Cosmetic composition for skin improvement, comprising nicotinamide adenine dinucleotide

Abstract

The present invention relates to a cosmetic composition for skin improvement, comprising nicotinamide adenine dinucleotide, and more particularly, to a cosmetic composition in which intracellular potency of nicotinamide adenine dinucleotide is enhanced so as to increase a concentration of nicotinamide adenine dinucleotide delivered into skin cells, thereby providing effects of inhibiting skin aging, improving skin texture, alleviating wrinkles, improving skin elasticity, promoting skin regeneration, and strengthening the skin barrier.

Description

Cosmetic composition for skin improvement containing nicotinamide adenine dinucleotide

[0001] The present invention relates to a cosmetic composition for improving skin containing nicotinamide adenine dinucleotide, and more specifically, to a cosmetic composition containing a combination of nicotinamide adenine dinucleotide, glycyrrhetinic acid, and quercetin, which increases the amount of nicotinamide adenine dinucleotide delivered into skin cells and provides a skin aging inhibitory effect.

[0002]

[0003] Nicotinamide adenine dinucleotide is highly biochemically unstable. Therefore, to achieve noticeable skin-improving effects, it must be incorporated into cosmetics in excessive amounts. Even then, its effectiveness is limited by homeostasis mechanisms.

[0004] Nicotinamide adenine dinucleotide phosphate (NADP), a derivative of nicotinamide adenine dinucleotide, is an alternative to nicotinamide adenine dinucleotide to address the intracellular potency issue. However, it is unclear whether this derivative has the same effect of inhibiting skin aging as nicotinamide adenine dinucleotide.

[0005] Against this backdrop, the inventors of the present invention have completed the present invention by confirming that by preparing a cosmetic composition using a combination of nicotinamide adenine dinucleotide, glycyrrhetinic acid, and quercetin to solve the problem of intracellular potency of nicotinamide adenine dinucleotide, the problem of conventional potency is solved, stability is secured, and the amount of nicotinamide adenine dinucleotide delivered into skin cells when applied to the skin is not reduced by various mechanisms, and a skin aging inhibition effect is provided.

[0006]

[0007] The present invention aims to provide a cosmetic composition that improves the potency of exogenous nicotinamide adenine dinucleotide.

[0008]

[0009] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. In other words, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below.

[0010]

[0011] A first aspect of the present invention for achieving the aforementioned purpose is to provide a cosmetic composition comprising nicotinamide adenine dinucleotide; glycyrrhetinic acid; and quercetin.

[0012] A second aspect of the present invention for achieving the aforementioned purpose is to provide a cosmetic composition comprising nicotinamide adenine dinucleotide; glycyrrhetinic acid; quercetin; and sitosterol or tocopheryl acetate.

[0013]

[0014] Hereinafter, the present invention will be described in more detail.

[0015]

[0016] The term "nicotinamide adenine dinucleotide (NAD+)" of the present invention is based on nicotinamide, a type of vitamin B3, and functions as an electron carrier in biochemical reactions such as oxidation and reduction within cells. It also plays an important role in preventing skin aging by generating skin cell energy, alleviating stress, repairing DNA, and controlling inflammation. As we age, the concentration of NAD+ within skin cells naturally decreases, which weakens the skin's natural regenerative ability and accelerates aging. Therefore, methods for maintaining or increasing NAD+ levels within skin cells are being studied as an important strategy for alleviating skin aging.

[0017] As mentioned above, due to its highly biochemically indeterminate properties, NAD+ is rarely directly incorporated into cosmetic compositions. Furthermore, NAD+ is rapidly converted to an inactive form within cells, making it difficult to expect sufficient anti-aging and wrinkle-improving effects when applied to the skin.

[0018] Accordingly, the present invention sought to solve the above problem by using glycyrrhetinic acid, quercetin, and sitosterol or tocopheryl acetate together to maintain a high concentration and bioactivity within cells when NAD+ is applied to the skin.

[0019]

[0020] The term "glycyrrhetinic acid" used in the present invention is one of the main active ingredients of licorice, possessing skin-soothing, anti-inflammatory, antioxidant, and antibacterial properties. It is also included in cosmetic compositions to help maintain healthy skin by reducing skin irritation and protecting it from external irritants.

[0021] The nicotinamide adenine dinucleotide and glycyrrhetinic acid may be included in a weight ratio of 1: 0.0004 to 1: 0.4, 1: 0.0004 to 1: 0.3, 1: 0.0004 to 1: 0.25, 1: 0.004 to 1: 0.4, 1: 0.004 to 1: 0.3, 1: 0.004 to 1: 0.25, 1: 0.01 to 1: 0.4, 1: 0.01 to 1: 0.3 or 1: 0.01 to 1: 0.25.

[0022]

[0023] The term "quercetin" used in the present invention is a polyphenol compound of the flavonoid family, found naturally in various plants, including fruits, vegetables, leaves, seeds, and grains. It is a powerful antioxidant and is known to possess diverse physiological activities, including anti-inflammatory, antibacterial, anti-allergy, and anti-cancer properties. It is particularly effective in preventing skin aging, restoring damaged skin, and relieving irritation and inflammation.

[0024] The above nicotinamide adenine dinucleotide quercetin may be included in a weight ratio of 1: 0.0004 to 1: 0.4, 1: 0.0004 to 1: 0.3, 1: 0.0004 to 1: 0.25, 1: 0.004 to 1: 0.4, 1: 0.004 to 1: 0.3, 1: 0.004 to 1: 0.25, 1: 0.01 to 1: 0.4, 1: 0.01 to 1: 0.3 or 1: 0.01 to 1: 0.25.

[0025]

[0026] The term "sitosterol" used in the present invention refers to a type of plant sterol, a steroidal substance derived from nature. It is primarily extracted from vegetable oils such as soybeans, wheat, corn, peanuts, and sesame seeds. In particular, it protects and soothes the skin through its moisturizing, anti-inflammatory, antioxidant, and emulsifying properties, and provides a restorative effect on damaged skin.

[0027] The nicotinamide adenine dinucleotide and sitosterol may be included in a weight ratio of 1: 0.0004 to 1: 0.4, 1: 0.0004 to 1: 0.3, 1: 0.0004 to 1: 0.25, 1: 0.004 to 1: 0.4, 1: 0.004 to 1: 0.3, 1: 0.004 to 1: 0.25, 1: 0.01 to 1: 0.4, 1: 0.01 to 1: 0.3 or 1: 0.01 to 1: 0.25.

[0028]

[0029] The term "tocopheryl acetate" in the present invention is a derivative of vitamin E, a synthetic compound combined with acetic acid. Vitamin E (tocopherol), known as a powerful antioxidant, is sensitive to light and heat and easily decomposes in cosmetics. However, tocopheryl acetate is more stable under these conditions. Tocopheryl acetate provides antioxidant, skin protection, regeneration, moisturizing, and skin-improving effects.

[0030] The nicotinamide adenine dinucleotide and tocopheryl acetate may be included in a weight ratio of 1: 0.0004 to 1: 1, 1: 0.0004 to 1: 0.8, 1: 0.0004 to 1: 0.5, 1: 0.004 to 1: 1, 1: 0.004 to 1: 0.8, 1: 0.004 to 1: 0.5, 1: 0.01 to 1: 1, 1: 0.01 to 1: 0.8, or 1: 0.01 to 1: 0.5.

[0031]

[0032] NAD+ is known to activate biological mechanisms within cells related to cellular lifespan extension and longevity (Laurent Mouchiroud, Cell, 2013). NAD+ is required for the activation of sirtuin proteins, and increasing intracellular NAD+ has been shown to have anti-aging effects.

[0033] However, when NAD+ is injected into skin cells from outside, it is rapidly degraded by their own homeostasis mechanisms. The most representative mechanism is NAD+ degradation by the CD38 protein, also known as NADase (NAD+ degrading enzyme). It has been shown that inhibiting CD38 activity or expression can increase the concentration of intracellular NAD+ (Carlos Escande, Diabetes. 2013).

[0034] In the present invention, it was confirmed that when skin cells are treated with NAD+, CD38 is overexpressed, and that quercetin, glycyrrhetinic acid, sitosterol, tocopheryl acetate, and combinations thereof effectively suppress this CD38 overexpression. Through this, it was confirmed that the concentration of NAD+ within skin cells can be effectively increased, i.e., the bioactivity of exogenous NAD+ can be significantly increased.

[0035]

[0036] The composition of the present invention may be used for inhibiting skin aging or photoaging.

[0037]

[0038] In another aspect, the present invention provides a method for inhibiting skin aging or photoaging, comprising the step of applying a cosmetic composition comprising nicotinamide adenine dinucleotide; glycyrrhetinic acid; and quercetin to the skin of a subject.

[0039] In the present invention, the term “subject” may mean any mammal, including humans, in need of anti-aging, photoaging inhibition, wrinkle or skin regeneration improvement.

[0040] In the present invention, the term “application” means contacting or providing the composition according to the present invention to the skin of a subject, and may be applied using a general route that reaches the skin or is absorbed into the skin.

[0041]

[0042] The cosmetic composition (cosmetics) of the present invention comprises all kinds of ingredients usable in cosmetics, such as moisturizers such as glycerin, butylene glycol, propylene glycol, hexanediol, methylgluceth-20, diglycerin, and ethylhexylglycerin; sunscreens such as ethylhexyl methoxycinnamate, ethylhexyl salicylate, ethylhexyl triazone, octocrylene, and bis-ethylhexyloxyphenol methoxyphenyl triazine; pH adjusters such as triethanolamine; thickeners such as carbomer, xanthan gum, acrylates / C10-30 alkyl acrylate crosspolymer, and hyaluronic acid; preservatives such as phenoxyethanol, methylparaben, and propylparaben; antioxidants such as BHT, ethyl ascorbyl ether, and ascorbic acid; skin conditioning agents such as beta-glucan; Surfactants such as cetearyl glucoside and sorbitan stearate; may additionally include fragrances or colors, but there is no limitation on the ingredients.

[0043] Each of the above-mentioned ingredients included in the cosmetic according to the present invention may be included in the cosmetic composition of the present invention within a range that preferably does not exceed the maximum usage amount stipulated in the “Cosmetics Safety and Technology Standards” established by each government.

[0044] The cosmetic according to the present invention can be manufactured in any formulation commonly manufactured in the art. For example, the cosmetic can have the formulation of, but is not limited to, a toner such as a flexible toner or a nourishing toner, a spray-type toner, an emulsion such as a facial lotion or a body lotion, a cream such as a nourishing cream, a moisturizing cream, an eye cream, a stick, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation such as a liquid type or a spray type, a makeup remover such as a powder, a cleansing lotion, a cleansing oil, a cleanser such as a cleansing foam, a soap, a body wash, etc.

[0045] In one specific example, the formulation of the cosmetic may be a balm, a water-in-oil (W / O) formulation, an oil-in-water (O / W) formulation, a solubilized formulation, or an oil formulation.

[0046] The cosmetic of the present invention can be used according to a conventional method of use, and the number of times it is used can vary depending on the user's skin condition or preference.

[0047]

[0048] In a specific embodiment, it was confirmed that the cosmetic composition of the present invention increases the concentration of nicotinamide adenine dinucleotide in skin cells and provides anti-aging, skin texture improvement, wrinkle improvement, skin regeneration, and barrier strengthening effects.

[0049]

[0050] The advantages and features of the present invention, and the methods for achieving them, will become clearer with reference to the experimental examples and manufacturing examples described in detail below. However, the present invention is not limited to the experimental examples and manufacturing examples disclosed below, but may be implemented in various different forms. These examples are provided solely to ensure the complete disclosure of the present invention and to fully inform those skilled in the art of the invention of the scope of the invention.

[0051]

[0052] The cosmetic composition of the present invention, when applied to the skin, increases the concentration of nicotinamide adenine dinucleotide in skin cells, and provides anti-aging, skin texture improvement, wrinkle improvement, skin regeneration, and barrier strengthening effects.

[0053]

[0054] Figure 1 is a photograph showing the cell aging results of Experimental Example 3 of the present invention.

[0055] Figure 2 is a graph showing the results of the cell replication life extension effect of Experimental Example 5 of the present invention.

[0056]

[0057] Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention more specifically, and the scope of the present invention is not limited by these examples.

[0058]

[0059] Experimental Example 1: Confirmation of CD 38 expression inhibition ability

[0060] The ability to suppress CD38 expression is an indicator for evaluating anti-aging and skin health improvement. In this experiment, it was confirmed whether CD 38 expression was suppressed when the composition of the present invention was treated, and the results are shown in Table 1.

[0061] Specifically, fibroblasts (HS68) were seeded in 12-well plates and cultured for 24 hours. DMEM (Dulbecco's Modified Eagle Medium, Solbiopharm, Korea) was used as the culture medium, and an incubator maintained at 37°C and 5% CO2 was used. After 24 hours, the medium was removed, and the cells were washed once with PBS (Phosphate Buffered Saline). After removing the PBS, DMEM containing 25 ppm NAD+ and each of the evaluation targets according to Table 1 was added to the cells. After 24 hours, the medium was removed again, RNA was extracted (AccuPrep® Universal RNA Extraction Kit, Bioneer, Korea), and cDNA was synthesized using the RNA and cDNA synthesis kit (TOPscript™ cDNA Synthesis Kit (dN6 Mix), Enzynomics, Korea). Next, RT-qPCR (Real-time polymerase chain reaction) was performed using TOPreal™ SYBR Green RT-qPCR Kit and RT-qPCR equipment (Applied Biosystems™ StepOne™ Real-Time PCR System, Thermofisher, USA), and the expression level of CD38 was analyzed. At this time, in order to confirm the effect of the evaluation target, DMEM (Control) containing 25 ppm of NAD+ but not the evaluation target component was also performed in the same way, and the expression level of CD38 was analyzed. Then, the CD38 inhibition rate compared to the control was calculated using the following formula and is shown in Table 1.

[0062] CD38 inhibition rate (%) = [1 - (result value of evaluation target ÷ result value of control]] × 100

[0063] Evaluation target Inhibition rate [%] Comparative example 1 Quercetin 1 ppm 6.38 Comparative example 3 Glycyrrhetinic acid 5 ppm 10.64 Comparative example 5 Sitosterol 5 ppm 10.80 Example 6 Quercetin 1 ppm + Glycyrrhetinic acid 5 ppm + Sitosterol 5 ppm 64.15 Comparative example 6 Tocopheryl acetate 1 ppm 15.61 Example 8 Quercetin 1 ppm + Glycyrrhetinic acid 5 ppm + Tocopheryl acetate 1 ppm 42.53

[0064] As shown in Table 1 above, it was confirmed that CD38, which is expressed by NAD+, was significantly inhibited when quercetin, glycyrrhetinic acid, and sitosterol or tocopheryl acetate were combined with it. In particular, it was confirmed that Examples 6 and 8 of the present invention were effective in preventing and improving skin aging because the inhibition rate was remarkably high.

[0065] Experimental Example 2: Confirmation of increased intracellular NAD+

[0066] NAD+ is a factor involved in intracellular metabolism and is associated with aging. The NAD+ / NADH ratio is an indicator of how much NAD+ remains unchanged as NADH. A higher value indicates a greater anti-aging effect. In this experimental example, whether the composition of the present invention increased or decreased this indicator is shown in Table 2.

[0067] Specifically, fibroblasts (HS68) were seeded in 12-well plates and cultured for 24 hours. DMEM (Dulbecco's Modified Eagle Medium, Solbiopharm, Korea) was used as the culture medium, and an incubator maintained at 37°C and 5% CO2 was used. After 24 hours, the medium was removed, and the cells were washed once with PBS (Phosphate Buffered Saline). After removing the PBS, DMEM containing 25 ppm NAD+ and each of the evaluation targets according to Table 2 were added to the cells. After 24 hours, the medium was removed, and the cells were washed three times with PBS. Intracellular NAD+ and NADH were quantified by absorbance at 450 nm using a kit (PicoSens™ NAD / NADH Assay Kit, Biomax, Korea) according to the manufacturer's protocol, and the NAD+ / NADH ratio of each evaluation target was calculated. At this time, to confirm the effect of the evaluation target, DMEM (Control) containing 25 ppm NAD+ but not the evaluation target component was also performed in the same manner. Then, the improvement rate of NAD+ / NADH ratio compared to the control was calculated using the following formula and is shown in Table 2.

[0068] NAD+ / NADH ratio improvement rate (%)

[0069] = [(Result value of evaluation target ÷ Result value of control)-1] × 100

[0070] Evaluation target Improvement rate [%] Comparative example 1 Quercetin 1 ppm 22.54 Comparative example 2 Glycyrrhetinic acid 1 ppm 5.63 Comparative example 3 Glycyrrhetinic acid 5 ppm 36.90 Comparative example 4 Sitosterol 1 ppm 4.39 Comparative example 5 Sitosterol 5 ppm 5.04 Comparative example 7 Tocopheryl acetate 10 ppm 74.65 Example 1 Quercetin 1 ppm + Glycyrrhetinic acid 1 ppm 53.76 Example 2 Quercetin 1 ppm + Glycyrrhetinic acid 5 ppm 66.55 Example 4 Quercetin 1 ppm + Glycyrrhetinic acid 1 ppm + Sitosterol 5 ppm 72.39 Example 5 Quercetin 1 ppm + Glycyrrhetinic acid 5 ppm + Sitosterol 1 ppm60.56Example 6Quercetin 1ppm + Glycyrrhetinic acid 5 ppm + Sitosterol 5 ppm214.29Example 7Quercetin 1ppm + Glycyrrhetinic acid 1 ppm + Tocopheryl acetate 10 ppm142.42

[0071] As shown in Table 2 above, it was confirmed that the NAD+ / NADH ratio increased beyond expectations in Examples 1, 2, and 4 to 7. In particular, Examples 6 and 7 showed significantly higher improvement rates, and as a result, it was confirmed that the use of the skin topical agent according to the present invention can effectively inhibit skin aging by increasing the concentration of nicotinamide adenine dinucleotide in skin cells.

[0072] Experimental Example 3: Confirmation of the cell aging inhibition effect

[0073] The degree of cell aging was increased by treating fibroblasts with oxidative stress (H2O2), and then the composition of the present invention was treated to confirm whether the degree of aging was improved, as shown in Table 3 and Figure 1.

[0074] Specifically, fibroblasts (HS68) were seeded in 96-well plates and cultured for 24 hours. DMEM (Dulbecco's Modified Eagle Medium, Solbiopharm, Korea) was used as the culture medium, and an incubator maintained at 37°C and 5% CO2 was used. After 24 hours, the medium was removed, and the cells were washed once with PBS (Phosphate Buffered Saline). After removing the PBS, DMEM containing 25 ppm NAD+ and each evaluation target according to Table 3 was added to the cells. After 4 hours, 300 μM (final concentration) of hydrogen peroxide (H2P) was treated to the cells, and after 2 hours of incubation, the medium was removed, and the cells were washed twice with PBS. Then, DMEM containing each evaluation target according to Table 3 was added to the cells, and the cells were cultured for 24 hours. After 24 hours, the medium was removed, and the cells were washed once with PBS. Afterwards, the kit for measuring cellular senescence (Cellular Senescence Plate Assay Kit - SPiDER-β Gal, Dojindo, Japan) was used according to the manufacturer's protocol, and the degree of cellular senescence was quantified by measuring the fluorescence intensity under the conditions of Ex: 535 nm; Em: 580 nm. At this time, in order to confirm the effect of the evaluation target, the experimental group in which only DMEM was added to the cells without the evaluation target and the step of adding hydrogen peroxide (negative control group, N.Control), the experimental group in which only DMEM was added to the cells without the evaluation target and the step of adding hydrogen peroxide (reference value, Reference), and DMEM containing 25 ppm of NAD+ but not the evaluation target component (Control) were performed in the same way. Then, the senescence improvement rate compared to the reference value was calculated using the following formula.

[0075] Cell aging improvement rate (%) =

[0076] [1 - (Result value of evaluation target ÷ Result value of reference value]] × 100

[0077] In an experiment to visualize the degree of senescence, designed to intensify green fluorescence as senescence deepens, a staining kit for SA-bgal, an indicator of cellular senescence (Cellular Senescence Detection Kit - SPiDER-ßGal, Dojindo, Japan) was used. Staining was performed according to the manufacturer's protocol and observation was performed through a fluorescence microscope (EVOS™ M7000, Thermofisher, USA), and the results of treatment with the composition of the present invention are shown in Fig. 1.

[0078] Evaluation target Improvement rate [%] Comparative example 1 Quercetin 1 ppm 12.43 Comparative example 2 Glycyrrhetinic acid 1 ppm 5.41 Comparative example 3 Glycyrrhetinic acid 5 ppm 12.43 Comparative example 4 Sitosterol 1 ppm 2.16 Comparative example 5 Sitosterol 5 ppm 5.41 Comparative example 6 Tocopheryl acetate 1 ppm 8.11 Comparative example 7 Tocopheryl acetate 10 ppm 9.19 Example 1 Quercetin 1 ppm + Glycyrrhetinic acid 1 ppm 18.92 Example 3 Quercetin 1 ppm + Glycyrrhetinic acid 1 ppm + Sitosterol 1 ppm 29.19 Example 4 Quercetin 1 ppm + Glycyrrhetinic acid 1 ppm + Sitosterol 5 ppm37.84Example 8Quercetin 1 ppm + Glycyrrhetinic acid 5 ppm + Tocopheryl acetate 1 ppm32.43Example 9Quercetin 1 ppm + Glycyrrhetinic acid 5 ppm + Tocopheryl acetate 10 ppm34.05

[0079] As shown in Table 3 and Fig. 1, when oxidative stress (hydrogen peroxide treatment) was increased in fibroblasts and treatment with Examples 1, 3, 4, 8, and 9 of the present invention was performed to determine whether cell aging was improved, a synergistic effect exceeding expectations was confirmed, and in particular, when Example 4 was applied, it was confirmed that green fluorescence induced by aging almost disappeared, as shown in Fig. 1. Therefore, it was confirmed that the cosmetic composition of the present invention is effective in inhibiting cell aging.

[0080] Experimental Example 4: Confirmation of MMP-1 expression inhibition by UV rays

[0081] It was confirmed whether MMP-1 expression was inhibited when the composition of the present invention was treated after increasing cellular MMP-1 expression following UV treatment.

[0082] Specifically, fibroblasts (HS68) were seeded in 12-well plates and cultured for 24 hours. DMEM (Dulbecco's Modified Eagle Medium, Solbiopharm, Korea) was used as the culture medium, and an incubator maintained at 37°C and 5% CO2 was used. After 24 hours, the medium was removed and the cells were washed once with PBS (Phosphate Buffered Saline). After removing the PBS, DMEM containing 25 ppm NAD+ and each of the evaluation targets according to Table 4 were added to the cells. After 4 hours of culture, the medium was removed, PBS was administered to the cells, and then UV irradiation was performed. UV irradiation was performed using a BIO-SUN UV irradiator (Vilber Lourmat, France) at 312 nm narrow-band UVB at 35 mJ / cm2 and 365 nm narrow-band UVA at 15 mJ / cm2. After this, PBS was removed, and each DMEM containing the evaluation target according to Table 4 was added to the cells again, and cultured for 48 hours. After 48 hours, the medium supernatant was harvested, and the MMP-1 expression amount was quantified through ELISA (Enzyme-linked immunosorbent assay). For this, Human Total MMP-1 DuoSet ELISA (R&D systems, USA) was used, and the experiment was conducted according to the manufacturer's guide. Protein quantification was performed using BCA assay (Pierce™ BCA Protein Assay Kits, Thermofisher, USA). At this time, in order to confirm the effect of the evaluation target, an experimental group (negative control group, N.Control) in which only DMEM without the evaluation target was added to the cells and the UV irradiation step was not performed, and DMEM containing 25 ppm of NAD+ but not the evaluation target component (Control) was also performed in the same way.And the MMP-1 expression inhibition ability compared to the control was calculated by the following formula.

[0083] MMP-1 expression inhibition ability (%) =

[0084] [1 - (Result value of evaluation target ÷ Result value of control]] × 100

[0085] Evaluation target Improvement rate [%] Comparative example 1 Quercetin 1 ppm 5.58 Comparative example 2 Glycyrrhetinic acid 1 ppm 14.55 Example 1 Quercetin 1 ppm + Glycyrrhetinic acid 1 ppm 30.18

[0086] As shown in Table 4, when fibroblasts were treated with UV rays to increase MMP-1 expression and Example 1 of the present invention was used to confirm the prevention and improvement rate of cell aging, it was confirmed that Example 1 had an increased ability to suppress MMP-1 expression. Therefore, it was confirmed that the cosmetic composition of the present invention is effective in preventing skin aging and maintaining elasticity.

[0087] Experimental Example 5: Confirmation of the effect of extending cell replication lifespan

[0088] It was confirmed whether the composition according to the present invention has the effect of extending cell division ability and replicative lifespan.

[0089] Specifically, fibroblasts (HS68) were seeded in a 96-well plate at 10 cells and cultured for 24 hours. DMEM (Dulbecco's Modified Eagle Medium, Solbiopharm, Korea) was used as the culture medium, and an incubator maintained at 37℃ and 5% CO2 was used. After 24 hours, the medium was removed and washed once with PBS (Phosphate Buffered Saline). After removing the PBS, DMEM containing 25 ppm of NAD+ and each of the evaluation targets according to Table 5 were added to the cells. Thereafter, the medium was replaced every 7 days, and the cell population and proliferation were measured on days 0, 21, and 28. At this time, in order to confirm the effect of the evaluation target, the negative control group (N.Control) treated only with DMEM and DMEM (Control) containing 25 ppm of NAD+ but not the evaluation target component were performed and measured in the same way. Measurements were performed using the CCK-8 assay (CK04, Dojindo, Japan), and the results are shown in Fig. 2. At this time, the relative cell population was proportional to the cell number, and the effects of extending cell division ability and replicative lifespan based on the measured values ​​were calculated according to the following formula and shown in Table 5.

[0090] Effect (%) = [1 - (Result of evaluation target ÷ Result of N.Control]] × 100

[0091]

[0092] Evaluation target Improvement rate (%, 21st day) Improvement rate (%, 28th day) Control - 13.3 26.6 Example 6 Quercetin 1 ppm + Glycyrrhetinic acid 5 ppm + Sitosterol 5 ppm 34.1 8 2.3 Example 7 Quercetin 1 ppm + Glycyrrhetinic acid 1 ppm + Tocopheryl acetate 10 ppm 30.0 7 1.1

[0093] As shown in Table 5 and Figure 2, when triple-combined as in Examples 6 and 7, the effects of extending cell division ability and replicative lifespan were confirmed. Therefore, it was confirmed that the cosmetic composition of the present invention is effective in inhibiting cell aging, improving skin texture, regenerating skin, and strengthening the skin barrier.

[0094] In summary of the embodiments and experimental examples of the present invention, it was confirmed that the cosmetic composition of the present invention provides an effect of improving aging of damaged skin and an effect of improving skin wrinkles and elasticity by increasing the concentration of nicotinamide adenine dinucleotide in skin cells.

[0095]

[0096] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. A cosmetic composition comprising nicotinamide adenine dinucleotide; glycyrrhetinic acid; and quercetin.

2. In paragraph 1, A cosmetic composition further comprising sitosterol or tocopheryl acetate.

3. In paragraph 1, A cosmetic composition wherein the nicotinamide adenine dinucleotide and glycyrrhetinic acid are included in a weight ratio of 1:0.0004 to 1:0.

4.

4. In paragraph 1, A cosmetic composition wherein the nicotinamide adenine dinucleotide and quercetin are included in a weight ratio of 1:0.0004 to 1:0.

4.

5. In paragraph 2, A cosmetic composition wherein the nicotinamide adenine dinucleotide and sitosterol are included in a weight ratio of 1:0.0004 to 1:0.

4.

6. In paragraph 2, A cosmetic composition wherein the nicotinamide adenine dinucleotide and tocopheryl acetate are included in a weight ratio of 1:0.0004 to 1:

1.

7. In paragraph 1, The above composition is a cosmetic composition for inhibiting skin aging.

8. In paragraph 1, The above composition is a cosmetic composition for inhibiting photoaging.

9. A method for inhibiting skin aging, comprising the step of applying a cosmetic composition comprising nicotinamide adenine dinucleotide; glycyrrhetinic acid; and quercetin to the skin of a subject.

10. A method for inhibiting photoaging, comprising the step of applying a cosmetic composition comprising nicotinamide adenine dinucleotide; glycyrrhetinic acid; and quercetin to the skin of a subject.

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