Composition comprising TES trioleate and oleamide serinol as active ingredients for alleviating and preventing cellular senescence

Abstract

The present invention relates to a composition comprising TES trioleate and oleamide serinol as active ingredients for alleviating and preventing cellular senescence. According to the present invention, the composition comprising TES trioleate and oleamide serinol exhibits cytoprotective effects on skin cells damaged by UV and inhibitory effects on generation of reactive oxygen species. According to the present invention, the composition comprising TES trioleate and oleamide serinol exhibits the effects of protecting skin cells damaged by UV, suppressing generation of senescence-associated marker proteins (SA-β-gal), and inhibiting formation of a senescence-associated secretory phenotype (SASP).

Description

A composition for alleviating and preventing cellular aging comprising TES TRIOLEATE and OLEAMIDE SERINOL as active ingredients

[0001] The present invention relates to a composition for alleviating and preventing cellular aging comprising TES Trioleate and Oleamide Serinol as active ingredients.

[0002] Population aging is currently a global megatrend. As aging progresses, the body's ability to recover from stress gradually declines. Since aging also causes serious economic problems, interest in aging research is surging worldwide.

[0003] Cellular senescence, one of the major mechanisms of aging, is an irreversible cell cycle arrest resulting from stress or stimulation; that is, a state where cell division has stopped but the cells have not died. Cells in this state are called senescent cells or zombie cells. As aging occurs, the immune system loses the ability to eliminate zombie cells, and as zombie cells accumulate, they release 'inflammatory chemicals with toxic effects' to nearby healthy cells.

[0004] Senotherapeutics is a new class of natural or synthetic compounds that can help the immune system alleviate the effects of the presence of senescent cells in aging-related diseases, and research is being conducted on (1) senolytics that selectively remove senescent cells; and (2) senomorphic compounds that enhance cell function or inhibit pathways associated with the expression of aging-related secretory phenotype (SASP) components.

[0005] As prior art related to senomorphic, KR10-2687771 discloses a combination composition for senomorphic that restores senescent cells, whose cell cycle has been arrested due to aging, to the level of young cells when treated in combination, and KR10-2618160 discloses that rambutan seed extract is effective in inhibiting symptoms related to cell aging. Additionally, KR10-2022-0161172 discloses a composition for preventing skin aging comprising arginine and HEPES{4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid}.

[0006] It has been studied that testriooleate has a skin protective effect (KR10-1899345) by inhibiting the activity of PLA2 (Phospholipase A2) in red blood cells or keratinocytes in the skin, thereby inhibiting the generation of cytotoxic metabolites caused by the oxidation of UVA or reactive oxygen species, but no cenomorphic effect has been disclosed regarding the inhibition of aging-related symptoms in aged cells.

[0007] [Prior Art Literature]

[0008] [Patent Literature]

[0009] KR10-2687771

[0010] KR10-2618160

[0011] KR10-2022-0161172

[0012] The present invention is intended to provide a composition capable of alleviating or preventing cellular aging.

[0013] To achieve the above objective, the present invention provides a composition for alleviating and preventing cellular aging comprising testioleate and oleamide sulinol as active ingredients.

[0014] According to one embodiment of the present invention, the weight ratio of the testriooleate and oleamide sulinol may be 1:1 to 1:3.

[0015] According to one embodiment of the present invention, the composition may further include one or more of echinacea extract or fermented yuzu extract.

[0016] According to one embodiment of the present invention, the content of the echinacea extract may be 1:1 to 1:3 in weight relative to the testioleate.

[0017] According to one embodiment of the present invention, the content of the fermented yuzu extract may be 1:1 to 1:5 relative to the weight of the testioleate.

[0018] According to one embodiment of the present invention, the composition for alleviating and preventing cell aging may be a cenomorph that restores the function or morphology of aging cells to the function or morphology of normal cells.

[0019] According to one embodiment of the present invention, the composition for alleviating and preventing cell aging may be a pharmaceutical composition.

[0020] According to one embodiment of the present invention, the composition for alleviating and preventing cell aging may be a quasi-drug composition.

[0021] According to one embodiment of the present invention, the composition for alleviating and preventing cell aging may be a cosmetic composition.

[0022] According to the present invention, a composition comprising testioleate and oleamide sulinol has a cytoprotective effect on skin cells damaged by UV and an effect of inhibiting the production of reactive oxygen species.

[0023] According to the present invention, a composition comprising testriooleate and oleamide sulinol has the effect of inhibiting the production of senescent cell marker protein (SA-β-gal) and the production of senescence-related secretory phenotype (SASP) in cells, such as epidermal cells and dermal cells, that are damaged by UV radiation.

[0024] Meanwhile, the scope of the present invention is not limited by the effects described above.

[0025] The present invention will be described in detail as follows.

[0026] Meanwhile, each description and embodiment disclosed in this application may also be applied to other descriptions and embodiments. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application should not be considered limited by the specific descriptions provided below.

[0027]

[0028] The present invention relates to a composition for alleviating and preventing cellular aging, comprising TES Trioleate and Oleamide Serinol as active ingredients.

[0029] The above testriooleate is a substance that has a skin-protective effect by inhibiting PLA2 (Phospholipase A2) activity, and it was confirmed that when combined with specific ingredients, it can improve aging-related symptoms observed in senescent cells by reducing the increased beta-galactosidase protein in senescent cells and inhibiting SASP, an aging-related secretory phenotype.

[0030] Generally, aged skin cells exhibit slow cell division and an increase in senescence-associated β-galactosidase (SA-β-gal) in culture and tissue samples, and when examined with the naked eye, the cells become larger and exhibit a flattened cell shape.

[0031] In addition, senescent cells secrete a senescence-associated secretory phenotype (SASP) into the surrounding environment, which includes multiple soluble factors related to growth factors and IL-1, IL-6, IL-8, proteases, and matrix metalloproteinases (MMPs) as indicators of cellular senescence; ultimately, SASP disrupts the local tissue environment and induces inflammation in various tissues and organs, thereby causing skin cell senescence.

[0032] It is preferable that the testioleate be included in an amount of 0.01 to 10 weight% relative to the total weight of the composition, and more preferable that it be included in an amount of 0.01 to 5 weight%. If the content of the testioleate is less than 0.01 weight%, there is a concern that the inhibitory effect on SASP may not be sufficient, and if it exceeds 10 weight%, it may cause formulation problems.

[0033]

[0034] The above-mentioned oleamide serinol is a ceramide-like compound in which 2-amino-1,3-propanediol and oleic acid are chemically bonded; it promotes bioactivity to enhance skin elasticity and also exerts antioxidant and anti-inflammatory effects.

[0035] In one embodiment of the present invention, the oleamide sulinol may be N-(1,3-dihydroxypropane-2-nyl)oleamide.

[0036] It is preferable that the oleamide serinol comprises 0.01 to 10 weight% based on the total weight of the composition, and more preferable that it comprises 0.01 to 5 weight%. If the content of the oleamide serinol is less than 0.01 weight%, there is a concern that it may not be sufficient to exhibit the effect of alleviating and preventing cell aging, and if it exceeds 10 weight%, it may cause formulation problems.

[0037] The weight ratio of the above testriooleate and oleamide sulinol is preferably 1:1 to 1:3. If the weight ratio of testriooleate and oleamide sulinol is less than 1:1 or exceeds 1:3, the anti-aging and preventive effects may be insufficient.

[0038]

[0039] Preferably, the composition may further include one or more of echinacea extract or fermented yuzu extract to enhance the cellular aging alleviation and prevention effects intended by the present invention.

[0040] Echinacea extract may have an improving effect on at least one of skin damage caused by pollutants, skin contamination, skin cell death, inhibition of skin cell proliferation, and deterioration of skin condition.

[0041] If echinacea extract is further included, the content of echinacea extract is preferably 1:1 to 1:3 in weight relative to testioleate.

[0042] Fermented yuzu extract can provide skin moisturizing, skin barrier strengthening, antioxidant, and wound healing effects for damaged skin cells.

[0043] If fermented yuzu extract is further included, the content of fermented yuzu extract is preferably 1:1 to 1:5 relative to the weight of testioleate.

[0044] The above composition for alleviating and preventing cellular aging may be a cenomorph that inhibits SASP to restore the function or morphology of aging cells to the function or morphology of normal cells, and the cellular aging may be skin cell aging caused by UV.

[0045] In addition, the above composition for alleviating and preventing cell aging may utilize a transdermal drug delivery system (TDDS) that delivers drugs through the skin, such as liposomes, lysosomes, exosomes, vesicles, and microneedles, to improve skin permeability.

[0046] In addition, the above composition for alleviating and preventing cell aging may be a pharmaceutical composition.

[0047] The above pharmaceutical composition may be a pharmaceutical composition for skin protection against UV.

[0048] The above skin protection may be achieved by preventing skin cell damage, and for example, the skin cells may be keratinocytes (HaCaT cells) or fibroblasts (NHDF cells) within the skin.

[0049] Specific formulations of the above pharmaceutical composition may include, for example, formulations such as hydrogels, emulsion creams, liposome-containing hydrogels, ointments, liquids, or suspensions.

[0050] To summarize the above, TES Trioleate can not only prevent and protect against skin cell damage by inhibiting the activity of PLA2 caused by UVA and reactive oxygen species, but also, when combined with specific ingredients, can reduce senescent cells and activate normal cells through the removal of senescent cells or the inhibition of SASP expression, thereby alleviating, preventing, and preserving skin cell aging.

[0051] In addition, the above composition for alleviating and preventing cell aging may be a quasi-drug composition.

[0052] In the present invention, 'quasi-drug' may include, in one embodiment, an oral formulation, but is not limited thereto, and the formulation method, dosage, method of use, components, etc. of the quasi-drug may be appropriately selected from ordinary techniques known in the technical field.

[0053] The quasi-drug composition of the present invention may further include, in addition to the above components, a pharmaceutically acceptable carrier, excipient, or diluent as needed. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effects of the present invention, and may include, for example, fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc.

[0054]

[0055] In addition, the above composition for alleviating and preventing cell aging may be a cosmetic composition.

[0056] The above cosmetic composition may be a cosmetic composition for preventing photoaging. The above photoaging refers to skin aging caused by ultraviolet rays in extrinsic aging. The above photoaging prevention may be due to the prevention of or protection of skin cell damage, and for example, the skin cells may be keratinocytes (HaCaT cells) or fibroblasts (NHDF cells) within the skin.

[0057] The above cosmetic composition may further include ingredients commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, or fragrances.

[0058] The above cosmetic composition can be manufactured in various forms, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, oil, powder foundation, or emulsion foundation.

[0059] Specific formulations of the above cosmetic composition may include, for example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, essence, nourishing essence, pack, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, lipstick, makeup base, foundation, pressed powder, loose powder, or eyeshadow.

[0060] It is obvious that the effect is the same even when the composition of the present invention is applied to pharmaceutical compositions, quasi-drug compositions, and cosmetic compositions. It can be used in compositions for protecting the skin from ultraviolet rays, for example, compositions for protecting the skin by preventing photoaging, or it can be used in compositions for tanning or sunscreens to exhibit a skin protection effect.

[0061] The present invention will be explained in more detail below through the following examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0062]

[0063] <Preparation Example>

[0064] Preparation Example 1. Preparation of TES Trioleate

[0065] 7.0 kg of 2-[tris(hydroxymethyl)-methylamino]-1-ethanesulfonic acid, 38.3 kg of dimethylacetamide, and 37.0 kg of oleoyl chloride were added to a stirrer, heated to 65±5℃, and stirred for 3 hours. Once the reaction was complete, 70.0 kg of water and 62.8 kg of ethyl acetate were added for extraction; the organic layer was then washed with saturated brine, dried with magnesium sulfate, filtered, and concentrated. The concentrate was recrystallized under acetone to obtain 24 kg of TES trioleate (yield: 77.8%).

[0066] Preparation Example 2. Preparation of Oleamide Sullinol (N-(1,3-dihydroxypropane-2-nyl)oleamide)

[0067] 25.51 g of serinol and 47 ml of triethylamine were placed in a reaction flask, followed by 100 ml of ethanol and 250 ml of dichloromethane, and the temperature was lowered to a low temperature (approx. 5°C). A solution of 104 ml of oleyl chloride and 100 ml of dichloromethane was then added dropwise to the reaction flask. The reaction solution was stirred at room temperature for 6 hours, after which the solution was concentrated under reduced pressure. The concentrated liquid was dissolved in 500 ml of dichloromethane and washed with 0.5 N aqueous hydrochloric acid solution, saturated aqueous sodium bicarbonate, and water. The organic solution layer was dried with anhydrous magnesium sulfate and filtered; the organic solvent was then concentrated under reduced pressure and recrystallized to obtain 83 g (83%) of a pale yellow solid.

[0068] Preparation Example 3. Preparation of Echinacea Extract

[0069] For the hot water extract of Echinacea (stem, leaf, flower), 40 g of fine powder sample was prepared by grinding Echinacea (stem, leaf, flower) washed with distilled water using a blender, adding 4 L of distilled water, and performing hot water extraction while heating at 100°C for 3 hours. After performing the extraction, the Echinacea extract was prepared by filtering through a 0.45 μm filter paper.

[0070] Preparation Example 4. Preparation of Fermented Yuzu Extract

[0071] The entire pulp of the yuzu was dried and ground, and purified water was added in an amount 10 to 30 times the volume of the sample. Hot water extraction was performed at 60 to 100°C for 1 to 5 hours. After performing the extraction, the mixture was filtered through a 0.45 μm filter paper to prepare a yuzu extract. Lactobacillus rhamnosus was added to the yuzu extract in an amount of 0.05 to 3%, and a fermentation process was carried out at 32 to 37°C for 3 to 5 days. The fermented liquid was then filtered to prepare a fermented yuzu extract.

[0072] <Example>

[0073] Examples 1 to 4

[0074] A composition was prepared by mixing the materials prepared in the above Preparation Examples 1 to 4 according to the following composition ratios.

[0075] Comparative Example 1

[0076] A composition containing the TES Trioleate prepared in Preparation Example 1 above was prepared.

[0077]

[0078] (Weight%) TES Trioleate Oleamide Sullinol Echinacea Extract Fermented Yuzu Extract Comparative Example 11 Example 112 Example 2121 Example 3122 Example 41223

[0079] <Experimental Example>

[0080] Experimental Example 1. Cytoprotective effect on skin cells damaged by UV radiation

[0081] The composition obtained in the above example was evaluated to determine whether it could protect skin cells damaged by UV-B stimulation. HaCaT keratinocytes were used as epidermal keratinocytes, and NHDF (normal human dermal fibroblast) was used as dermal cells.

[0082] HaCaT cells, a human keratinocyte cell line, were obtained from Dr. Fusenig (German cancer research center, DKFZ) and cultured. The cell culture medium DMEM (Dulbecco's modified Eagle's medium) was prepared by mixing 10% fetal bovine serum (FBS, PAA, Austria) and 1% penicillin-streptomycin (PAA, Austria).

[0083] 5 × 10⁶ HaCaT and NHDF cells were each placed in a 24-well plate. 4 Cells were seeded per well and cultured for 24 hours at 37°C under 5% CO2 conditions. After removing the cultured medium and washing with PBS, 300 μL of PBS was injected into each well, followed by irradiation with 25 mJ of UVB. After removing the PBS, the compositions of Examples 1 to 4 were each treated at a concentration of 0.10%, and the cells were cultured for 48 hours. The control group was treated with Quercetin diluted to 1 μM. After the culture was complete, EZ-Cytox was added to each well to make up 10% of the medium, and the cells were cultured for an additional 1 hour. The cultured medium was transferred to a 96-well plate, and SpectraMax ® Absorbance was measured at 450 nm using an iD5 microplate reader (Molecular Devices, USA), and the cytoprotective effect of the compositions of Examples 1 to 4 was evaluated using the following Equation 1, and the results are shown in Table 2 below.

[0084] [Mathematical Formula 1]

[0085] Cell viability (%) = (Average ODs / Average OD) UVB ) X 100

[0086] ODs: Absorbance measurements of the sample

[0087] OD UVB: Measured absorbance of the UVB-treated control group

[0088]

[0089] Treatment Concentration (%) HaCaT (%) NHDF (%) Control - 136.35 121.07 UVB Treatment Control Group - 100.00 100.00 Comparative Example 10.10 11.12 95.97 Example 10.10 116.55 91.98 Example 20.10 118.07 93.18 Example 30.10 117.34 96.44 Example 40.10 121.09 102.67 Quercetin (μM) 1.00 110.39 113.29

[0090]

[0091] As shown in Table 2 above, when the cytoprotective effect of the compositions of Examples 1 to 4 was evaluated on UV-stimulated epidermal keratinocytes and dermal fibroblasts, it was found that while the cytoprotective effect in the dermal layer was negligible, the cytoprotective effect in the epidermal layer increased compared to the UV-treated group. The compositions of Examples 1 to 4 containing TES trioleate and oleamide sulinol showed superior cytoprotective effects compared to the composition of Comparative Example 1, and in particular, the compositions of Examples 2 to 4 showed even better efficacy.

[0092] Experimental Example 2. Inhibitory effect on the production of reactive oxygen species in skin cells damaged by UV radiation

[0093] The composition obtained in the above example was measured to determine whether it could suppress reactive oxygen species (ROS) that increase in skin cells damaged by UV-B stimulation. HaCaT keratinocytes were used as epidermal keratinocytes, and NHDF (normal human dermal fibroblast) was used as dermal cells.

[0094] HaCaT and NHDF cells were placed in 5×10⁶ wells of a 24-well plate, respectively. 4 cells / well or 2×10 4Cells were seeded into cells / well and cultured at 37°C under 5% CO2 conditions for 24 hours. Subsequently, the compositions of Examples 1 to 4 were each treated at a concentration of 0.01% and cultured for 24 hours. After removing the culture medium containing the samples and washing with PBS, 300 μL / well of PBS was injected to irradiate HaCaT cells with 150 mJ of UVB and NHDF cells with 170 mJ of UVB. Afterward, the medium was changed and cultured for an additional 3 hours. The control group was treated with 20 μM of Luteolin.

[0095] The amount of intracellular reactive oxygen species (ROS) produced is Oxiselect TM After measuring using the Intracellular ROS assay Kit (CELL BIOLABS, INC., USA), the intracellular reactive oxygen species (ROS) inhibitory effect according to the compositions of Examples 1 to 4 was evaluated through the following mathematical formula 2, and the results are shown in Table 3 below.

[0096] [Mathematical Formula 2]

[0097] Reactive oxygen species inhibition rate (%) = (ROSs / ROS UVB ) X 100

[0098] ROSs: Absorbance measurements of the corrected sample

[0099] ROS UVB : Absorbance measurement of the corrected UV-treated control

[0100]

[0101] Treatment Concentration (%) HaCaT (%) NHDF (%) Control - 68.59 67.64 UVB Treatment Control Group - 100.00 100.00 Example 10.01 81.5 180.44 Example 20.01 76.34 75.71 Example 30.01 79.5 176.92 Example 40.01 72.78 62.13 Luteolin (μM) 20.06 0.47 60.27

[0102]

[0103] As confirmed in Table 3 above, the inhibition of reactive oxygen species induced by UV stimulation in UV-stimulated keratinocytes (HaCaT) and fibroblasts (NHDF) was evaluated for the compositions of the present invention, and the amount of reactive oxygen species produced, which had increased due to UV, was found to be reduced. The compositions of Examples 1 to 4 containing TES trioleate and N-(1,3-dihydroxypropane-2-nyl)oleamide showed excellent reactive oxygen species inhibition effects, and in particular, the compositions of Examples 2 to 4 showed even better efficacy.

[0104] Experimental Example 3. Effects of inhibiting the production of senescent cell marker protein (SA-β-gal) and on the senescence-associated secretory phenotype (SASP) in UV-damaged epidermal cells

[0105] The composition obtained in the above example was measured to determine whether it had an effect on the production of senescence-associated β-galactosidase (SA-β-gal) and senescence-associated secretory phenotype (SASP), which are increased in skin cells damaged by UV-B stimulation. HaCaT keratinocytes were used as epidermal keratinocytes.

[0106] 5 × 10⁶ HaCaT cells in a 24-well plate 4Cells were seeded into wells and cultured at 37°C under 5% CO2 conditions for 24 hours. Subsequently, the compositions of Examples 1 to 4 were each treated at a concentration of 0.05% and cultured for 24 hours. The culture medium containing the samples was removed, washed with PBS, and 300 μL / well of PBS was injected to irradiate with 25 mJ of UVB. The medium was then replaced, and the cells were cultured for an additional 24 hours. The control group was treated with 1 μM of retinoic acid. After the culture was completed, the culture medium was taken, and the amount of Interleukin-6 (IL-6) produced in the medium was quantified using the Human IL-6 Quantikine ELISA Kit (R&D Systems, USA). The cultured cells were stained for the senescence marker (SA-β-gal) using the Cellular Senescence Assay Kit (SA-β-gal Staining) (CELL BIOLABS, INC., USA), and the senescence marker (SA-β-gal) was measured using an inverted microscope. The measured images were used to determine the degree of cellular aging through the ratio of stained cells to total cells, and the results were quantified and shown in Table 4 below.

[0107] [Mathematical Formula 3]

[0108] Inhibition rate of aging cell marker generation (%) = (DC s / AC s ) / ( DC UVB / AC UVB )×100

[0109] * DC S : Number of stained senescent cells in the example

[0110] * AC s : Total number of cells treated in the examples

[0111] * DC UVB : Number of senescent cells stained in the UVB-treated control group

[0112] * AC UVB : Total number of cells in the UVB-treated control group

[0113]

[0114] Treatment Concentration (%) SASP (IL-6) Production Inhibition Rate (%) Aging Cell Marker (SA-β-gal) Production Inhibition Rate (%) Control - 13.6675.06 UVB Treatment Control Group - 100.00100.00 Comparative Example 10.0590.9795.12 Example 10.0579.4792.48 Example 20.0572.1288.30 Example 30.0570.4778.03 Example 40.0564.4161.38 Retinoic Acid (μM) 1.0061.6460.15

[0115]

[0116] As confirmed in Table 4 above, the inhibition of interleukin-6 (IL-6) production among the aging-related secretory phenotypes (SASP) in epidermal cells stimulated by UV with the composition of the present invention was evaluated, and the amount of IL-6 production, which was increased by UV, was found to be reduced. In addition, the inhibition of the production of the aging cell marker (SA-β-gal) induced by UV stimulation in epidermal cells stimulated by UV with the composition of the present invention was evaluated, and the aging cell marker (SA-β-gal), which was increased by UV, was found to be inhibited. The compositions of Examples 1 to 4 containing TES Trioleate and N-(1,3-dihydroxypropane-2-nyl)oleamide showed excellent inhibitory effects on IL-6 and SA-β-gal production, and in particular, the compositions of Examples 2 to 4 showed even better inhibitory efficacy on IL-6 and SA-β-gal production.

[0117] Experimental Example 4. Effects of inhibiting the production of senescent cell marker protein (SA-β-gal) in UV-damaged dermal cells and on the aging-associated secretory phenotype (SASP).

[0118] The composition obtained in the above example was measured to determine whether it had an effect on the production of senescence-associated β-galactosidase (SA-β-gal) and senescence-associated secretory phenotype (SASP), which are increased in skin cells damaged by UV-B stimulation. Human dermal fibroblasts (HDF) were used as dermal cells.

[0119] 1.5 × 10⁶ HDF cells in a 48-well plate 4 Cells were seeded into wells and cultured at 37°C under 5% CO2 conditions for 24 hours. After removing the culture medium and washing with PBS, 300 μl / well of PBS was injected, and UVB 55 mJ was irradiated. After removing the PBS, the compositions of Examples 1 to 4 were each treated at a concentration of 0.01% and cultured for 48 hours. The control group was treated with Epigallo Catechine Gallate (EGCG) diluted to 1 μg / ml. Subsequently, the supernatants from Examples 1 to 4 were collected and quantified using an MMP-1 Human ELISA kit (Abcam Limited). The cultured cells were stained for the senescence marker (SA-β-gal) using a Cellular Senescence Assay Kit (SA-β-gal Staining) (CELL BIOLABS, INC., USA), and the senescence marker (SA-β-gal) was measured using an inverted microscope. The measured images were quantified after confirming the degree of cell aging through the ratio of stained cells to total cells, and the results of measuring the inhibition of MMP-1 production according to the compositions of Examples 1 to 4 are shown in Table 5 below.

[0120] [Mathematical Formula 4]

[0121] Inhibition rate of aging cell marker generation (%) = (DC s / AC s ) / ( DCUVB / AC UVB )×100

[0122] * DC S : Number of stained senescent cells in the example

[0123] * AC s : Total number of cells treated in the examples

[0124] * DC UVB : Number of senescent cells stained in the UVB-treated control group

[0125] * AC UVB : Total number of cells in the UVB-treated control group

[0126]

[0127] Treatment Concentration (%) SASP (MMP-1) Production Inhibition Rate (%) Aging Cell Marker (SA-β-gal) Production Inhibition Rate (%) Control - 6 2.71 38.71 UVB Treatment Control Group - 100.00 100.00 Comparative Example 10.01 95.2 79 8.84 Example 10.01 75.4 48 8.76 Example 20.01 69.4 97 7.64 Example 30.01 57.4 46 7.19 Example 40.01 43.09 41.31 EGCG (μg / ml) 1.02 5.40 - Retinoic acid (μM) 1.0-17.69

[0128]

[0129] As confirmed in Table 5 above, the inhibition of matrix metalloproteinase-1 (MMP-1) production among the UV-induced aging-related secretory phenotypes (SASP) in UV-stimulated epidermal cells with the composition of the present invention was evaluated, and the amount of MMP-1 production, which had been increased by UV, was found to be reduced. In addition, the inhibition of the production of the aging cell marker (SA-β-gal), which had been induced by UV stimulation, was evaluated in UV-stimulated dermal cells with the composition of the present invention, and the aging cell marker (SA-β-gal), which had been increased by UV stimulation, was found to be inhibited. The compositions of Examples 1 to 4 containing TES Trioleate and N-(1,3-dihydroxypropane-2-nyl)oleamide exhibited excellent inhibitory effects on MMP-1 and SA-β-gal production, and in particular, the compositions of Examples 2 to 4 exhibited even better inhibitory efficacy on MMP-1 and SA-β-gal production.

[0130]

[0131] Jejeye

[0132] Formulation Example 1: Preparation of a cream comprising testioleate (TES Trioleate) and N-(1,3-dihydroxypropane-2-nyl)oleamide

[0133] A cream for protecting skin cells from ultraviolet rays or for skin protection was prepared by mixing the following ingredients according to a conventional cream manufacturing method.

[0134] Content of Formulated Ingredients (Weight%) Purified Water UP to 100 Example 10.5 Beta-1,3-Glucan 5 Polysorbate 80 1.5 Squalane 5 Glycerin 5 Butylene Glycol 3 Propylene Glycol 3 Cetearyl Oliveate / Sorbitan Oliveate 1 Appropriate amount of preservatives, colorants, and fragrances

[0135]

[0136] Formulation Example 2: Preparation of a cosmetic lotion containing testioleate (TES Trioleate) and N-(1,3-dihydroxypropane-2-nyl)oleamide

[0137] After mixing the following ingredients, a cosmetic lotion for skin cell protection or skin protection was prepared according to a conventional manufacturing method.

[0138] Content of Formulated Ingredients (Weight%) Purified Water UP to 100 Example 30.1 Butylene Glycol 2.0 Propylene Glycol 2.0 Acrylate / C10-C30 Alkyl Acrylate Crosspolymer 0.1 Polysorbate 800.4 Arginine Acid 0.1 Xanthan Gum 0.1 Hyaluronic Acid 1.0 Appropriate Amounts of Preservatives, Colorants, and Fragrances

[0139]

[0140] Formulation Example 3: Preparation of a topical skin preparation containing testioleate (TES Trioleate) and N-(1,3-dihydroxypropane-2-nyl)oleamide

[0141] After mixing the following ingredients, an ointment for protecting skin cells or skin was prepared according to a conventional manufacturing method.

[0142] Content of blended ingredients (weight%) Purified water UP to 100 Example 41 Diethanolamine 1.5 Polyvinylpropylidone 5 Propylene glycol 30 Diethanolamine 1.5

[0143]

[0144] From the foregoing description, those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in other specific forms without altering its technical concept or essential features. In this regard, the embodiments described above should be understood as illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as including all modifications or variations derived from the meaning and scope of the claims set forth below and their equivalents, rather than from the detailed description above.

Claims

1. A composition for alleviating and preventing cellular aging comprising TES Trioleate and Oleamide Serinol as active ingredients.

2. A composition for alleviating and preventing cell aging according to claim 1, wherein the weight ratio of the testriooleate and oleamide sulinol is 1:1 to 1:

3.

3. A composition for alleviating and preventing cell aging according to claim 1, wherein the testriooleate comprises 0.01 to 10 weight% based on the total weight of the composition.

4. A composition for alleviating and preventing cell aging according to claim 1, wherein the oleamide serinol comprises 0.01 to 10 weight% based on the total weight of the composition.

5. A composition for alleviating and preventing cell aging according to claim 1, wherein the oleamide sulinol is N-(1,3-dihydroxypropane-2-nyl)oleamide.

6. A composition for alleviating and preventing cellular aging according to claim 1, wherein the composition further comprises one or more of echinacea extract or fermented yuzu extract.

7. A composition for alleviating and preventing cell aging according to claim 6, wherein the content of the echinacea extract is 1:1 to 1:3 by weight relative to the testriooleate.

8. A composition for alleviating and preventing cell aging according to claim 6, wherein the content of the fermented yuzu extract is 1:1 to 1:5 relative to the weight of the testioleate.

9. A composition for alleviating and preventing cellular aging according to any one of claims 1 to 9, wherein the composition for alleviating and preventing cellular aging is a cenomorphic that restores the function or morphology of aging cells to the function or morphology of normal cells.

10. A composition for alleviating and preventing cell aging according to any one of claims 1 to 9, wherein the composition for alleviating and preventing cell aging is a pharmaceutical composition.

11. A composition for alleviating and preventing cell aging according to any one of claims 1 to 9, wherein the composition for alleviating and preventing cell aging is a quasi-drug composition.

12. A composition for alleviating and preventing cell aging according to any one of claims 1 to 9, wherein the composition for alleviating and preventing cell aging is a cosmetic composition.