventional methods.

[0030] The content of each component in the composition of the present invention can be appropriately adjusted by adding excipients or the like commonly used as oral compositions or topical skin preparations.

[0031] The content of persulfides in the compositions of the present invention can be determined using common methods of HPLC. The content of persulfides in the compositions of the present invention can be appropriately adjusted, typically containing 0.001 to 0.7% by weight, preferably 0.0012 to 0.5% by weight.

[0032] In this invention, aging refers to the increase in wrinkles, decrease in elasticity and luster, decrease in the skin's ability to retain moisture, and the formation of spots and pigmentation. The anti-aging effect of this invention refers to the improvement of wrinkles, elasticity, luster, or brightness of the skin, the enhancement of the skin's ability to retain moisture (moisturizing), and the inhibition of pigmentation. It is believed that the amount of hyaluronic acid produced is related to the increase in wrinkles, decrease in elasticity and luster, and decrease in the skin's ability to retain moisture. Furthermore, it is believed that melanin production is related to the formation of spots and pigmentation. The degree of aging can be determined based on any commonly used aging indicator, such as cell proliferation capacity in cultured cells, decrease in hyaluronic acid production, melanin production, or in human skin, the increase in wrinkles, decrease in elasticity and luster, decrease in the skin's ability to retain moisture, or the formation of spots and pigmentation.

[0033] Furthermore, by using the extent to which the anti-aging indicators such as cell proliferation capacity, hyaluronic acid production capacity, or melanin production are improved as indicators, the anti-aging effect (aging inhibition effect) of the anti-aging composition can be determined.

[0034] For example, it is known that cell proliferation is reduced in aging cells, so it can be evaluated using common cell proliferation assays such as the MTT assay. Furthermore, it can be evaluated by the degree of inhibition of hyaluronic acid production or melanin production. Hyaluronic acid production capacity in aging cells can be quantitatively evaluated using common methods such as ELISA.

[0035] Furthermore, the composition of the present invention exhibits tyrosinase activity inhibition, thereby suppressing melanin production and inhibiting the formation of spots and pigmentation. The amount of melanin in cultured cells can be quantified using conventional methods, such as measuring absorbance at 405 nm. Alternatively, when measuring the amount of melanin in the skin, it can be measured and quantified using a melanin meter, for example. The reduction in the amount of melanin measured by any method indicates the effect of inhibiting melanin production.

[0036] Furthermore, the tyrosinase activity in this invention can be confirmed by adding a known concentration of L-DOPA to a protein extract containing tyrosinase from B16 mouse melanoma cells induced with melanin by melanin inducers such as α-MSH (melanocyte-stimulating hormone), and measuring the absorbance at 490 nm, correcting for the protein concentration in the protein extract. The tyrosinase activity inhibition effect can be determined by adding the test substance during melanin induction, and comparing the absorbance at 490 nm, corrected for protein concentration and measured using the same procedure, with the value in the test area without the test substance.

[0037] The above-mentioned composition for inhibiting melanin production contains persulfide as an active ingredient, and can therefore be used as a composition that inhibits melanin production by applying it directly to the skin or taking it orally.

[0038] Alternatively, as mentioned above, it can also be prepared into the desired dosage form for use.

[0039] The above-mentioned anti-aging composition contains persulfide as an active ingredient, and therefore can be used as an anti-aging composition by applying it directly to the skin as a topical agent.

[0040] Alternatively, as mentioned above, it can also be prepared into the desired dosage form for use.

[0041] Example The present invention will be specifically described below using examples, but the present invention is not limited to these examples.

[0042] The persulfide was prepared by the following method: Oxidized glutathione (Nacalai Tesque) was dissolved in ultrapure water to prepare a 170 mM (mol / L) aqueous solution. 4 mL of 32 wt% peracetic acid solution (Sigma-Aldrich) was added to 2.5 mL of this oxidized glutathione aqueous solution, and the mixture was reacted on ice for 30 min. An equal volume of a solvent containing a mixture of ethanol and THF (both from Nacalai Tesque) was added to the resulting reaction solution, and the mixture was centrifuged at 10,000 rpm for 10 min at 4 °C. The recovered precipitate was dried under reduced pressure for 1 hour. 2.5 mL of ultrapure water was added to the dried product for dissolution. An equal volume of a 60 mM sodium hydrosulfide solution prepared using 0.3 M NaOH was further added, and the mixture was reacted at room temperature for 1 hour to obtain the oxidized glutathione persulfide. The obtained oxidized glutathione persulfate was purified using a synthetic adsorption resin, concentrated, and then dried to obtain oxidized glutathione trisulfide powder (sample 1).

[0043] In addition, N-acetylcysteine ​​(Nacalai Tesque) was used instead of oxidized glutathione, and the same operation was performed to obtain powder of oxidized NAC trisulfide (sample 2).

[0044] (Inhibits the reduced ability of normal human fibroblasts to proliferate) Normal human fibroblasts were seeded into 96-well plates (3.0 × 10⁻⁶). 3 Cells / well, N=3), pre-cultured for 24 hours (5% CO2, 37°C). The culture medium used was MEM-α, nucleosides (Gibco) containing fetal bovine serum (10% added for serum-free medium). After washing the wells once with PBS(-), the medium was replaced with MEM-α, nucleoside medium containing fetal bovine serum (5% added for serum-free medium) dissolved at a final concentration of 50 μM or 100 μM as in Sample 1 or Sample 2, and incubated for another 30 minutes. Then, fibroblasts were treated with medium adjusted to a final concentration of hydrogen peroxide of 0.5 mM to induce senescence. MTT assays were then performed according to standard methods, and fibroblast viability was calculated as a relative value with the cell viability of the test area without any treatment or senescence induction set to 100%.

[0045] In addition, for comparison, a test area was set up where only aging induction was performed, and a test area was set up where 100 μM of reduced glutathione was used instead of sample 1.

[0046] For survival rates, significance tests (t-test, two-sided) were performed between the test areas of Sample 1 and Reduced Glutathione, and between the test areas of Sample 2 and Reduced Glutathione.

[0047] The results are shown in Figure 1 The test areas of Sample 1 and Sample 2 showed significantly higher cell proliferation rates compared to the test areas induced solely by aging and those induced by reduced glutathione (p < 0.005 for both). This demonstrates that oxidized glutathione trisulfide and oxidized NAC trisulfide can significantly inhibit the decrease in cell proliferation capacity induced by aging in normal human fibroblasts.

[0048] (Inhibits the reduction in the proliferative capacity of normal epidermal keratinocytes) Normal human epidermal keratinocytes were seeded into 96-well plates (1.0 × 10⁻⁶). 4 Cells / well, N=3), pre-cultured for 24 hours (5% CO2, 37℃). Commercially available keratinocyte proliferation medium (PromoCell) was used. After washing the wells once with PBS(-), the medium was replaced with keratinocyte proliferation medium dissolved to a final concentration of 50 μM or 100 μM (as in Sample 1), and incubated for another 30 minutes. Then, normal human epidermal keratinocytes were treated with medium adjusted to a final concentration of 0.5 mM hydrogen peroxide to induce aging. MTT assays were then performed according to standard methods, and the survival rate of normal human epidermal keratinocytes was calculated as a relative value with the cell survival rate of the test area (without oxidized glutathione trisulfide treatment or aging induction) set at 100%.

[0049] In addition, for comparison, a test area was set up where only aging induction was performed without oxidized glutathione trisulfide treatment.

[0050] For survival rate, significance tests (t-test, two-sided) were performed on the test area of ​​sample 1 and the test area that only underwent aging induction.

[0051] The results are shown in Figure 2 Compared to the test area induced by aging alone, the test area of ​​Sample 1 showed a significant increase in cell proliferation rate. This demonstrates that oxidized glutathione trisulfide can significantly inhibit the decrease in cell proliferation capacity induced by aging of normal human epidermal keratinocytes.

[0052] (Inhibits the decrease in hyaluronic acid production capacity induced by aging) Normal human fibroblasts were seeded into 24-well plates (8.0 × 10⁻⁶). 4Cells / well, N=3), pre-cultured for 24 hours (5% CO2, 37°C). The culture medium used was MEM-α, nucleoside (Gibco) containing fetal bovine serum (FBS) (10% added to serum-free medium). After washing the wells once with PBS(-), the medium was replaced with MEM-α, nucleoside medium containing FBS (5% added to serum-free medium) dissolved to a final concentration of 50 μM or 100 μM as in Sample 1 or Sample 2, and incubated for another 30 minutes. Then, fibroblasts were treated with medium adjusted to a final concentration of 0.5 mM hydrogen peroxide to induce senescence. After incubation for 24 hours with MEM-α, nucleoside medium containing FBS (5% added to serum-free medium), the hyaluronic acid level was quantified by ELISA using a hyaluronic acid quantification kit (Cosmo Bio).

[0053] The amount of hyaluronic acid produced in each test area is expressed as a relative value when the amount of hyaluronic acid produced in the test area without either the sample or the aging induction process is set to 100. Furthermore, for the amount of hyaluronic acid produced, significance tests (t-test, two-sided) were performed between the test area with sample 1 and the test area with only aging induction, and between the test area with sample 2 and the test area with only aging induction. Test areas showing significant differences were marked. .

[0054] The results are shown in Figure 3 It was found that the test areas of Sample 1 and Sample 2 had significantly higher hyaluronic acid levels compared to the test areas that underwent aging-induction only. Furthermore, the test areas of Sample 1 showed the same level of hyaluronic acid production capacity as the test areas that did not undergo aging-induction, with no significant difference identified between the two groups. The same results were observed for the test areas of Sample 2. Therefore, this suggests that oxidized glutathione trisulfide and oxidized NAC trisulfide can inhibit the decrease in hyaluronic acid production capacity induced by aging in normal human fibroblasts.

[0055] (Inhibits the decrease in hyaluronic acid production capacity induced by aging) Normal human epidermal keratinocytes were seeded into 24-well plates (1.0 × 10⁻⁶). 5Cells / well, N=3), pre-cultured for 24 hours (5% CO2, 37°C). Commercially available keratinocyte proliferation medium (PromoCell) was used. After washing the wells once with PBS(-), the medium was replaced with keratinocyte proliferation medium dissolved to a final concentration of 50 μM (sample 1), and incubated for another 30 minutes. Normal human epidermal keratinocytes were then treated with medium adjusted to a final concentration of 0.5 mM hydrogen peroxide to induce senescence. The medium was then replaced with fresh keratinocyte proliferation medium, and after incubation for 24 hours, the hyaluronic acid level was quantified by ELISA using a hyaluronic acid quantitative kit (Cosmo Bio). For comparison, an experimental area without oxidized glutathione trisulfide treatment was set up.

[0056] The amount of hyaluronic acid produced in each test area is expressed as a relative value when the amount of hyaluronic acid produced in the test area without oxidized glutathione trisulfide treatment and aging induction is set to 100. Furthermore, for the amount of hyaluronic acid produced, a significance test (t-test, two-sided) was performed between the test area of ​​Sample 1 and the test area with only aging induction. Test areas showing significant differences were marked. .

[0057] The results are shown in Figure 4 It can be seen that the test area of ​​sample 1 has a significantly higher amount of hyaluronic acid compared to the test area induced only by aging. Therefore, this suggests that it inhibits aging.

[0058] Furthermore, the test area of ​​sample 1 showed the same level of hyaluronic acid production capacity as the control area without aging induction, and no significant difference was identified between the two groups. This suggests that oxidized glutathione trisulfide can inhibit the decrease in hyaluronic acid production capacity induced by aging of normal human epidermal keratinocytes. Based on experiments to date, it is speculated that oxidized NAC trisulfide also has the same effect.

[0059] <Experiment on inhibiting melanin production in cultured cells> B16 mouse melanoma cells were seeded into 6-well plates (2.0 × 10⁶ cells per well). 4Cells (cells / well) were cultured in D-MEM medium supplemented with 5% fetal bovine serum for 24 hours (5% CO2, 37°C). Then, α-MSH (final concentration 200 nM) was added to serum-free D-MEM medium, and the cells were cultured for 24 hours to induce melanin production. After melanin induction, the cells were washed with PBS (-), treated with 0.25% trypsin (containing EDTA), and recovered. The cells were centrifuged at 10,000 rpm for 3 minutes to obtain particles. The obtained particles were washed twice with PBS (-), and 150 μL of 1M sodium hydroxide solution was added. The particles were then treated at 100°C for 10 minutes to dissolve them, thus preparing a reference sample.

[0060] In addition, during melanin induction, test areas were set for Sample 1 (final concentration 0.02mM or 0.05mM) or Sample 2 (final concentration 0.02mM or 0.05mM), and test areas for kojic acid (final concentration 1mM or 2.5mM) or reduced glutathione (final concentration 1mM or 3mM) as comparison objects, and samples were prepared.

[0061] Add 100 μL of sample from each test area to a 96-well plate and measure the absorbance at 405 nm using a microplate reader (n=3). Calculate the relative values ​​for each test area when the baseline sample's measurement value is set to 100. Perform a significance test on the baseline sample's measurement value using the Dunnett method. .

[0062] The results are shown in Figure 5 Comparison with the measured values ​​of the reference samples revealed significantly lower melanin production in the test areas of both Sample 1 and Sample 2 (p < 0.01). Furthermore, the 0.02 mM test area of ​​Sample 1 showed the same inhibitory effect on melanin production as the test area containing kojic acid, which is known to inhibit melanin production. However, the concentration of oxidized glutathione trisulfide was 0.02 mM, compared to 1 mM for the comparison compound, clearly demonstrating that oxidized glutathione trisulfide exhibited an inhibitory effect on melanin production at a lower concentration. Moreover, the test area containing reduced glutathione (final concentration 3 mM) also showed the same inhibitory effect on melanin production as the 0.02 mM test area of ​​Sample 1, but oxidized glutathione trisulfide showed an inhibitory effect on melanin production at a lower concentration. In the test area of ​​Sample 2, compared to the comparison compound, it also showed the ability to inhibit melanin production at a lower concentration. In conclusion, oxidized glutathione trisulfide exhibits superior inhibitory effects on melanin production in melanoma cells compared to reduced glutathione and kojic acid.

[0063] <Tyrosinase Activity Inhibition Assay> B16 mouse melanoma cells were seeded into 6-well plates (2.0 × 10⁶ cells per well).4 Cells / well were cultured in D-MEM medium supplemented with 5% fetal bovine serum for 24 hours (5% CO2, 37°C). Then, α-MSH (final concentration 200 nM) was added, and the cells were cultured in serum-free D-MEM medium for 24 hours to induce melanin production. After melanin induction, the cells were washed twice with PBS(-), treated with 0.25% trypsin (containing EDTA), and recovered. The cells were then centrifuged at 10,000 rpm for 3 minutes to obtain particles. The obtained particles were washed twice with PBS(-), lysed with 100 μL of lysis buffer (RIPABuffer; Fujifilm and Kagaku Purified Pharmaceuticals), and centrifuged at 4°C (14,000 rpm, 15 minutes). The supernatant was used as the sample for Comparative Example 5. Protein concentration was determined using the BCA method for the obtained samples.

[0064] Except for the absence of melanin induction, the same procedures were performed to prepare a reference sample. Additionally, during melanin induction, a test area was set up with sample 1 (final concentration 0.05 mM or 0.1 mM), and a test area for melanin induction only was used as a comparison. 50 μL of each test area sample was placed in a 96-well plate (n=3), and 50 μL of 0.8 mg / mL L-DOPA was added. After reacting at 37°C for 1 hour, the absorbance at 490 nm was measured using a microplate reader. The absorbance was corrected for protein concentration, and the relative values ​​of each test area were calculated when the reference sample's measurement value was set to 100. The Dunnett method was used to perform a significance test on the test areas for melanin induction only. .

[0065] The results are shown in Figure 6 The test areas supplemented with Sample 1 showed the same level of tyrosinase activity as the baseline sample at any concentration. Furthermore, compared to the test areas subjected to melanin induction alone, the tyrosinase activity was significantly lower. These results indicate that oxidized glutathione trisulfide can inhibit tyrosinase activity in melanoma cells.

[0066] The above indicates that glutathione analogue persulfate can inhibit tyrosinase activity and thus suppress melanin production in melanin-induced cultured cells. Furthermore, based on the results of the melanin-inhibiting experiments, it can be inferred that oxidized NAC trisulfide also exhibits the same effect.

[0067] <Whitening Effect Experiments on Humans> To confirm the whitening effect on humans, test lotions (lotions 1 and 2) containing the compositions of the present invention were prepared according to the composition in Table 1. Three participants who consented to the trial were selected as subjects. From November 10, 2023 to December 15, 2023, lotion 1 was applied twice daily (after bathing in the evening) to the inner side of the right forearm, and lotion 2 was applied to the test sites (5 sites each) on the inner side of the left forearm. Skin measurements were performed before and after the application period using the following method.

[0068] First, wash the test area with soap, dry it with paper towels, and allow it to acclimatize for 15 minutes. Then, using a colorimeter (SE-7700; Nippon Denshoku Kogyo Co., Ltd.) and optical fiber, measure the color at 5 points on the test area. value, Value and Calculate the average value. From the obtained value before coating... value, Value and The difference between the average values ​​after coating is calculated from the average values, and used as the mean. value, Value and Value. It should be noted that the uncoated areas were used as a control.

[0069] [Table 1] The results are shown in Figures 7-9 Compared to the uncoated area (control), The values ​​tend to be larger on areas where toners 1 and 2 were applied, suggesting improved skin radiance. It is speculated that these results contribute to an effect of inhibiting melanin production.

[0070] in addition, The value is a measure related to skin redness. The smaller the value, the more the redness is improved. According to... Figure 8 In areas where toners 1 and 2 were applied, smaller differences were observed compared to the control group. This is attributed to an increased ability to produce hyaluronic acid, resulting in improved skin texture and reduced redness.

[0071] at last, The value is related to items associated with dullness. The smaller the value, the more the dullness is improved. Figure 9 It can be seen that, compared with the control, the areas where toners 1 and 2 were applied showed smaller [size / weight / discoloration]. The dullness has improved.

[0072] In summary, it has been shown that applying toner 1 or 2 to the skin improves skin brightness and balances skin tone.

[0073] Based on the results of cell experiments, it is speculated that oxidized NAC trisulfide also shows the same effect.

[0074] As shown above, the composition of this invention can inhibit aging.

Claims

1. An anti-aging composition, characterized in that, Persulfides containing 0.001 to 0.7% by weight of glutathione analogs or cysteine ​​analogs.

2. An anti-aging composition, characterized in that, The persulfate of the glutathione analog or the persulfate of the cysteine ​​analog is an oxidized glutathione trisulfide or an oxidized NAC trisulfide.

3. An aging inhibition method, characterized in that, The composition according to claim 1 or 2 may be used as an oral or topical preparation.

4. The aging inhibition method according to claim 3, wherein, The aging inhibition method is either a method to enhance hyaluronic acid production or a method to inhibit melanin production.

5. A method for enhancing the inhibitory effect of glutathione analogues or cysteine ​​analogues on melanin production, characterized in that, Persulfation of glutathione analogues or cysteine ​​analogues.

6. A method for enhancing the hyaluronic acid production effect of glutathione analogues or cysteine ​​analogues, characterized in that, Persulfation of glutathione analogues or cysteine ​​analogues.

Images