Topical composition for Anti-skin aging and skin cell activation, and composition for senescent cell removal and skin cell activation

Abstract

To remove senescent cells and activate skin cells, and / or to remove senescent cells to prevent, inhibit, and ameliorate skin aging phenomena such as spots, wrinkles, sagging, skin thinning, skin fragility, and dryness, and activate skin cells.SOLUTION: A composition comprises at least one plant extract selected from the group consisting of Rosaceae Rosa, Perilla frutescens var. crispa, Japanese cherry, Coffea arabica, and Arnica montana.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to an anti-skin aging and skin cell stimulating composition for external use, and a composition for removing aged cells and stimulating skin cells, etc. [Background technology]

[0002] In recent years, the phenomenon of cellular senescence, in which cell proliferation ceases permanently, has been attracting attention. For example, in analyses using genetically modified mice, p16 INK4α It has been reported that the incidence of various age-related diseases such as arteriosclerosis, cataracts, sarcopenia, and carcinogenesis was reduced by removing cells that express the gene (Non-Patent Document 1).

[0003] Cells whose proliferation has stopped permanently are called senescent cells. Senescent cells are not only inhibited in proliferation, but are also known to produce and secrete a variety of physiologically active substances, such as proteases, including extracellular matrix degrading enzymes, inflammatory cytokines, chemokines, growth factors, and exosomes. This phenomenon is known as the "senescence-associated secretory phenotype (SASP)" (Non-Patent Document 1). In the present disclosure, physiologically active substances secreted by SASP may be referred to as SASP factors.

[0004] It has been reported that SASP factors secreted from senescent cells act on neighboring cells to induce inflammatory reactions, promote or inhibit cell proliferation, induce cellular senescence, and the like. In other words, senescent cells can induce inflammatory reactions in neighboring normal (non-senescent) cells or transform normal cells into senescent cells through the secretion of SASP factors. Therefore, it is expected that inflammatory reactions and cellular senescence in surrounding cells can be suppressed by removing senescent cells that secrete SASP factors. It is known that inflammatory reactions in the skin increase the amount of melanin produced, which is one of the causes of age spots.

[0005] As mentioned above, p16 INK4αIt is known that the gene is involved in the induction of cell senescence. Non-Patent Document 2 suggests a relationship between the number of p16INK4a-positive cells in skin tissue and skin aging phenomena such as the morphological characteristics of elastic fibers and the degree of wrinkles.

[0006] It is also known that senescent cells are positive for SA-β-gal (senescence-associated beta galactsidase). In Non-Patent Document 3, it is reported that, as a result of performing SA-β-gal staining on skin samples taken from subjects of various ages, the frequency of SA-β-gal positive cells was higher in the elderly subject group aged 69 years or older than in the young subject group aged under 40 years. More specifically, SA-β-gal positive cells were not observed in the dermal samples taken from the subject group aged under 40 years, while SA-β-gal positive cells were observed in almost all of the dermal samples taken from the subject group aged 69 years or older. In addition, while SA-β-gal positive cells were observed in all of the subject group aged 69 years or older in the epidermis, SA-β-gal positive cells were observed in about half of the subject group aged under 40 years. However, in the subject group aged under 40 years, the frequency of SA-β-gal positive cells was lower in samples in which they were observed than in the subject group aged 69 years or older. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Eiji Hara, "New Aspects of Cellular Aging: Possibilities for a More Complete Understanding and Extending Healthy Lifespan," Experimental Medicine, July 2019, Vol. 37, No. 11, pp. 1728-1734 [Non-Patent Document 2] Waaijer ME et al. "P16INK4a Positive Cells in Human Skin Are Indicative of Local Elastic Fiber Morphology, Facial Wrinkling, and Perceived Age." J Gerontol A Biol Sci Med Sci. 2016 Aug;71(8):1022-1028. [Non-Patent Document 3] Dimri GP et al., "A biomarker that identifies senescent human cells in culture and in aging skin in vivo." Proc Natl Acad Sci US A. 1995 Sep 26;92(20):9363-9367. [Non-Patent Document 4] Akiko Takahashi, "Fundamentals of cellular senescence and trends in senolytics," Experimental Medicine, February 2022, Vol. 40, No. 3, pp. 364-370 Summary of the Invention [Problem to be solved by the invention]

[0008] As mentioned above, p16 INK4α It is known that genes are involved in the induction of cellular senescence, and a correlation has been suggested between the number of p16INK4a-positive cells in skin tissue and skin aging phenomena, suggesting the possibility that cellular senescence is related to skin aging phenomena such as wrinkles and sagging. As mentioned above, it has been reported that SASP factors secreted from senescent cells act on neighboring cells to induce inflammatory responses and cellular senescence, etc., so it is expected that removing senescent cells will suppress the induction of inflammatory responses and cellular senescence in surrounding cells, thereby suppressing the progression of skin aging phenomena such as age spots, wrinkles, sagging, thinning of the skin, fragility of the skin, and dryness.

[0009] In addition to removing senescent cells, it is also expected that by activating normal skin cells, cell turnover will be promoted, thereby exerting particularly effective effects in preventing, inhibiting, and improving skin aging.

[0010] In view of the above circumstances, the inventors set out to remove senescent cells and revitalize skin cells, and / or to prevent, inhibit, improve, etc., skin aging phenomena such as spots, wrinkles, sagging, thinning of the skin, fragility of the skin, and dryness, and to revitalize skin cells by removing senescent cells. [Means for solving the problem]

[0011] The present inventors have discovered that a composition containing at least one plant extract selected from the group consisting of rose, perilla, cherry, coffee tree, and arnica has the effect of removing aged cells and revitalizing normal skin cells, and have made further improvements, thereby completing the present disclosure.

[0012] The present disclosure includes, for example, the subject matter described in the following sections: Section 1. An external composition for anti-skin aging and skin cell stimulation, comprising at least one plant extract selected from the group consisting of rose, perilla, cherry blossom, coffee tree, and arnica. Section 2. A composition for removing aging cells and stimulating skin cells, comprising at least one plant extract selected from the group consisting of rose, perilla, cherry blossom, coffee tree, and arnica. Section 3. Item 3. The composition according to item 1 or 2, wherein the content of the plant extract is 0.0001 to 0.2% by mass. Section 4. Item 4. The composition according to any one of Items 1 to 3, which is used for humans aged 40 or older. Effect of the Invention

[0013] According to the technology disclosed herein, senescent cells can be removed and skin cells can be activated, and skin aging phenomena such as age spots, wrinkles, sagging skin, thinning of the skin, weakened skin, dryness, etc. can be prevented, inhibited, improved, etc. [Brief description of the drawings]

[0014] [Figure 1] The results of Test Example 1-2 are shown in the figure. In the figure, "NHEK" indicates NHEK before the procedure of Test Example 1-1 (i.e., untreated), and "SHEK" indicates cells prepared in Test Example 1-1 (i.e., senescent cells). [Diagram 2] The results of Test Example 2 for rose extract and perilla extract are shown. The incubation time after addition of the plant extract was 24 hours. The top of the bar graph indicates the average cell viability, and the error bars indicate ±standard deviation. [Diagram 3] The results of Test Example 2 for cherry extract, coffee extract, and arnica extract are shown. The incubation time after addition of the plant extracts was 48 hours. The top of the bar graph indicates the average cell viability, and the error bars indicate ± standard deviation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Each embodiment included in the present disclosure will be described in more detail below. The present disclosure preferably includes an anti-skin aging and skin cell stimulating topical composition, an aged cell removing and skin cell stimulating composition, etc., but is not limited thereto, and the present disclosure includes all that is disclosed in the present specification and that can be recognized by a person skilled in the art. Hereinafter, the anti-skin aging and skin cell stimulating topical composition, and an aged cell removing and skin cell stimulating composition included in the present disclosure may be collectively referred to as the "composition of the present disclosure."

[0016] The composition of the present disclosure contains at least one plant extract selected from the group consisting of rose, perilla, cherry, coffee tree, and arnica. In the present disclosure, each plant extract selected from the group may be collectively referred to as the "plant extract of the present disclosure."

[0017] The composition of the present disclosure containing at least one plant extract of the present disclosure can remove senescent cells and activate skin cells. Senescent cells secrete SASP factors and express p16 INK4α Senescent cells are known to have several characteristics that differ from normal cells, such as increased gene expression (Non-Patent Document 4), but in this disclosure, "senescent cells" refer to SA-β-gal-positive cells.

[0018] In the present disclosure, "removing senescent cells" refers to inhibiting the survival of senescent cells. Whether or not a certain component has a senescent cell removing effect can be confirmed by adding the component to a medium and culturing senescent cells. In other words, if the survival rate of senescent cells when cultured with the addition of the component is lower than the survival rate of senescent cells when cultured without the addition of the component, the component is determined to have a senescent cell removing effect.

[0019] In the present disclosure, "skin cell activation" refers to increasing the survival rate and / or promoting the proliferation of normal (non-senescent) skin cells. Whether or not a certain component has a skin cell activation effect can be confirmed by adding the component to a medium and culturing skin cells. In other words, if the survival rate and / or proliferation rate of skin cells when cultured with the component is higher than the survival rate and / or proliferation rate of skin cells when cultured without the component, the component is determined to have a skin cell activation effect.

[0020] As described above, senescent cells can cause inflammatory reactions in nearby normal (non-senescent) cells or change normal cells into senescent cells through the secretion of SASP factors. Therefore, it is expected that the composition of the present disclosure, which has a senescent cell removal effect, can suppress inflammatory reactions and cellular senescence caused by SASP factors. In other words, the composition of the present disclosure can also have an anti-skin aging effect.

[0021] In the present disclosure, the term "skin aging" refers to skin changes associated with aging. Specific examples of skin changes associated with aging include appearance changes such as an increase in the number of age spots, worsening of the level of age spots (size, darkness, etc.), an increase in the number of wrinkles, worsening of the level of wrinkles (depth, length, etc.), reduced elasticity (firmness), sagging, etc., and changes in skin tissues such as reduction and degeneration of collagen, elastin, etc., thinning of the skin, fragility of the skin, dryness, etc.

[0022] In the present disclosure, the term "anti-skin aging" includes preventing, inhibiting, and improving skin aging.

[0023] As mentioned above, Non-Patent Document 3 reports that, as a result of SA-β-gal staining of skin samples taken from subjects of various ages, the frequency of SA-β-gal positive cells was higher in the elderly subject group aged 69 years or older than in the young subject group aged under 40 years. That is, senescent cells tend to increase with age. Although not particularly limited, the composition of the present disclosure is preferably applied to humans aged 40 years or older, more preferably applied to humans aged 45 years or older, even more preferably applied to humans aged 50 years or older or 55 years or older, and particularly preferably applied to humans aged 60 years or older or 65 years or older. The lower limit of the range may be 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, or 70 years old.

[0024] Preferred examples of applications of the composition of the present disclosure include people who wish to prevent skin aging, people who wish to inhibit skin aging, people who wish to improve skin aging, people who are concerned about age spots, people who want to prevent age spots, people who want to inhibit the progression of age spots, people who want to improve age spots, people who are concerned about wrinkles, people who want to prevent wrinkles, people who want to inhibit the progression of wrinkles, people who want to improve wrinkles, people who are concerned about sagging, people who want to prevent sagging, people who want to inhibit the progression of sagging, people who want to improve sagging, people who are concerned about disrupted turnover, people who want to improve disrupted turnover, people who are interested in anti-skin aging, and the like.

[0025] Preparation of the Plant Extracts of the Present Disclosure In the present disclosure, the term "plant extract" preferably includes any of an extract obtained by an extraction operation using a plant, a dilution of the extract, a concentrate of the extract, and a dried product obtained by drying the extract. The extract may be appropriately fractionated and / or purified by a conventional method within a range in which the desired effect is obtained. That is, the term "plant extract" in the present disclosure also includes fractions, crude products, and purified products of the extract.

[0026] The part of the plant extract of the present disclosure is not particularly limited as long as it can obtain the desired effect.The part of the plant extract can be, for example, root, stem, leaf, flower, bud, rhizome, seed, fruit, peel, branch, trunk, bark, etc.The part of the plant extract can be a single part or a mixture of multiple parts, or the whole plant.The part of the plant extract can be appropriately selected according to the characteristics of the plant.

[0027] The method for preparing the plant extract of the present disclosure is not particularly limited as long as the desired effect can be obtained. For example, the extraction solvent, the amount of the extraction solvent used, the extraction time, the extraction temperature, etc. can be appropriately selected according to the type and part of the plant to be subjected to the extraction operation. In addition, the plant to be subjected to the extraction operation may be shredded or crushed with or without drying.

[0028] Examples of the extraction solvent include water, methanol, ethanol, propanol, isopropanol, 1,3-butylene glycol, propylene glycol, glycerin, benzene, toluene, xylene, ethyl ether, methyl ethyl ketone, methyl isobutyl ketone, dichloromethane, dichloroethane, chloroform, ethyl acetate, propyl acetate, butyl acetate, and acetone. A mixture of two or more solvents may be used as the extraction solvent. In this case, the mixing ratio may be appropriately adjusted depending on the characteristics and condition of the plant to be subjected to the extraction operation. The extraction solvent is preferably one or more selected from the group consisting of water, ethanol, 1,3-butylene glycol, propylene glycol, and glycerin.

[0029] The extraction time may be, for example, about 30 minutes to 10 days. The upper or lower limit of the extraction time may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or 1, 2, 3, 4, 5, 6, 7, 8, or 9 days. The extraction time may be appropriately adjusted depending on the characteristics and condition of the plant to be subjected to the extraction operation.

[0030] The extraction temperature may be, for example, about 4° C. to 120° C. The upper or lower limit of the extraction temperature may be 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, or 115° C. The extraction temperature may be appropriately adjusted depending on the characteristics and condition of the plant to be subjected to the extraction operation.

[0031] More specifically, the extraction method may include immersing the plant in an extraction solvent, leaving it to stand or stirring appropriately for about 30 minutes to 10 days to dissolve the soluble components in the solvent, and then filtering to remove solids. The extraction may also be heated appropriately.

[0032] When the extract is purified, for example, it may be performed using activated carbon, porous resin, ion exchange resin, adsorbent, high performance liquid chromatography (HPLC), etc. Two or more purification methods may be combined.

[0033] Additionally, as described below, commercially available products may also be used as the plant extracts of the present disclosure.

[0034] Content of Plant Extracts of the Present Disclosure In the composition of the present disclosure, the content of the plant extract of the present disclosure is not particularly limited, but may be, for example, 0.00001 to 1 mass%, preferably 0.00005 to 0.5 mass%, more preferably 0.0001 to 0.2 mass%, even more preferably 0.0001 to 0.1 mass%, and particularly preferably 0.0001 to 0.05 mass%. The upper or lower limit of the range may be, for example, 0.00005, 0.0001, 0.00015, 0.0002, 0.00025, 0.0003, 0.00035, 0.0004, 0.00045, 0.0005, 0.00055, 0.0006, 0.00065, 0.0007, 0.00075, 0.0008, 0.00085, 0.0009, 0.00095, 0.001, 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045, 0 It may be 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1% by weight.

[0035] In addition, when a commercially available product is used as the plant extract of the present disclosure, the commercially available product may contain a solvent such as water, a preservative, etc. in addition to the plant extract. In this case, the content of the plant extract in the composition of the present disclosure is calculated from the content of the plant extract in the commercially available product disclosed by the manufacturer of the commercially available product and the content of the commercially available product in the composition of the present disclosure. For example, if the content of the plant extract in the commercially available product is 10% and the content of the commercially available product in the composition of the present disclosure is 10%, the content of the plant extract in the composition of the present disclosure is calculated to be 1%.

[0036] When a composition of the present disclosure contains two or more types of plant extracts of the present disclosure, the "content of the plant extract of the present disclosure" refers to the total content of each extract.

[0037] Each plant extract of the present disclosure will be described below.

[0038] Rose Extract Rose (Rosa rosa) is a general term for plants of the genus Rosaceae. Examples of plants of the genus Rosaceae include Rosa centifolia, Rosa canina, Rosa damascena, Rosa gallica, Rosa multiflora, and Rosa rugosa. Rosa centifolia, Rosa canina, Rosa damascena, Rosa gallica, Rosa multiflora, and Rosa rugosa can all be preferably used as raw materials for the rose extract of the present disclosure. The rose extract may be used alone or in combination of two or more. The part of the rose to be extracted is not particularly limited, but it is preferable to include the flower part. The composition of the present disclosure preferably contains a rose extract made from Rosa centifolia, especially from the rose.

[0039] In the present disclosure, the rose extract may be one obtained by extraction from roses using the above-mentioned method, or a commercially available product containing rose extract may be used. Rose extract is contained in commercially available products such as "Falcorex Rose P (Ichimaru Falcos)", "Rose Extract S (Koei Kogyo)", "Rose Extract BG (Maruzen Pharmaceuticals)", "Falcorex Nobara B (Ichimaru Falcos)", and "Nobara Extract (Koei Kogyo)". When the composition of the present disclosure contains rose extract, the content is not particularly limited and can be appropriately adjusted with reference to the above-mentioned "Content of Plant Extract of the Present Disclosure", and among them, it is preferably 0.0001 to 0.05% by mass, more preferably 0.0005 to 0.02% by mass, and even more preferably 0.001 to 0.01% by mass.

[0040] Perilla extract Perilla frutescens var. crispa is an annual herb of the Lamiaceae family. In the present disclosure, the perilla extract may be one obtained by extraction from perilla using the above-mentioned method, or a commercially available product containing perilla extract may be used. There are no particular limitations on the part of the perilla that may be extracted, but it is preferable that the extract contains the leaves. Perilla extract is contained in commercially available products such as "Falcolex Perilla HB (Ichimaru Falcos Co., Ltd.)", "Perilla Extract (Koei Kogyo Co., Ltd.)", "Perilla Extract BG (Maruzen Pharmaceutical Co., Ltd.)", "Perilla Extract-BG (Yamada Yaken Co., Ltd.)" and "Perilla Fruit Extract-LOGC (Oryza Oil & Fat Chemical Co., Ltd.)".

[0041] When the composition of the present disclosure contains a perilla extract, its content is not particularly limited and can be appropriately adjusted with reference to the above-mentioned "Content of plant extract of the present disclosure," but it is preferably 0.0001 to 0.01 mass%, more preferably 0.0005 to 0.005 mass%, and even more preferably 0.0008 to 0.002 mass%.

[0042] Cherry Blossom Extract Cherry blossoms are deciduous tall or shrub trees of the genus Prunus in the subfamily Prunus of the family Rosaceae. There are many varieties of cherry blossoms, including Somei-Yoshino, Yamazakura, Oshimazakura, Ooyamazakura, Edohigashi, Mamezakura, Miyamazakura, Kasumizakura, Kohigan, Satozakura, and Kanzakura. In the present disclosure, the variety of cherry blossoms is not particularly limited, but Somei-Yoshino is particularly preferred. In the present disclosure, the cherry blossom extract may be obtained by extracting cherry blossoms using the above-mentioned method, or a commercially available product containing cherry blossom extract may be used. The part of the cherry blossom to be extracted is not particularly limited, but it is preferable to include the leaves. The cherry blossom extract is contained in commercially available products such as "Sakura Extract B (Ichimaru Falcos Co., Ltd.)".

[0043] When the composition of the present disclosure contains cherry blossom extract, its content is not particularly limited and can be appropriately adjusted with reference to the above-mentioned "Content of plant extract of the present disclosure," but among these, it is preferably 0.0001 to 0.1 mass%, more preferably 0.001 to 0.05 mass%, and even more preferably 0.002 to 0.005 mass%.

[0044] Coffee extract Coffee is a general term for plants belonging to the genus Coffea in the family Rubiaceae. There are various cultivars of coffee, such as Coffee arabica and Coffee robusta. In the present disclosure, the variety of coffee is not particularly limited, but Coffea arabica is particularly preferred. In the present disclosure, the coffee extract may be one obtained by extraction from the coffee tree using the above-mentioned method, or a commercially available product containing a coffee extract may be used. There is no particular limit to the part of the coffee tree that is extracted, but it is preferable that the part contains the seeds. Coffee extract is contained in commercially available products such as "Cafe Noage (Ichimaru Falcos)" and "Coffee Extract-BG (Yamada Yaken Co., Ltd.)".

[0045] When the composition of the present disclosure contains a Coffea plant extract, its content is not particularly limited and can be adjusted as appropriate with reference to the above-mentioned "Content of plant extract of the present disclosure," but it is preferably 0.0001 to 0.005 mass%, more preferably 0.0003 to 0.001 mass%, and even more preferably 0.0005 to 0.0008 mass%.

[0046] Arnica Extract Arnica montana is a plant of the Asteraceae family. In the present disclosure, the arnica extract may be obtained by extracting arnica using the above-mentioned method, or a commercially available product containing arnica extract may be used. The part of arnica to be extracted is not particularly limited, but it is preferable to include the flower part. Arnica extract is contained in commercially available products such as "Adepurun (Ichimaru Pharcos Co., Ltd.)", "Arnica Extract (Koei Kogyo Co., Ltd.)" and "Arnica Extract BG (Maruzen Pharmaceutical Co., Ltd.)".

[0047] When the composition of the present disclosure contains arnica extract, its content is not particularly limited and can be appropriately adjusted with reference to the above-mentioned "Content of plant extract of the present disclosure," but among these, it is preferably 0.00001 to 0.005 mass%, more preferably 0.00005 to 0.002 mass%, even more preferably 0.0001 to 0.001 mass%, and particularly preferably 0.0002 to 0.0008 mass%.

[0048] Preparation of the compositions of the present disclosure The composition of the present disclosure may contain known components that can be contained in a composition for external use, as long as the effect of the present disclosure is not impaired. Examples of such known components include water, moisturizers, oils, surfactants, thickeners, film-forming agents, preservatives, pH adjusters, chelating agents, antioxidants, colorants, fragrances, refreshing agents, UV absorbers, various powders, other active ingredients, plant extracts other than the plant extracts of the present disclosure, etc. The composition of the present disclosure may contain one or more of these known components at will.

[0049] Examples of the moisturizing agent include sugars such as trehalose, lactulose, maltitol, etc., sugar alcohols such as sorbitol, mannitol, maltitol, etc., and polyhydric alcohols such as propanediol, diethylene glycol, polyethylene glycol, 1,3-butylene glycol, glycerin, diglycerin, etc. The moisturizing agents can be used alone or in combination of two or more kinds.

[0050] Examples of oils include vegetable oils, triglycerides, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, silicone oils, etc. The oils may be used alone or in combination of two or more.

[0051] Examples of the surfactant include anionic surfactants such as polyoxyethylene alkyl ether sulfates, alkyl sulfate ester salts, alkyl amide ether sulfates, polyoxyethylene alkyl amide ether sulfates, α-olefin sulfonates, alkyl sulfosuccinates, polyoxyethylene alkyl ether acetates, alkyl phosphates, polyoxyethylene alkyl ether phosphates, higher fatty acid salts, N-acyl amino acid salts, N-acyl isethionates, and N-acyl methyl taurates; Cationic surfactants such as quaternary ammonium salts; Amphoteric surfactants such as amidopropyl betaine type, amidoamine oxide type, sulfobetaine type, imidazoline type, alkyl betaine type, etc. Examples of nonionic surfactants include polyoxyalkylene alkyl ethers, polyoxyalkylene glycols, polyoxyalkylene fatty acid esters, polyoxyalkylene sorbitan fatty acid esters, polyoxyalkylene sorbitol fatty acid esters, polyoxyalkylene glycerin fatty acid esters, polyoxyalkylene fatty acid amides, polyoxyalkylene glycol fatty acid esters, polyoxyalkylene castor oil derivatives, polyoxyalkylene hydrogenated castor oil derivatives, polyglycerin fatty acid esters, sorbitan fatty acid esters, sorbitol fatty acid esters, sucrose fatty acid esters, alkylene glycol fatty acid esters, alkyl polyglycosides, and fatty acid alkanolamides.

[0052] Examples of preservatives include parabens such as methylparaben, ethylparaben, propylparaben, and butylparaben, sodium benzoate, phenoxyethanol, and alkyldiaminoethylglycine hydrochloride.

[0053] Examples of pH adjusters include citric acid, phosphoric acid, malic acid, pyrophosphoric acid, lactic acid, tartaric acid, phytic acid, glycerophosphoric acid, acetic acid, nitric acid, or chemically acceptable salts thereof, sodium hydroxide, and the like.

[0054] Examples of the chelating agent include edetic acid, pentetic acid, metaphosphoric acid, gluconic acid, etidronic acid, and chemically acceptable salts thereof.

[0055] Examples of colorants include legal dyes such as Blue No. 1, Yellow No. 4, Red No. 202, and Green No. 3.

[0056] Examples of the fragrance include menthol, anethole, carvone, eugenol, limonene, wintergreen, clove oil, rosemary oil, lemon oil, orange oil, ocimene oil, citronellol, and methyl eugenol.

[0057] Examples of cooling agents include menthol, spearmint oil, camphor, thymol, methyl salicylate, and the like.

[0058] The composition of the present disclosure may be a pharmaceutical composition or a cosmetic composition. For example, it may be a skin care agent, a hair care agent, a bath agent, etc. More specifically, it may be a skin lotion, a lotion, a beauty essence, a milky lotion, a cream, an ointment, a paste, a gel, a pack, a compress, a spray, a shampoo, a conditioner, a hair growth agent, a scalp lotion, etc.

[0059] The form of the composition of the present disclosure is not particularly limited, and may be, for example, a liquid, cream, gel, paste, semi-solid, solid, powder, or the like.

[0060] The composition of the present disclosure can be prepared by a known method or a method that a person skilled in the art can easily think of from a known method. For example, it can be prepared by mixing the plant extract of the present disclosure and other components. Mixing can be performed using a known stirrer (e.g., a dispersing mixer, etc.).

[0061] In addition, in this specification, the term "comprising" includes "essentially consisting of" and "consisting of" in addition to "containing." In addition, the present disclosure includes all arbitrary combinations of the constituent elements described in this specification. EXAMPLES

[0062] Hereinafter, the embodiments of the present disclosure will be described more specifically with reference to examples, but the embodiments of the present disclosure are not limited to the following examples.

[0063] Test Example 1. Preparation of senescent cells 1-1. Normal human epidermal keratinocytes (NHEK, Lifeline Cell Technology) were used to induce cellular senescence by a known method. Specifically, cellular senescence was induced by the following method. Unless otherwise specified below, the cells were cultured at 37°C CO 2 The experiment was carried out in a 5 v / v% incubator. (1) In a 96-well plate, 5.0 × 10 NHEK cells were cultured. 3 The cells were seeded at 1000 cells / well and cultured for 1 day in HuMedia™-KG2 (Cat. No. KK-2150S, Kurabo) culture medium. (2) After removing the medium, the medium was replaced with HuMedia-KB2 (Cat. No. KK-2350S, Kurabo) containing 1.5 μM Palbociclib (Cat. No. PZ0383, Merck) and 1 v / v% dimethyl sulfoxide (DMSO), and the cells were cultured for 24 hours. (3) The medium was removed and replaced with HuMedia-KB2 containing 1.5 μM Palbociclib, 9 μM Nutlin-3a (Cat. No. SML0580, Merck) and 1 v / v% DMSO, and cultured for 8 hours. (4) The medium was removed and replaced with HuMedia-KB2 containing Nutlin-3a 9 μM, BI-2536 (Cat. No. HY-50698, MedChemExpress) 500 nM, and DMSO 1 v / v% and cultured for 1.5 days. (5) The medium was removed and replaced with HuMedia-KB2 containing 500 nM BI-2536 and 1 v / v% DMSO, and cultured for 3 days.

[0064] 1-2. The cells prepared in Test Example 1-1 and the NHEK before the procedure in Test Example 1-1 (ie, untreated) were each subjected to SA-β-gal staining by the following method. (1) The medium in the wells was aspirated and washed twice with PBS. (2) 1× Fixing Solution included in the Cellular Senescence Assay Kit (Cat. No. CBA-230, CELL BIOLABS) was added, and the cells were fixed at room temperature for 5 minutes. (3) The fixing solution was removed, and the sections were washed three times with PBS. (4) The Cell Staining Working Solution included with the Cellular Senescence Assay Kit was added until the cells were completely submerged. (5) The reaction was allowed to proceed overnight in a 37°C incubator, protected from light. (6) The Cell Staining Working Solution was removed, and the cells were washed twice with PBS, and the wells were filled with PBS. (7) Observation was performed under an optical microscope to confirm whether the cells were stained blue. The micrograph is shown in Figure 1.

[0065] 1-3. As a result of Test Example 1-2, as shown in FIG. 1, almost all the cells prepared in Test Example 1-1 were positive for SA-β-gal staining, whereas no cells positive for SA-β-gal staining were observed in the NHEK before the operation of Test Example 1-1. That is, it was confirmed that cellular senescence was induced by the method described in Test Example 1-1, and normal NHEK became senescent cells. Therefore, the cells prepared by the method described in Test Example 1-1 were used as senescent cells in the following experiments. In this Example, the senescent cells prepared by the method described in Test Example 1-1 may be referred to as SHEK.

[0066] Test Example 2. Senescent cell elimination test 2-1. SHEK were cultured with or without the addition of each plant extract shown in Table 1 to the medium. The commercially available products shown in Table 2 were used as the plant extracts shown in Table 1. Each commercially available product shown in Table 2 contains a solvent and the like in addition to the plant extract. The concentrations (w / w%) shown in Table 1 are values ​​calculated from the content of the plant extract in the commercially available product and the concentration of the commercially available product added to the medium. In other words, the concentrations shown in Table 1 indicate the concentration of the plant extract in the medium (sometimes referred to as "net concentration" in the present disclosure), rather than the concentration of the commercially available product in the medium.

[0067] [Table 1]

[0068] [Table 2]

[0069] A specific method for culturing SHEK is shown below. (1) The medium was removed from the wells containing the SHEK cells obtained in Test Example 1-1, and the cells were washed twice with PBS. (2)-i 100 μL of HuMedia-KB2 containing rose extract or perilla extract at the concentrations shown in Table 1, or HuMedia-KB2 containing no plant extract, was added to each well and cultured for 24 hours. (2)-ii 100 μL of HuMedia-KB2 containing cherry extract, coffee extract, or arnica extract at the concentrations shown in Table 1, or HuMedia-KB2 without any plant extract, was added to each well and cultured for 48 hours.

[0070] 2-2. The viability of SHEK cells cultured in Test Example 2-1 was measured using Premix WST-1 Cell Proliferation Assay System (Cat. No. MK400, TaKaRa Bio, hereinafter sometimes referred to as "WST-1 Premix"). WST-1 is a reagent for quantifying cell viability by colorimetric measurement. It is based on the conversion of tetrazolium salt (WST-1) to formazan dye by mitochondrial dehydrogenase in living cells. If the number of living cells increases, the overall activity of mitochondrial dehydrogenase in the sample increases, and this increase in enzyme activity leads to an increase in the production of formazan dye, so that the amount of formazan dye present and the number of metabolically active cells in the medium show a linear correlation.

[0071] Specifically, the survival rate was measured by the following method. (1) The medium was removed from the wells containing the SHEK cells cultured in Test Example 2-1, and the wells were washed once with PBS. (2) 100 μL of HuMedia-KB2 containing 10 v / v% WST-1 premix was added to each well and incubated for 2 hours and 30 minutes. (3) The absorbance at 450 nm of each well was measured using an xMark™ Microplate Spectrophotometer (BIO RAD). (4) From the measured absorbance at 450 nm, the cell viability when each plant extract was added was calculated according to the following formula, with the cell viability when no plant extract was added (control) being set at 1. As mentioned above, there is a linear correlation between the amount of formazan dye that absorbs at 450 nm and the number of metabolically active cells (i.e., surviving cells). Cell viability = absorbance at 450 nm with plant extract added / absorbance at 450 nm without plant extract added (control) The test was performed with n = 5. The results are shown in Figures 2 and 3 and Tables 3 and 4. The tops of the bars indicate the average values, and the error bars indicate ±standard deviation.

[0072] 2-3. Non-senescence-induced NHEKs were also cultured with or without the addition of each plant extract shown in Table 1. Specifically, NHEKs were cultured by the following method. (1) In a 96-well plate, add 1.0 × 10 NHEK cells. 4 The cells were seeded at 1000 cells / well and cultured for 1 day using HuMedia-KG2 medium. (2)-i 100 μL of HuMedia-KB2 containing rose extract or perilla extract at the concentrations shown in Table 1, or HuMedia-KB2 containing no plant extract, was added to each well and cultured for 24 hours. (2)-ii 100 μL of HuMedia-KB2 containing cherry extract, coffee extract, or arnica extract at the concentrations shown in Table 1, or HuMedia-KB2 without any plant extract, was added to each well and cultured for 48 hours.

[0073] 2-4. The survival rate of the NHEKs cultured in Test Example 2-3 was also measured using WST-1 premix in the same manner as in Test Example 2-2. Specifically, the survival rate was measured by the following method. (1) The medium was removed from the wells containing the NHEKs cultured in Test Example 2-3, and the wells were washed once with PBS. (2) 100 μL of HuMedia-KB2 containing 10 v / v% WST-1 premix was added to each well and incubated for 2 hours and 30 minutes. (3) The absorbance of each well was measured at 450 nm using an xMark Microplate Spectrophotometer. (4) From the measured absorbance at 450 nm, the cell viability when each plant extract was added was calculated based on the following formula, with the cell viability when no plant extract was added (control) being set at 1. Cell viability = absorbance at 450 nm with plant extract added / absorbance at 450 nm without plant extract added (control) The test was performed with n=5. The results are shown in Figures 2 and 3 and Tables 3 and 4. In Figures 2 and 3, the tops of the bars indicate the average values, and the error bars indicate ±standard deviation.

[0074] [Table 3]

[0075] [Table 4]

[0076] 2-5. Discussion As shown in Figures 2 and 3 and Tables 3 and 4, when each plant extract was added to the culture medium of SHEK, the cell viability was lower than when each plant extract was not added. These results suggest that each plant extract has an effect of removing senescent cells.

[0077] Furthermore, when each plant extract was added, the cell viability was lower in SHEK than in NHEK, suggesting that each plant extract can selectively induce cell death in senescent cells.

[0078] Furthermore, when each plant extract was added to the culture medium for NHEK, the cell viability was higher than when each plant extract was not added. Here, NHEK retained the proliferation ability, and the amount of formazan pigment measured in Test Example 2 and the number of live cells in the medium showed a linear correlation. From this, it is considered that each plant extract may have promoted the proliferation of NHEK in addition to increasing the survival rate of NHEK. In other words, it was suggested that each plant extract not only has an effect of removing senescent cells, but also has an effect of promoting the proliferation of normal skin cells (i.e., a skin cell activation effect).

Claims

1. An external composition for anti-skin aging and skin cell stimulation, comprising at least one plant extract selected from the group consisting of rose, perilla, cherry blossom, coffee tree, and arnica.

2. A composition for removing aging cells and stimulating skin cells, comprising at least one plant extract selected from the group consisting of rose, perilla, cherry blossom, coffee tree, and arnica.

3. The composition according to claim 1 or 2, wherein the content of the plant extract is 0.0001 to 0.2% by mass.

4. The composition according to claim 1 or 2, which is used for humans aged 40 years or older.

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