Senotherapy composition containing camellia sinensis root extract

Abstract

To provide a senotherapy composition.SOLUTION: In one aspect, a composition of the present invention contains the tea tree (Camellia Sinensis) root extract and has an excellent senolytic effect to selectively remove senescent cells and an excellent senomorphic effect to inhibit the production of senescence associated secretory phenotype (SASP) in senescent cells.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] Disclosed herein are senotherapy compositions. [Background technology]

[0002] Senescent cells are characterized by their low reactivity and high resistance to death compared to young cells, making them less likely to die. In addition to their resistance to death, senescent cells secrete the senescence associated secretory phenotype (SASP), a complex of various aging-promoting factors, which promotes senescence and reduces the function of surrounding cells. Therefore, preventing the spread of SASP is a major challenge in preventing senescence. In addition, substances that induce the selective death of senescent cells are considered to be excellent drug candidates for preventing age-related diseases and the decline in cellular function. Most of the senescent cell-selective death inducers developed so far are drugs containing anticancer drugs that cannot be used as cosmetic ingredients due to their strong cytotoxicity, and only a few of them are effective against skin cells.

[0003] Senotherapy is a therapeutic approach that targets senescent cells, including senolytics, which selectively remove senescent cells, and senomorphics, which suppress SASP production in senescent cells. Senolytics induce apoptosis in senescent cells, and a representative example is ABT737, a Bcl-2 inhibitor. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Korean Patent Application Publication No. 10-2023-0066593 Summary of the Invention [Problem to be solved by the invention]

[0005] The present invention aims to provide a senolytic composition.

[0006] It is another object of the present invention to provide a senomorphic composition. [Means for solving the problem]

[0007] To achieve the above object, in one aspect, the present invention provides a senolytic composition comprising an extract of the root of the tea plant (Camellia Sinensis) as an active ingredient.

[0008] In another aspect, the present invention provides a senomorphic composition comprising Camellia Sinensis root extract as an active ingredient. Effect of the Invention

[0009] In one aspect, the composition of the present invention has an excellent senolytic effect of selectively removing senescent cells.

[0010] In another embodiment, the composition of the present invention has a superior senomorphic effect of inhibiting the production of Senescence Associated Secretory Phenotype (SASP) in senescent cells.

[0011] In another aspect, the composition of the present invention can improve skin elasticity and wrinkles due to the senolytic and senomorphic effects, and can also be used to prevent, improve, and treat aging-related diseases.

[0012] In another embodiment, the composition of the present invention has an excellent effect of improving the skin barrier.

[0013] In another embodiment, the composition of the present invention has an excellent skin pigmentation improving effect. [Brief description of the drawings]

[0014] [Figure 1] FIG. 1 is a diagram regarding the senolytic effect of Camellia Sinensis root extract. [Figure 2A] FIG. 1 shows an increase in apoptosis in senescent cells by treatment with tea plant root extract. [Figure 2B] FIG. 1 shows an increase in apoptosis in senescent cells by treatment with tea plant root extract. [Diagram 3] FIG. 1 shows the senomorphic effect of tea root extract, specifically, the reduction in the expression of genes related to Senescence Associated Secretory Phenotype (SASP) by treatment with tea root extract. [Figure 4A] FIG. 1 shows a graph showing a decrease in beta-galactosidase, an aging factor, by treatment with tea plant root extract. [Figure 4B] FIG. 1 shows a graph showing a decrease in beta-galactosidase, an aging factor, by treatment with tea plant root extract. [Figure 5A] FIG. 1 shows the recovery of elasticity (A), dermis (B), and epidermal function (C) of an artificial skin model by treatment with tea plant root extract. [Figure 5B] FIG. 1 shows the recovery of elasticity (A), dermis (B), and epidermal function (C) of an artificial skin model by treatment with tea plant root extract. [Figure 5C] FIG. 1 shows the recovery of elasticity (A), dermis (B), and epidermal function (C) of an artificial skin model by treatment with tea plant root extract. [Figure 6A]This figure shows the adverse effects of SASP secreted from senescent cells on surrounding keratinocytes, and the inhibition of SASP at the protein level (A) and mRNA level (B) by treatment with tea plant root extract. [Figure 6B] This figure shows the adverse effects of SASP secreted from senescent cells on surrounding keratinocytes, and the inhibition of SASP at the protein level (A) and mRNA level (B) by treatment with tea plant root extract. [Figure 7A] FIG. 1 shows the catechin (A) and amino acid (B) contents of tea plant root extract and the results of component analysis. [Figure 7B] FIG. 1 shows the catechin (A) and amino acid (B) contents of tea plant root extract and the results of component analysis. [Figure 8] FIG. 1 is a graph showing the toxicity of epigallocatechin gallate (EGCG) treatment on young fibroblasts. [Figure 9A] FIG. 1 is a graph showing the effect of treatment with tea plant root extract on improving skin pigmentation. [Figure 9B] FIG. 1 is a graph showing the effect of treatment with tea plant root extract on improving skin pigmentation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The present invention will be described in detail below. In Figures 1 to 9, GTR stands for tea plant root extract.

[0016] The term "application" as used herein means providing the composition of the present invention to a subject by any suitable method, including administration, application, absorption, ingestion, etc. In this case, the subject means any animal, such as a human, monkey, dog, goat, pig, or rat, to which the composition can be applied.

[0017] The term "cellular senescence" as used herein refers to a state in which normal cells lose the ability to divide and undergo cell cycle arrest. The cellular senescence may progress to a stage in which cell division stops, a stage in which cells that have stopped dividing secrete a senescence associated secretory phenotype (SASP) to expand senescence, and a stage in which senescent cells accumulate in tissues.

[0018] As used herein, the term "senolytics" is a compound word combining senescence and lytic, and refers to a substance that selectively removes senescent cells.

[0019] As used herein, the term "senomorphics" is a compound word combining senescence and morphic, and refers to substances that restore normal cellular levels by inhibiting the secretion of specific factors, including the inhibition of Senescence Associated Secretory Phenotype (SASP) factors.

[0020] In one aspect, the present invention relates to a senolytic composition comprising Camellia Sinensis root extract as an active ingredient.

[0021] Tea plant (Camellia Sinensis) root extract usually contains oleanane saponin as the main component (e.g., 53% of the total content). The components of tea plant root extract are completely different from those of tea plant leaf extract. The catechin content of tea plant leaf extract is about 30% by weight, whereas the catechin content of tea plant root extract is only about 0.2% by weight. This means that the effect of catechin cannot be expected from tea plant root extract.

[0022] In one exemplary embodiment, the senolytic may be a skin cell senolytic.

[0023] In one exemplary embodiment, the skin cells may be fibroblasts.

[0024] In one exemplary embodiment, the senolytic may be a senolytic caused by apoptosis.

[0025] In an exemplary embodiment, the tea plant root extract can be obtained according to a method known in the art. Specifically, the tea plant root is washed with purified water, dried in the sun or by hot air drying, and then pulverized. The powder can be prepared using an extraction solvent selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform, or a mixture of these organic solvents and water. Considering the safety of the raw materials, water or ethanol at a concentration of 30-70% can be used. The extract obtained using the solvent is extracted under reflux, filtered, and concentrated under reduced pressure at 40-50°C to obtain a dried tea plant root extract.

[0026] In one exemplary embodiment, the composition may be applied to a subject in need of restoring skin elasticity or improving wrinkles.

[0027] In one exemplary embodiment, the composition may be applied to a subject in need of a decrease in MMP-1 or an increase in procollagen in the dermis layer of the skin.

[0028] In one exemplary embodiment, the composition may be applied to a subject in need of cell generation, proliferation, or differentiation of the epidermal layer of the skin.

[0029] In one exemplary embodiment, the composition may be characterized as being applied to a subject in need of the production, delivery, or breakdown of skin pigment in the epidermal layer of the skin.

[0030] In one exemplary embodiment, the composition may be applied to a subject in need of inhibiting and breaking down the production and transmission of melanin in the epidermal layer of the skin.

[0031] In another aspect, the present invention relates to a method for selectively removing senescent cells by applying to a subject an effective amount of a tea plant root extract.

[0032] In another aspect, the present invention relates to the use of a Camellia sinensis root extract for the preparation of a senolytic composition.

[0033] In another aspect, the present invention relates to a senomorphic composition comprising tea plant root extract as an active ingredient.

[0034] In one exemplary embodiment, the senomorph may be a skin cell senomorph.

[0035] In one exemplary embodiment, the skin cells may be fibroblasts.

[0036] In an exemplary embodiment, the tea plant root extract can be obtained according to a method known in the art. Specifically, the tea plant root is washed with purified water, dried in the sun or by hot air drying, and then pulverized. The powder can be prepared using an extraction solvent selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform, or a mixture of these organic solvents and water. Considering the safety of the raw materials, water or ethanol at a concentration of 30-70% can be used. The extract obtained using the solvent is extracted under reflux, filtered, and concentrated under reduced pressure at 40-50°C to obtain a dried tea plant root extract.

[0037] In one exemplary embodiment, the composition may be applied to a subject in need of restoring skin elasticity or improving wrinkles.

[0038] In one exemplary embodiment, the composition may be applied to a subject in need of a decrease in MMP-1 or an increase in procollagen in the dermis layer of the skin.

[0039] In one exemplary embodiment, the composition may be applied to a subject in need of cell generation, proliferation, or differentiation of the epidermal layer of the skin.

[0040] In one exemplary embodiment, the composition may be characterized as being applied to a subject in need of the production, delivery, or breakdown of skin pigment in the epidermal layer of the skin.

[0041] In one exemplary embodiment, the composition may be applied to a subject in need of inhibiting and breaking down the production and transmission of melanin in the epidermal layer of the skin.

[0042] In an exemplary embodiment, the composition may be applied to a subject in need of reducing a senescence factor or a senescence associated secretory phenotype (SASP) in the skin. The senescence factor may be senescence associated beta-galactosidase (SA-β), and the SASP may be at least one selected from the group consisting of, but not limited to, IL-6, IL-8, MMP-1, and MCP-1.

[0043] In another aspect, the present invention relates to a method for reducing senescence factors or senescence associated secretory phenotype (SASP) by administering to a subject an effective amount of a Camellia sinensis root extract.

[0044] In another aspect, the present invention relates to the use of a Camellia sinensis root extract for the preparation of a senomorphic composition.

[0045] In an exemplary embodiment, the daily dosage of the active ingredient of the composition may be 10 μg / kg to 100 g / kg, more specifically, 10 mg / kg or more, 15 mg / kg or more, 25 mg / kg or more, 30 mg / kg or more, 35 mg / kg or more, 40 mg / kg or more, 45 mg / kg or more, or 50 mg / kg or more, 200 mg / kg or less, 190 mg / kg or less, 180 mg / kg or less, 170 mg / kg or less, 165 mg / kg or less, 160 mg / kg or less, 155 mg / kg or less, or 150 mg / kg or less. At the dosage, the effect of senolytics or senomorphics is excellent, and when the dosage is less than the dosage, the effect of senolytics or senomorphics is slight, and when the dosage is more than the dosage, problems such as toxicity may occur. The application may be once a day or divided into several times, for example, 2 to 24 times a day, 1 to 2 times every 3 days, 1 to 6 times a week, 1 to 10 times a week, 1 to 15 times a week, 1 to 3 times a week, or 1 to 12 times a year, but is not limited thereto.

[0046] In another aspect, the present invention relates to a composition for preventing, ameliorating or treating an aging-related disease, comprising a tea plant root extract as an active ingredient.

[0047] In an exemplary embodiment, the aging-related disease is a skin aging-related disease, such as, but not limited to, skin pruritus, dry skin, bedsores, pigmentation disorders, benign tumors, and skin cancer.

[0048] In another aspect, the present invention relates to a composition for improving skin elasticity or wrinkles, comprising a tea plant root extract as an active ingredient. Aging of skin cells, particularly a decrease in elastin and collagen produced by fibroblasts in dermal tissue, can cause wrinkles, sagging symptoms, and reduced skin elasticity, but the tea plant root extract can improve these wrinkles and skin elasticity.

[0049] In another aspect, the present invention relates to a composition for improving skin barrier, comprising a tea plant root extract as an active ingredient.

[0050] In another aspect, the present invention relates to a composition for improving skin pigmentation, comprising a Camellia sinensis root extract as an active ingredient.

[0051] In an exemplary embodiment, the composition may be at least one selected from the group consisting of a cosmetic composition, a pharmaceutical composition, and a food composition.

[0052] In an exemplary embodiment, the composition may contain 0.001 to 99.9% by weight of the active ingredient, for example, but not limited to, 0.01 to 20.0% by weight, or 0.1 to 10.0% by weight.

[0053] In an exemplary embodiment, the cosmetic composition may be prepared in the form of a general emulsion formulation and a solubilization formulation. The emulsion formulation may be a nutrient lotion, a cream, an essence, etc., and the solubilization formulation may be a softening lotion, etc. Suitable formulations may be, for example, in the form of anhydrous products in the form of solutions, gels, solids or pastes, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microgranules, or vesicular dispersions of ionic (liposome) or biionic type, creams, lotions, lotions, powders, ointments, sprays or concealing sticks, but are not limited thereto. They may also be in the form of foams or aerosol compositions further comprising a compressed propellant.

[0054] In one exemplary embodiment, the cosmetic composition may further comprise commonly used adjuvants such as fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles, or any other ingredient commonly used in cosmetic compositions.

[0055] In an exemplary embodiment, the pharmaceutical composition may be provided in any dosage form suitable for local application. For example, it may be administered orally, transdermally, intravenously, intramuscularly, or subcutaneously. For example, the pharmaceutical composition may be, but is not limited to, an injection, a solution for external application to the skin, a suspension, an emulsion, a gel, a patch, or a spray. The dosage form may be easily prepared according to a conventional method in the art, and may include, as appropriate, surfactants, excipients, hydrating agents, emulsifying agents, suspending agents, salts or buffers for adjusting osmotic pressure, coloring agents, spices, stabilizers, preservatives, preservatives, or other commonly used auxiliary agents.

[0056] In an exemplary embodiment, the amount of active ingredient in the pharmaceutical composition will depend on the subject's age, sex, weight, condition and its severity, the route of administration, or the prescriber's judgment. Determination of the appropriate dosage based on such factors is within the level of one of ordinary skill in the art.

[0057] In an exemplary embodiment, when the composition is used as an additive for health functional foods, it may be added as it is, or may be used in combination with other foods or food ingredients, and may be used appropriately according to conventional methods. The amount of the active ingredient may be appropriately determined according to each purpose of use, such as prevention, health, or treatment. The dosage form of the health functional foods may be any of powder, granule, pill, tablet, capsule, or general food or beverage forms.

[0058] In an exemplary embodiment, the type of the health functional food is not particularly limited, and examples of foods to which the composition can be added include meat, confectionery, noodles, gums, dairy products such as ice cream, various soups, soft drinks, tea, energy drinks, alcoholic beverages, and vitamin complexes, and may include all foods in the usual sense.

[0059] In an exemplary embodiment, the beverage in the health functional food may contain various flavorings, natural carbohydrates, etc. as additional ingredients, like ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrates may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the beverage according to the present invention, but is not limited thereto.

[0060] In an exemplary embodiment, in addition to the above, the health functional food according to the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloids, thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonation agents used in carbonated drinks. In addition, the health functional food according to the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used alone or in combination. The proportion of these additives is not limited, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food according to the present invention.

[0061] In another aspect, the present invention relates to a method for preventing, ameliorating or treating aging-related diseases by administering to a subject an effective amount of a tea plant root extract.

[0062] In another aspect, the present invention relates to use of a tea plant root extract for the manufacture of a composition for preventing, ameliorating or treating an aging-related disease.

[0063] In another aspect, the present invention relates to a method of improving skin elasticity or wrinkles by applying to a subject an effective amount of a tea plant root extract.

[0064] In another aspect, the present invention relates to the use of a Camellia sinensis root extract for the manufacture of a composition for improving skin elasticity or wrinkles.

[0065] In another aspect, the present invention relates to a method for improving the skin barrier by applying to a subject an effective amount of a tea plant root extract.

[0066] In another aspect, the present invention relates to the use of a Camellia sinensis root extract for the manufacture of a composition for improving skin barrier.

[0067] In another aspect, the present invention relates to a method for improving skin pigmentation by applying to a subject an effective amount of a tea plant root extract.

[0068] In another aspect, the present invention relates to the use of a Camellia sinensis root extract for the manufacture of a composition for improving skin pigmentation.

[0069] The configuration and effects of the present invention will be described in more detail below with reference to examples. However, the following examples are provided to aid in understanding the present invention, and are not intended to limit the scope and spirit of the present invention.

[0070] <Example> [Example 1] Manufacturing of Tea Root Extract Tea root extract (GTR) powder was obtained by extracting tea roots (Camellia sinensis) purchased from the Dolseong-i, Seogwang, and Hannam tea fields of the Osulrok Farm in Jeju with 70% ethanol and drying in vacuum.

[0071] [Example 2] Measurement of the degree of cell death caused by treatment with tea plant root extract Human fibroblasts were cultured in 6-well plate medium. Senescent cells were induced by culturing fibroblasts in DMEM (containing 10% FBS) medium containing doxorubicin (100 ng / mL) and insulin-like growth factor 1 (IGF-1) (100 ng / mL) for 4 days, and the same medium was treated with a positive control substance (ABT737, 1-10 μM) and tea plant root extract (50-150 ppm) and cultured for 72 hours. ABT737 is a Bcl-2 inhibitor and a representative senolytic. Young cells were treated with experimental substances in DMEM medium without inducing senescence and cultured for 72 hours.

[0072] To confirm the senolytic effect of the material treated with the tea plant root extract, the medium was replaced with a medium diluted 1 / 100 with Cell counting kit-8 (CCK-8, Dojindo), and after culturing for 1 hour, the absorbance was measured at 540 nm.

[0073] As shown in Figure 1, it was confirmed that at concentrations of 50 to 100 ppm, tea plant root extract hardly killed young cells, but only killed about 22% and 31% of senescent cells, respectively. Furthermore, it was confirmed that the effect of senescent cell death increased in a concentration-dependent manner.

[0074] [Example 3] Measurement of the degree of apoptosis in senescent cells treated with tea plant root extract Human fibroblasts were cultured in a 6-well plate medium. Senescent cells were induced by culturing fibroblasts in DMEM (containing 10% FBS) medium containing doxorubicin (100 ng / mL) and insulin-like growth factor 1 (IGF-1) (100 ng / mL) for 7 days to induce senescence, and the same medium was treated with a positive control substance (ABT737 10 μM) and tea plant root extract (150 ppm), and the cells were harvested after culturing for 24 hours. The harvested cells were treated using an apoptosis assay kit (APC annexin V / Dead cell apoptosis kit with APC annexin V and SYTOX green for flow cytometry, Invitrogen) according to the manufacturer's instructions, and the fluorescence value of each cell was measured using a FACS (BD FACSuite) device.

[0075] As shown in Figures 2A and 2B, apoptosis was slightly decreased in young cells (8.4%) compared to the negative control group (10.1%), whereas apoptosis was significantly increased in aged cells (26.6%) compared to the negative control group (18.8%).

[0076] [Example 4] Measurement of gene expression related to the Senescence Associated Secretory Phenotype (SASP) in senescent cells treated with tea root extract Human fibroblasts were cultured in 6-well plate medium. Senescent cells were induced by culturing fibroblasts in DMEM (containing 10% FBS) medium containing doxorubicin (100 ng / mL) and insulin-like growth factor 1 (IGF-1) (100 ng / mL) for 4 days, and the same medium was treated with a positive control substance (ABT737 2.5 μM) and tea plant root extract (50-150 ppm) and cultured for 72 hours. The cells were transferred to a new 6-well plate medium and cultured for another 7 days, after which the cells were harvested and genes were collected. cDNA was synthesized using 1 ug of the collected mRNA, and real-time qPCR (thermofisher) was performed to compare gene expression. The genes for IL-6 (Hs00985639_m1), IL-8 (Hs00174103_m1), and MMP-1 (Hs00899658_m1) were quantified by normalization to GAPDH (Hs02786624_g1).

[0077] As shown in Figure 3, treatment with tea root extract reduced the expression of SASP genes (IL-6, IL-8, MMP-1) that was increased by senescence induction, and the expression levels were maintained even after 7 days.

[0078] [Example 5] Measurement of senescence factor expression in senescent cells treated with tea root extract Human fibroblasts were cultured in a 6-well plate medium. Senescent cells were induced by culturing the fibroblasts in DMEM (containing 10% FBS) medium containing doxorubicin (100 ng / mL) and insulin-like growth factor 1 (IGF-1) (100 ng / mL) for 4 days, and the same medium was treated with Camellia sinensis root extract (10-150 ppm) and cultured for 72 hours, after which the cells were harvested. The harvested cells were treated for 15 minutes with medium diluted 1 / 1,000 with Spider's SPiDER-βGal (Dojindo), and the fluorescence value of each cell was measured by FACS (BD FACSuite).

[0079] As shown in FIG. 4A and FIG. 4B, when the concentration of the tea plant root extract was 50 ppm or more, the effect of reducing β-galactosidase by about 30% was confirmed.

[0080] [Example 6] Confirmation of changes in dermal layer elasticity and epidermal layer differentiation in an artificial skin model by treatment with tea plant root extract Human fibroblasts were cultured in 100 pie medium. Senescent cells were induced by culturing the fibroblasts in DMEM (containing 10% FBS) medium containing doxorubicin (100 ng / mL) and insulin-like growth factor 1 (IGF-1) (100 ng / mL) for 4 days, and the same medium was treated with tea plant root extract (100 ppm) and cultured for 72 hours. 1 × 10 fibroblasts were cultured in DMEM (containing 10% FBS) medium containing doxorubicin (100 ng / mL) and insulin-like growth factor 1 (IGF-1) (100 ng / mL). 5 The dermis layer was created by culturing the cells in collagen gel. Melanoderm (Martek), which is composed of epidermis, was purchased, and the dermis layer was artificially attached to the bottom of the cell and cultured for two weeks. After 14 days, the culture medium was collected and the expression levels of procollagen and MMP-1 were analyzed by ELISA. The cultured artificial skin was confirmed to express Ki67, K10, and Flaggrin by immunostaining.

[0081] As shown in Figure 5A, the dermal layer made from young fibroblasts had a strong pulling force on the surrounding collagen, and strong contraction was observed, whereas the dermal layer made from aged fibroblasts showed weak contraction. On the other hand, it was confirmed that the dermal layer made from aged fibroblasts treated with Camellia sinensis root extract had improved contractility. This was also confirmed by the fact that the ratio of MMP-1 and procollagen produced and secreted in the dermal layer was restored to normal, as shown in Figure 5B.

[0082] We also examined the effects on the surrounding epidermal layer by immunostaining. As shown in Figure 5C, it was confirmed that the proliferation and differentiation ability of the epidermal layer, which had been reduced by aging, was improved by treatment with tea root extract.

[0083] Furthermore, the negative effects of SASP on surrounding keratinocytes and the inhibitory effect of SASP on protein (A) and mRNA (B) levels by treatment with tea root extract were also confirmed. When the supernatant of senescent cells (old conditioned media; OCM) was treated, the skin barrier-related markers KRT10 and loricrin were decreased, and the cell damage-related factor r-H2X was increased compared to the control group (young conditioned media; YCM) (Figure 6A). However, in the case of conditioned media obtained by treating senescent cells with tea root extract, such skin barrier or cell damage was restored (Figure 6B). Specifically, p16, a senescence marker, was decreased, FLG and K10, markers related to keratinocyte differentiation and skin barrier, were increased, and SASP IL-6 and MCP-1 were decreased (Figure 6B). The decreased expression of senescence and SASP markers, and the increased expression of keratinocyte differentiation and skin barrier related markers confirmed the senomorphic and skin barrier improving effects of the tea plant root extract.

[0084] [Example 7] Content of catechins and amino acids in tea root extract and results of component analysis The tea leaf extract and tea root extract were analyzed for catechin and amino acid contents by HPLC analysis.

[0085] As a result, as shown in Figures 7A and 7B and Tables 1 and 2, it was confirmed that the contents and components of catechins and amino acids contained in tea leaf extract and tea root extract are significantly different. In tea root extract, catechins including epigallocatechin gallate (EGCG) were detected at a level of 0.2%. In other words, since tea root extract contains almost no EGCG, it is clear that the senolytic effect is not due to EGCG.

[0086] [Table 1]

[0087] [Table 2]

[0088] [Example 8] Measurement of cytotoxicity by treatment with epigallocatechin gallate (EGCG) Human fibroblasts were cultured in 6-well plate medium. Senescent cells were induced by culturing the fibroblasts in DMEM (containing 10% FBS) medium containing doxorubicin (100 ng / mL) and insulin-like growth factor 1 (IGF-1) (100 ng / mL) for 4 days, and the same medium was treated with a positive control substance (ABT737, 1-10 μM) and tea plant root extract (50-150 ppm) and cultured for 72 hours. Young cells were treated with experimental substances in DMEM medium without inducing senescence and cultured for 72 hours.

[0089] To confirm the senolytic effect of the material treated with the tea plant root extract, the medium was replaced with a medium diluted 1 / 100 with Cell counting kit-8 (CCK-8, Dojindo), and after culturing for 1 hour, the absorbance was measured at 540 nm.

[0090] As shown in FIG. 8, it was confirmed that EGCG had no effect on killing senescent cells in fibroblasts, and showed cytotoxicity only to young cells at concentrations of 50 ppm or more.

[0091] [Example 9] Ex-vivo confirmation of pigmentation improvement effect of tea plant root extract Genoskin (GENOSKIN, Toulouse, France), which was made from human tissue from the abdomen of a 46-year-old woman, was purchased. Young or senescent human fibroblasts were treated with a positive control substance (ABT737, 10 μM) and tea plant root extract (100 ppm) for 3 days, and then these fibroblasts were co-cultured with Genoskin for 7 days. Fontana-Masson staining was performed to confirm melanin pigmentation, and changes in PEML, a melanocyte marker, and tyrosinase, a protein involved in melanin synthesis, were confirmed.

[0092] As shown in Figures 9A and 9B, senescent fibroblasts increased pigmentation in ex-vivo human skin, but treatment with tea plant root extract improved the pigmentation. Immunostaining confirmed that senescent fibroblasts increased melanin production in the surrounding melanocytes, and treatment with tea plant root extract reduced the increased melanin content. Furthermore, PEML was unchanged, and tyrosinase activity was reduced by treatment with tea plant root extract.

Claims

1. A composition for senolytics; senomorphic; for preventing, improving or treating ageing-related diseases; for improving skin elasticity or wrinkles; for improving the skin barrier; or for improving skin pigmentation, comprising an extract of Camellia Sinensis root as an active ingredient.

2. The composition of claim 1 , wherein the senolytic or senomorphic is a skin cell senolytic or a skin cell senomorphic.

3. The composition of claim 2 , wherein the skin cells are fibroblasts.

4. The composition of claim 1 , wherein the senolytic is an apoptosis-inducing senolytic.

5. The composition of claim 1, wherein the composition is applied to a subject in need of reducing senescence factors or senescence associated secretory phenotype (SASP) in skin cells.

6. The senomorphic composition according to claim 5, wherein the aging factor is beta-galactosidase.

7. The senomorphic composition according to claim 5, wherein the SASP is at least one selected from the group consisting of IL-6, IL-8, MMP-1, and MCP-1.

8. The composition according to any one of claims 1 to 7, wherein the tea plant root extract is an aqueous ethanol extract of tea plant root.

9. The composition according to any one of claims 1 to 7, wherein the composition is at least one selected from the group consisting of a cosmetic composition, a pharmaceutical composition, and a food composition.

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