Composition containing sesamin and ergothioneine or its salt

Abstract

The present invention aims to provide a composition that can effectively suppress cellular senescence. [Solution] A composition is provided that contains one or more types of sesamins and ergothioneine or a salt thereof, wherein the ratio of the weight of sesamins to the weight of ergothioneine or a salt thereof in terms of ergothioneine (sesamins / ergothioneine) is 0.05 to 50. Furthermore, a composition is provided that bears one or more of the following expressions selected from the group consisting of "inhibits cellular aging," "inhibits the formation of senescent cells," "prevents cellular aging," "prevents the formation of senescent cells," "inhibits the increase of senescent cells," "suppresses the accumulation of senescent cells," "delays cellular aging," "delays the progression of cellular aging," "inhibits the induction of cellular aging," "increases the proportion of young cells," "extends healthy lifespan," "healthy longevity," "young cells," "whole body youthful," and "youthful appearance."

Description

Technical Field

[0001] The present invention relates to a composition containing one or more sesamin compounds, and ergothioneine or a salt thereof. The present invention also relates to a method for inhibiting cellular senescence, and the like. Background Art

[0002] Cellular senescence refers to a state in which cell proliferation is irreversibly stopped due to causes such as genetic damage. For example, normal somatic cells of mammals such as humans have a limit on the number of divisions possible, and after repeating cell division a certain number of times, cell division is irreversibly stopped.

[0003] Unlike apoptosis, cells that have undergone cellular senescence (senescent cells) do not die on their own. For this reason, senescent cells accumulate in tissues and organs with aging (Non-Patent Document 1). Senescent cells not only have their cell cycle arrested, but also exhibit reduced function as cells. In addition, it is believed that senescent cells damage surrounding normal cells through physiologically active factors secreted from the cells, causing aging at the individual level such as functional decline of organs. Conversely, it has been reported that suppressing the accumulation of senescent cells extends lifespan (Non-Patent Document 2). Due to these properties of senescent cells, there is growing interest in inhibiting cellular senescence.

[0004] Sesamin compounds are one type of major lignan compounds contained in sesame, and anti-fatigue effects and the like have been reported (Patent Document 1). Ergothioneine is an amino acid contained in mushrooms and the like, and the L-form exists naturally. With regard to ergothioneine, physiological actions such as the effect of promoting production of immune response-activated cytokines have been reported (Patent Document 2). Prior Art Documents Patent Documents

[0005] Patent Document 1 International Publication No. WO2008 / 126587 Patent Document 2 Japanese Patent Publication No. 2017-218431 [Non-patent literature]

[0006] [Non-Patent Document 1] Herbig U, Ferreira M, Condel L, Carey D, Sedivy JM. “Cellular Senescence in Aging Primates.” Science. 2006;311(5765):1257-1257. [Non-Patent Document 2] Baker, Darren J., et al. “Naturally occurring p16 Ink4a-positive cells shorten healthy lifespan.” Nature 530.7589 (2016): 184-189. [Overview of the project] [Problems that the invention aims to solve]

[0007] However, the effects of compositions containing sesamins and ergothioneine on cellular senescence have not been reported to date.

[0008] The object of the present invention is to provide a composition that can effectively suppress cellular senescence. [Means for solving the problem]

[0009] As a result of diligent research to solve the above problems, the inventors have found that using one or more compounds of the sesamin family in combination with ergothioneine or a salt thereof can effectively suppress cellular senescence.

[0010] The present invention includes the following compositions, etc. [1] A composition containing one or more types of sesamins and ergothioneine or a salt thereof, wherein the ratio of the weight of sesamins to the weight of ergothioneine or a salt thereof in terms of ergothioneine (sesamins / ergothioneine) is 0.05 to 50. [2] The composition according to [1] above, wherein the sesamins are sesamin and / or episesamin. [3] The composition described in [1] or [2] above, which is an oral composition. [4] A composition described in any of [1] to [3] above, which is a food or beverage. [5] The composition described in [1] or [2] above, which is a cosmetic. [6] A composition according to any one of [1] to [5] above, used to suppress cellular senescence. [7] An anti-aging composition, as described in any of [1] to [6] above. [8] A composition according to any one of [1] to [7] above, which is marked with one or more of the following expressions selected from the group consisting of "inhibits cellular senescence," "inhibits the formation of senescent cells," "prevents cellular senescence," "prevents the formation of senescent cells," "inhibits the increase of senescent cells," "suppresses the accumulation of senescent cells," "delays cellular senescence," "delays the progression of cellular senescence," "inhibits the induction of cellular senescence," "increases the proportion of young cells," "extends healthy lifespan," "healthy longevity," "young cells," "youthful overall," and "youthful appearance." [9] A method for suppressing cellular senescence, comprising administering any of the compositions described in [1] to [8] above.

[10] Use of any of the compositions described in [1] to [8] above for the purpose of suppressing cellular senescence. [Effects of the Invention]

[0011] According to the present invention, it is possible to provide a composition that can effectively suppress cellular senescence. [Brief explanation of the drawing]

[0012] [Figure 1] Figure 1 is a graph showing the results of an investigation into the cellular senescence inhibitory effects of a sesamin / episesamin mixture and ergothioneine. [Figure 2] Figure 2 is a graph showing the results of investigating the cellular senescence inhibitory effect of a sesamin-episesamin mixture and ergothioneine. [Figure 3] Figure 3 is a graph showing the results of investigating the lifespan extension effect of a sesamin-episesamin mixture and ergothioneine in yeast. MODE FOR CARRYING OUT THE INVENTION

[0013] The composition of the present invention is a composition containing one or more sesamin compounds, and ergothioneine or a salt thereof, wherein the ratio of the weight of the sesamin compounds to the weight of ergothioneine or a salt thereof in terms of ergothioneine (sesamin compounds / ergothioneine) is 0.05 to 50.

[0014] In the present invention, sesamin compounds is a general term for compounds including sesamin and analogs thereof. Sesamin is one of the main lignan compounds contained in sesame. Examples of sesamin analogs include episesamin, and dioxabicyclo[3.3.0]octane derivatives described in, for example, Japanese Patent Application Laid-Open No. 4-9331. As the one or more sesamin compounds, one of these compounds may be used alone, or two or more thereof may be used. Specific examples of sesamin compounds include sesamin, episesamin, sesaminol, episesaminol, sesamol, and sesamorin, and their stereoisomers or racemates can be used alone or as a mixture. In the present invention, as one or more sesamin compounds, sesamin and / or episesamin is preferred, and sesamin and episesamin are more preferred. When sesamin and episesamin are used, their ratio is not particularly limited. For example, the weight ratio of sesamin:episesamin is preferably 1:0.1 to 1:9, more preferably 1:0.3 to 1:3, and still more preferably 1:0.5 to 1:2.

[0015] The sesamins used in the present invention are not limited at all by their form, production method, or the like. For example, in the case of sesamin, an extract containing sesamin (referred to as a sesamin extract or a purified sesamin product) extracted from sesame oil by a known method (for example, the method described in Japanese Patent Application Laid-Open No. Hei 4-9331) can be used. Commercially available sesame oil (liquid) can also be used as it is. However, when sesame oil is used, the flavor peculiar to sesame oil may be organoleptically evaluated as unfavorable, so it is preferable to use a tasteless and odorless sesamin extract (or purified sesamin product) extracted from sesame oil. In addition, when sesame oil is used, the content of sesamin is low, so when attempting to blend a preferred amount of sesamin, the volume per unit dose of the formulated composition becomes excessively large, which may cause inconvenience in intake. In particular, when formulated for oral intake, the preparation (tablet, capsule, etc.) becomes excessively large, which impairs intake. Therefore, it is also preferable to use a sesamin extract (or purified sesamin product) from sesame oil from the viewpoint that a small intake is sufficient. Sesamins can also be obtained by synthesis. As for the method, for example, sesamin and episesamin can be synthesized by the method of Beroza et al. (J. Am. Chem. Soc., 78, 1242 (1956)).

[0016] Ergothioneine is a type of sulfur-containing amino acid. In the present invention, ergothioneine is preferably L-ergothioneine. Salts of ergothioneine are not particularly limited as long as they are pharmacologically acceptable salts or salts acceptable for foods and drinks, and may be either acid salts or basic salts. Examples of acid salts include inorganic acid salts such as hydrochloride, sulfate, nitrate, and phosphate; organic acid salts such as acetate, citrate, maleate, malate, oxalate, lactate, succinate, fumarate, and propionate. Examples of basic salts include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt.

[0017] Ergothioneine or its salts are not limited in any way by their form or manufacturing method. Ergothioneine or its salts may be chemically synthesized or extracted and purified from natural sources. L-ergothioneine is abundant in the golden / yellow oyster mushroom (Pleurotus cornucopiae var. citrinopileatus), a mushroom belonging to the genus Pleurotus in the family Pleurotaceae. L-ergothioneine is also found in mushrooms such as white button mushrooms, crimson mushrooms, portabella mushrooms, oyster mushrooms (Pleurotus ostreatus), shiitake mushrooms (Lentinula edodes), maitake mushrooms (Grifola Frondosa), lychee mushrooms (Ganoderma lucidum), lion's mane mushrooms (Hericium erinaceus), willow mushrooms (Agrocybe aegerita), chanterelle mushrooms (Cantharellus cibarius), porcini mushrooms (Boletus edulis), and morel mushrooms (Morchella esculenta). When obtaining L-ergothioneine from natural sources, extraction from Pleurotus ostreatus is preferable. Ergothioneine or its salts can also be produced by microbial fermentation. Ergothioneine or its salts may be isolated.

[0018] In the composition of the present invention, the ratio of the weight of sesamins to the weight of ergothioneine or its salt in terms of ergothioneine (sesamins / ergothioneine) is 0.05 to 50. When the ratio of the weight of sesamins to ergothioneine or its salt (in terms of ergothioneine) (sesamins / ergothioneine) is within the above range, cellular senescence can be suppressed more effectively than when sesamins are used alone or when ergothioneine or its salt is used alone. As a result, the composition of the present invention can exert the above effects even at low doses. Since reducing the dosage leads to ease of administration, it is possible to obtain a composition that is easy to take continuously.

[0019] In the present invention, the ratio of the weight of sesamins to the weight of ergothioneine or its salt in terms of ergothioneine (sesamins / ergothioneine) is preferably 0.1 or more, more preferably 0.15 or more, preferably 20 or less, more preferably 16 or less, and even more preferably 15 or less. In one embodiment, the ratio of the weight of sesamins to the weight of ergothioneine or its salt in terms of ergothioneine (sesamins / ergothioneine) is preferably 0.1 to 20, more preferably 0.15 to 16, and even more preferably 0.15 to 15. Compositions containing sesamins and ergothioneine or its salt in the above ratios can effectively suppress cellular senescence. The weight ratio of sesamins and ergothioneine or its salt in the composition is preferably as described above.

[0020] Cellular senescence refers to a state in which cell proliferation irreversibly ceases. While the factors contributing to individual aging are diverse, it has been found that in aging animals, the number of cells undergoing cellular senescence (senescent cells) increases with age, and that primary human cultured cells, when subcultured, eventually reach their mitotic limit and become senescent cells. Bioactive factors secreted by senescent cells are thought to adversely affect surrounding normal cells. Senescent cells also exhibit reduced cellular function. Suppressing cellular senescence is effective in preventing or improving conditions or diseases caused by cellular senescence or an increase in senescent cells. Examples of conditions or diseases caused by cellular senescence or an increase in senescent cells include decreased cell function, decreased organ function, decreased systemic function, cancer, and chronic inflammation. By suppressing cellular senescence and inhibiting the increase (accumulation) of senescent cells in the target population, it is expected that effects such as maintaining cell function, organ function, and systemic function can be obtained. Furthermore, it is expected that preventive or ameliorative effects on conditions or diseases such as cancer and chronic inflammation can be obtained.

[0021] The cellular senescence inhibitory effect of the present invention can be confirmed by known cellular senescence markers. Examples of cellular senescence markers include p16-INK4A (p16). p16 induces cellular senescence by arresting the cell cycle in the G1 phase. Therefore, increased expression of p16 promotes cellular senescence. In one embodiment, the composition of the present invention is preferably one that suppresses the expression of p16. In one embodiment, the composition of the present invention can be used to suppress the expression of p16.

[0022] As shown in the examples below, the use of sesamins and ergothioneine effectively suppressed the expression of p16, an indicator of cellular senescence. The p16 expression inhibitory effect of the combination of sesamins and ergothioneine was a synergistic effect superior to the effect (additive effect) predicted from the effects of using sesamins alone or ergothioneine or its salts alone. Sesamins enhanced the p16 expression inhibitory effect of ergothioneine or its salts. Therefore, a combination of one or more sesamins and ergothioneine or a salt thereof can be used as an active ingredient for suppressing cellular senescence. In one embodiment, the composition of the present invention can be used to suppress cellular senescence. Furthermore, as shown in the examples below, the combination of sesamins and ergothioneine showed an effect of extending the cell division lifespan in yeast. An extension of the cell division lifespan can be considered an extension of the individual's lifespan. Therefore, compositions containing sesamins and ergothioneine or their salts exhibit an effect of preventing or improving aging in individuals (anti-aging effect). In this specification, anti-aging refers to the prevention or improvement of aging in individuals.

[0023] Sesamins and ergothioneine or their salts are compounds found in natural products and foods and beverages, and have a history of being consumed. Therefore, from a safety standpoint, sesamins and ergothioneine or their salts are considered to pose little problem even if ingested daily, for example. According to the present invention, it is possible to provide a composition that contains highly safe substances as active ingredients and has the effect of suppressing cellular aging. The composition of the present invention may be a composition for suppressing cellular aging that contains one or more sesamins and ergothioneine or its salt as active ingredients.

[0024] In mammals such as humans, metabolism and other processes typically decline with age, leading to the assumption that the rate of removal of senescent cells slows down with age. Suppressing cellular senescence is considered particularly effective in inhibiting the increase of senescent cells, for example, in middle-aged and elderly individuals. Middle-aged and elderly individuals include the elderly.

[0025] By suppressing cellular senescence in subjects such as humans, the increase of senescent cells in the subject can be suppressed. In one embodiment, the composition of the present invention can be used to prevent or improve conditions or diseases such as decreased cellular function, decreased organ function, or decreased systemic function. In one embodiment, the composition of the present invention can be used to prevent or improve age-related decline in organ function, age-related decline in systemic function, etc., by suppressing cellular senescence. Age-related decline in organ function and age-related decline in systemic function are examples of symptoms of individual aging in living organisms. Furthermore, it has been reported that lifespan can be extended by suppressing the accumulation of senescent cells (Non-Patent Literature 2 above). The composition of the present invention is also useful for preventing or improving individual aging and can be used as an anti-aging composition. The individual is preferably a human. The composition of the present invention may be an anti-aging composition containing one or more types of sesamin and ergothioneine or a salt thereof as active ingredients. In one embodiment, the composition of the present invention can be suitably used as an anti-aging composition for preventing or improving aging by suppressing cellular senescence. The composition of the present invention may be an anti-aging composition that acts via the suppression of cellular senescence. By suppressing cellular senescence and preventing or improving the aging of an individual, a life-extending effect can also be expected. The composition of the present invention is useful, for example, for extending healthy life expectancy. In this specification, prevention includes preventing the onset of disease, delaying the onset of disease, reducing the incidence rate, and mitigating the risk of developing disease. Improvement of a condition or disease includes recovering from a condition or disease, alleviating the symptoms of a condition or disease, improving the symptoms of a condition or disease, slowing the progression of a condition or disease, and preventing it.

[0026] The compositions of the present invention can be applied to either therapeutic (medical) or non-therapeutic (non-medical) uses. Non-therapeutic means that the use does not involve medical procedures, i.e., human surgery, treatment, or diagnosis. The composition of the present invention can be in the form of food and beverages, cosmetics, pharmaceuticals, quasi-drugs, animal feed, etc. The composition of the present invention may be food and beverages, cosmetics, pharmaceuticals, quasi-drugs, animal feed, etc. used for inhibiting cellular aging, or it may be a material or formulation used in combination with these. The compositions of the present invention can be provided, for example, in the form of a formulation, but are not limited to this form. The formulation can also be provided as a composition as is, or as a composition containing the formulation. In one embodiment, a composition for inhibiting cellular senescence can also be called a cellular senescence inhibitor. An anti-aging composition can also be called an anti-aging agent.

[0027] The composition of the present invention may be either an oral composition or a parenteral composition. In one embodiment, the composition of the present invention is preferably an oral composition. Examples of oral compositions include food and beverages, oral pharmaceuticals, quasi-drugs, and animal feed, and are preferably food and beverages or oral pharmaceuticals, and more preferably food and beverages. In another embodiment, the composition of the present invention can be used as a topical skin preparation (topical skin composition). Topical skin preparations include cosmetics, pharmaceuticals, quasi-drugs, etc., and are preferably cosmetics.

[0028] The composition of the present invention may contain any additives and components in addition to one or more sesamins and ergothioneine or a salt thereof, as long as the effects of the present invention are not impaired. These additives and components can be selected according to the form of the composition, and those that can be used in food and beverages, cosmetics, pharmaceuticals, quasi-drugs, animal feed, etc., can be used. When the composition of the present invention is used in food and beverages, cosmetics, pharmaceuticals, quasi-drugs, animal feed, etc., the manufacturing method is not particularly limited and can be manufactured by general methods. In one embodiment, the composition of the present invention may consist of one or more sesamins and ergothioneine or a salt thereof.

[0029] For example, when the composition of the present invention is used as a food or beverage, various food and beverages can be made by blending one or more types of sesamin and ergothioneine or its salt with ingredients usable in food and beverages (e.g., food materials, food additives used as needed). Food and beverages are not particularly limited and include, for example, general food and beverages, health foods, health supplements, health drinks, foods with functional claims, foods for specified health uses, and food and beverages for sick people. The above-mentioned health foods, health supplements, foods with functional claims, foods for specified health uses, etc., can be used in various formulation forms such as fine granules, tablets, granules, powders, capsules, chewable tablets, dry syrups, syrups, liquids, beverages, drinks, and liquid foods.

[0030] When the composition of the present invention is used as a cosmetic, various cosmetic compositions can be made by blending one or more types of sesamin and ergothioneine or a salt thereof with ingredients that can be used in cosmetics (for example, carriers, additives, etc. that are acceptable in cosmetics). The product form of the cosmetic is not particularly limited, and examples include skincare cosmetics such as lotions, serums, masks, emulsions, creams, and sunscreens.

[0031] When the composition of the present invention is used as a pharmaceutical or quasi-drug, for example, one or more types of sesamin and ergothioneine or a salt thereof can be combined with a pharmacologically acceptable carrier, additives as needed, etc., to produce pharmaceuticals or quasi-drugs in various dosage forms. Such carriers, additives, etc., only need to be pharmacologically acceptable and usable in pharmaceuticals or quasi-drugs, and examples include one or more excipients, binders, disintegrants, lubricants, antioxidants, colorants, etc. The form of administration (ingestion) of the pharmaceutical or quasi-drug can be oral or parenteral (transdermal, transmucosal, enteral, injection, etc.). In one embodiment, when the composition of the present invention is used as a pharmaceutical or quasi-drug, it is preferable to use it as an oral pharmaceutical or quasi-drug. Dosage forms for oral administration include liquids, tablets, powders, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewables, etc. In another embodiment, the pharmaceutical or quasi-drug is preferably a topical skin preparation. The dosage form of the topical skin preparation is not particularly limited and can be any form, such as a solution, emulsion, cream, gel, powder, aerosol, mist, capsule, and sheet. Pharmaceuticals may also be pharmaceuticals for non-human animal use.

[0032] When the composition of the present invention is used as animal feed, one or more types of sesamin and ergothioneine or a salt thereof may be added to the feed. Feed additives may also be included in the feed. Examples of feed include livestock feed for cattle, pigs, chickens, sheep, horses, etc.; small animal feed for rabbits, rats, mice, etc.; and pet food for dogs, cats, small birds, etc.

[0033] The content of sesamin compounds in the composition of the present invention is not particularly limited and can be set according to its form and other factors. The total content of sesamin compounds in the composition of the present invention is preferably 0.001% by weight or more, more preferably 0.01% by weight or more, even more preferably 0.05% by weight or more, and preferably 10% by weight or less, and even more preferably 5% by weight or less. In one embodiment, the total content of sesamin compounds is preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, and even more preferably 0.05 to 5% by weight. The above total content is the sum of the content of sesamin compounds if two or more sesamin compounds are included. The content of sesamin compounds can be measured using known methods such as high-performance liquid chromatography (HPLC).

[0034] The amount of ergothioneine or its salt contained in the composition of the present invention is not particularly limited and can be set according to its form and other factors. The content of ergothioneine or its salt in the composition of the present invention is preferably 0.001% by weight or more, more preferably 0.01% by weight or more, even more preferably 0.05% by weight or more, and preferably 10% by weight or less, and even more preferably 5% by weight or less, in terms of ergothioneine. In one embodiment, the content of ergothioneine or its salt is preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, and even more preferably 0.05 to 5% by weight, in terms of ergothioneine, in the composition of the present invention. The content of ergothioneine or its salt can be measured using known methods such as HPLC. The amount of ergothioneine or its salt in terms of ergothioneine or similar expressions means the amount of ergothioneine when ergothioneine is used, and the value obtained by multiplying the number of moles of the salt by the molecular weight of ergothioneine when a salt of ergothioneine is used.

[0035] The compositions of the present invention can be ingested or administered by various methods known to the present invention, depending on their form or dosage form. In one embodiment, the compositions of the present invention are preferably ingested orally (administered orally). In another embodiment, when the compositions of the present invention are used as a topical skin preparation, they can be applied to the skin. The dosage (or intake) of the composition of the present invention is not particularly limited. The dosage of the composition of the present invention should be, for example, an amount that provides a cellular senescence inhibitory effect or an anti-aging effect, and should be set appropriately according to the form of administration, method of administration, the weight of the subject, etc.

[0036] In one embodiment, when the composition of the present invention is administered to a human (adult), the total dose of sesamin is preferably 0.5 mg or more, more preferably 1 mg or more, even more preferably 3 mg or more, and preferably 200 mg or less, more preferably 100 mg or less, and even more preferably 80 mg or less per day. In one embodiment, the total dose of sesamin is preferably 0.5 to 200 mg, more preferably 1 to 100 mg, and even more preferably 3 to 80 mg per day for a human (adult). In one embodiment, it is preferable to orally administer sesamin and / or episesamin to a human (adult) at a total dose of sesamin and episesamin of preferably 0.5 to 200 mg, more preferably 1 to 100 mg, and even more preferably 3 to 80 mg per day. The above total dose is the sum of the amounts of sesamin compounds used when two or more types of sesamin compounds are used. The above dose may be the dose per 60 kg of body weight.

[0037] In one embodiment, when the composition of the present invention is administered to a human (adult), the dose of ergothioneine or its salt is preferably 2 mg or more, more preferably 5 mg or more, preferably 50 mg or less, more preferably 25 mg or less, and still preferably 20 mg or less, in terms of ergothioneine per day. In one embodiment, when the composition of the present invention is administered to a human (adult), the dose of ergothioneine or its salt is preferably 2 to 50 mg, more preferably 2 to 25 mg, still preferably 5 to 25 mg, and particularly preferably 5 to 20 mg, in terms of ergothioneine per day. In one embodiment, it is preferable to orally administer the above-mentioned amount of sesamin and ergothioneine or its salt to a human being per day. It is also preferable to administer the above amount once or more per day, for example, once a day or in several divided doses (for example, 2 to 3 times). The above dosage may be the dosage per 60 kg of body weight. When using the composition of the present invention in humans for the purpose of obtaining a cellular senescence inhibitory effect or an anti-aging effect, it is preferable to use the above-mentioned range for the dosage of sesamins and ergothioneine or its salts. In one embodiment, the composition of the present invention can be used to administer to a human being the above-mentioned amount of sesamins and ergothioneine or a salt thereof per day.

[0038] The composition of the present invention preferably contains a total amount of sesamins of 0.5 to 200 mg per adult daily intake, more preferably 1 to 100 mg, and even more preferably 3 to 80 mg. The composition of the present invention preferably contains ergothioneine or a salt thereof in terms of ergothioneine equivalent of 2 to 50 mg per adult daily intake, more preferably 2 to 25 mg, even more preferably 5 to 25 mg, and particularly preferably 5 to 20 mg.

[0039] The composition of the present invention is preferably ingested or administered continuously. In one embodiment, the composition of the present invention is preferably ingested or administered continuously for at least one week, more preferably for at least four weeks, even more preferably for at least eight weeks, and particularly preferably for at least twelve weeks.

[0040] The subjects (or recipients) to whom the composition of the present invention is administered are not particularly limited. Preferably, they are humans or non-human mammals, and more preferably, humans. In one embodiment, potential recipients of the composition include individuals who require or desire the suppression of cellular senescence, or individuals who require or desire the prevention or improvement of aging symptoms. Examples of such individuals include middle-aged and elderly people. In one embodiment, the composition of the present invention is suitably used as a cellular senescence suppression composition or anti-aging composition for middle-aged and elderly people. Middle-aged and elderly people may be, for example, people aged 40 or older. In one embodiment, among middle-aged and elderly people, elderly individuals are preferred as the target. Elderly people may be, for example, people aged 60 or older or 65 or older. The composition of the present invention can also be used in healthy individuals for purposes such as the prevention of conditions or diseases that can be prevented or improved by the suppression of cellular senescence.

[0041] The types of cells targeted for cellular senescence suppression are not particularly limited, and examples include fibroblasts, epithelial cells, epidermal cells, basal cells, stem cells, adipocytes, nerve cells, muscle cells, skeletal muscle cells, smooth muscle cells, myofibroblasts, muscle satellite cells, satellite cells, osteoblasts, hepatocytes, hepatic stellate cells, renal cells, stromal cells, blood cells, immune system cells, endothelial cells, vascular endothelial cells, pericytes, lens cells, retinal cells, and the like. In one embodiment, the composition of the present invention is suitably used for suppressing cellular senescence of fibroblasts. Fibroblasts are essential cells for tissue support and structure formation in almost all organs.

[0042] The composition of the present invention may be marked with one or more functional indications selected from the group consisting of, for example, "inhibition of cellular senescence," "inhibition of senescent cell formation," "prevention of cellular senescence," "prevention of the formation of senescent cells," "inhibition of the increase of senescent cells," "inhibition of the accumulation of senescent cells," "delaying cellular senescence," "delaying the progression of cellular senescence," "inhibition of cellular senescence induction," "increase in the proportion of young cells," "extension of healthy lifespan," "healthy longevity," "young cells," "youthful overall appearance," and "youthful appearance." In one embodiment of the present invention, the composition of the present invention is preferably a food or beverage or a topical skin preparation bearing the above-mentioned label, more preferably a food or beverage or a cosmetic, and even more preferably a food or beverage. The above-mentioned label may also be an indication that it is used to obtain the above-mentioned function. The above-mentioned label may be affixed to the composition itself, or to the container or packaging of the composition.

[0043] The present invention also includes the following methods. A method for suppressing cellular senescence, comprising administering one or more types of sesamin and ergothioneine or a salt thereof. By administering one or more types of sesamins and ergothioneine or a salt thereof to a target, it is possible to effectively suppress cellular senescence in the target. Furthermore, an anti-aging effect can be obtained in the target. In the above method, the ratio of the weight of sesamins to the weight of ergothioneine or a salt thereof in terms of ergothioneine (sesamins / ergothioneine) is preferably 0.05 to 50.

[0044] The present invention also includes the following uses: Use of one or more types of sesamin and ergothioneine or its salt to suppress cellular senescence. In one embodiment, one or more types of sesamins and ergothioneine or a salt thereof can be suitably used to prevent or improve aging in an individual by suppressing cellular senescence. In the above use, the ratio of the weight of sesamins to the weight of ergothioneine or a salt thereof in terms of ergothioneine (sesamins / ergothioneine) is preferably 0.05 to 50. The above methods and uses may be therapeutic or non-therapeutic.

[0045] In the above method and use, one or more sesamins, ergothioneine or its salts, and preferred embodiments thereof are the same as in the composition of the present invention described above. The preferred range of the weight ratio of one or more sesamins and ergothioneine or its salts is also the same as in the composition of the present invention. In the above method and use, it is preferable to administer (ingest) one or more sesamins and ergothioneine or its salts to the target at least once a day, for example, once to several times a day (for example, two to three times). Oral administration (ingestion) is preferred as the method of administration. The above use is preferably in humans or non-human mammals, more preferably in humans.

[0046] In the above method and use, one or more types of sesamin and ergothioneine or a salt thereof in an amount (which can also be called an effective amount) that yields the desired effect may be used. The preferred dosage and target recipients of sesamin, ergothioneine or a salt thereof are the same as those of the composition of the present invention described above. It is preferable to administer sesamin and ergothioneine or a salt thereof using a composition containing them. Preferably, a composition containing one or more types of sesamin and ergothioneine or a salt thereof in the above-mentioned ratio (the composition of the present invention described above) is used. The present invention also includes a method for suppressing cellular senescence, which involves administering the compositions of the present invention described above. The use of the compositions of the present invention for suppressing cellular senescence is also included in the present invention.

[0047] In the above method and use, by preparing sesamins (or compositions containing them) and ergothioneine or its salt (or compositions containing it) separately, and administering them almost simultaneously, or administering one after the other while its effect is still present, the effects of one or more sesamins and ergothioneine or its salt (cellular senescence inhibitory effect or anti-aging effect) intended by the present invention can be obtained. Therefore, kits containing sesamins or compositions containing them, and ergothioneine or its salt, or compositions containing it, are also included in the scope of the present invention.

[0048] The present invention also includes the use of one or more sesamins and ergothioneine or a salt thereof for producing a composition for inhibiting cellular senescence or an anti-aging composition. The compositions for inhibiting cellular senescence, the anti-aging compositions, and preferred embodiments thereof are the same as those of the present invention described above. The present invention also includes the use of one or more sesamins and ergothioneine or a salt thereof for producing a composition for inhibiting cell senescence or an anti-aging composition, wherein the ratio of the weight of sesamins to the weight of ergothioneine or a salt thereof in the composition (sesamins / ergothioneine) is 0.05 to 50. In this specification, a numerical range expressed by a lower limit and an upper limit, i.e., "lower limit to upper limit," includes those lower and upper limits. For example, a range expressed as "1 to 2" means 1 or more and 2 or less, including 1 and 2. In this specification, the upper and lower limits may be any combination of ranges. Furthermore, all academic and patent documents cited herein are incorporated herein by reference. [Examples]

[0049] The present invention will be described more specifically below based on examples. However, the present invention is not limited to these examples.

[0050] The ergothioneine used in the experiment below was L-ergothioneine. For the sesamin-episesamin mixture, a mixture of sesamin and episesamin with a sesamin:episesamin (weight ratio) of 1:1 was used.

[0051] <Examples 1-2: Evaluation tests of the effects of sesamin / episesamin mixture and ergothioneine on cellular senescence> TIG-3 cells (human lung fibroblasts) were used, and the expression level of p16-INK4A (p16) in these cells was measured. The culture medium used in Examples 1-2, Reference Example 1, and Comparative Examples 1-4 was Eagle's Minimum Essential Medium supplemented with 10% fetal bovine serum. The H2O2 (200 μM) supplemented medium used for H2O2 treatment was prepared by adding H2O2 to the above medium. The H2O2 (200 μM) supplemented medium did not contain sesamins or ergothioneine.

[0052] In Examples 1 and 2, to investigate the effects on cellular senescence, cells were cultured in a medium containing sesamins and ergothioneine, and then treated with a medium supplemented with H2O2. Subsequently, the cells were cultured again in a medium containing sesamins and ergothioneine. In detail, a culture medium containing a mixture of sesamin and episesamin (SE) and ergothioneine at the concentrations listed in Table 1 below was used, with 4.0 × 10⁶ of the solution added. 4TIG-3 cells at a cell / well density were seeded in 6 wells and cultured for 24 hours at 37°C with 5% CO2. Table 1 shows the weight ratio of sesamins (sesamin-episesamin mixture (SE)) to ergothioneine in the culture medium (SE / ergothioneine). After 24 hours of culture, the medium containing the sesamin-episesamin mixture and ergothioneine was removed, and the cells in the wells were treated with H2O2 (200 μM) supplemented medium for 2 hours. Subsequently, the H2O2 (200 μM) supplemented medium was replaced with medium containing the sesamin-episesamin mixture and ergothioneine at the concentrations shown in Table 1, and cultured for 6 days. Cells cultured in 6 wells were harvested using 500 μL of QIAzol Lysis Reagent (QIAGEN). 100 μM chloroform (Nacalai Tesque Co., Ltd.) was added, and the upper 200 μL was collected. Then, 200 μL of isopropanol (Nacalai Tesque Co., Ltd.) was added and the mixture was centrifuged (12000 g, 10 min, 4°C). After removing the supernatant, 500 μL of 70% EtOH (Nacalai Tesque Co., Ltd.) was added to the pellet and centrifuged (7500 g, 5 min, 4°C). The supernatant was removed, the mixture was dissolved in 20 μL of DNase RNase-free water (Invitrogen), and measured using nanodrops. 10 μL of RNA sample was mixed with 10 μL of prepared RT master mix (Applied Biosystems, High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor), and reacted in a thermal cycler (Applied Biosystems). 20 μL of the synthesized cDNA was diluted 1 / 10 with DNase RNase-free water. A 15 μL prepared master mix (Takara, TB Green® Premix Ex Taq) was then used. TMP16 levels were measured using a Real-Time PCR system (Applied Biosystems, StepOnePlus-01) after mixing with II (Tli RNaseH Plus) and a 5 μL cDNA sample. The mean p16 expression level (n=6) was calculated. The results are shown in Figure 1 (Example 1) and Figure 2 (Example 2). Significant difference testing was performed using Student's t-test (vs H2O2 treatment) (*: p<0.05).

[0053] <Example 1: Evaluation test of cellular senescence in an untreated group> To investigate its effects on cellular senescence, a mixture of sesamin and episesamin, along with 1.0 × 10⁶ of ergothioneine-free culture medium, were used. 5 TIG-3 cells at a rate of cells / well were seeded in 6 wells and cultured for 24 hours at 37°C with CO2 (5%). After 24 hours of culture, the cells were cultured for a further 2 days without being treated with H2O2 (200 μM) supplemented medium for 2 hours, nor without being replaced with medium supplemented with a sesamin / episesamin mixture and ergothioneine. In other words, in Reference Example 1, TIG-3 cells were cultured for 24 hours in a medium without sesamins and ergothioneine, and then cultured for a further 2 days in the same medium. In Reference Example 1, treatment with H2O2 (200 μM) supplemented medium was not performed. Otherwise, p16 was measured in the same manner as in Example 1. The obtained results are shown in Figures 1 and 2.

[0054] <Comparative Example 1: Evaluation test of cellular senescence in the H2O2-treated group> To investigate its effects on cellular senescence, a mixture of sesamin and episesamin, along with 4.0 × 10⁶ of ergothioneine-free culture medium, were used. 4TIG-3 cells at a rate of cells / well were seeded in 6 wells and cultured for 24 hours at 37°C with CO2 (5%). After 24 hours of culture, the cells in the wells were treated with H2O2 (200 μM) supplemented medium for 2 hours. Subsequently, the cells were cultured for 6 days without changing to a medium supplemented with sesamin / episesamin mixture and ergothioneine. In other words, in Comparative Example 1, after the above treatment with H2O2 (200 μM) supplemented medium, the H2O2 (200 μM) supplemented medium was replaced with a medium that did not contain sesamin and ergothioneine, and the cells were cultured for another 6 days. The p16 was measured in the same manner as in Example 1. The obtained results are shown in Figures 1 and 2.

[0055] <Comparative Examples 2-4: Evaluation tests of cellular senescence using sesamin / episesamin mixture or ergothioneine> To investigate the effects on cellular senescence, a culture medium containing a mixture of sesamin and episesamin or ergothioneine at the concentrations listed in Table 1 below was used, with 4.0 × 10⁶ of cells added. 4 TIG-3 cells at a rate of cells / well were seeded in 6 wells and cultured for 24 hours at 37°C with 5% CO2. After 24 hours of culture, the medium supplemented with sesamin / episesamin mixture or ergothioneine was removed, and the cells in the wells were treated with H2O2 (200 μM) supplemented medium for 2 hours. Subsequently, the H2O2 (200 μM) supplemented medium was replaced with medium supplemented with sesamin / episesamin mixture or ergothioneine at the concentrations shown in Table 1, and the cells were cultured for 6 days. The rest of the procedure was the same as in Example 1, and p16 was measured. The results obtained are shown in Figure 1 (Comparative Examples 2 and 3) and Figure 2 (Comparative Examples 2 and 4). In Table 1, "H2O2 (μM)" refers to the H2O2 concentration in the culture medium supplemented with H2O2 (200 μM).

[0056] [Table 1]

[0057] <Result> Figures 1 and 2 are graphs showing the results of investigating the cellular senescence inhibitory effects of a sesamin / episesamin mixture and ergothioneine. The results (relative values) shown in Figures 1 and 2 are shown as relative values ​​of the p16 expression levels in the examples and comparative examples, with the p16 expression level in Reference Example 1 set to 1. Specifically, the relative value was obtained by dividing the p16 expression level in the examples (average value for n=6) by the p16 expression level in Reference Example 1 (average value for n=6) (p16 expression level in examples / p16 expression level in Reference Example 1). Similarly, for the comparative examples, the relative value was obtained by dividing the p16 expression level in the comparative examples (average value for n=6) by the p16 expression level in Reference Example 1 (average value for n=6). As shown in Figures 1 and 2, p16 increased approximately twice as much as the control group (Reference Example 1) with the 200 μM H2O2 treatment according to Comparative Example 1. Also, as shown in Figure 1, the treatments with the sesamin / episesamin mixture (1 μM) or ergothioneine (0.1 μM) according to Comparative Examples 2 and 3 did not result in any significant change in p16 expression compared to the H2O2 treatment group according to Comparative Example 1. On the other hand, treatment with the composition according to Example 1, which contains a sesamin / episesamin mixture (1 μM) and ergothioneine (0.1 μM) (with a weight ratio of sesamins to ergothioneine of 15.5), significantly suppressed p16 expression. Next, as shown in Figure 2, in the case of the sesamin / episesamin mixture (1 μM) according to Comparative Example 2, there was no significant change in the expression level of p16 compared to the H2O2-treated group according to Comparative Example 1, while in the case of the ergothioneine (10 μM) treatment according to Comparative Example 4, p16 expression was significantly suppressed. Treatment with the sesamin / episesamin mixture (1 μM) and the composition containing ergothioneine (10 μM) according to Example 2 (weight ratio of sesamins to ergothioneine: 0.155) significantly suppressed p16 expression, and the level of suppression was higher than that of the ergothioneine-only treatment (Comparative Example 4). From the results of the Examples and Comparative Examples, it was considered that the cellular senescence inhibitory effect of ergothioneine was enhanced by the sesamins. Based on the above, it was confirmed that the expression of p16 was synergistically suppressed by a composition containing a mixture of sesamin and episesamin and ergothioneine. In other words, it was confirmed that compositions containing sesamin and ergothioneine exhibit a synergistic anti-senescence effect due to the combination of both, and are effective even at low doses.

[0058] <Example 3: Confirmation test of life extension effect by sesamin / episesamin mixture and ergothioneine> The life-extending effects of sesamin / episesamin mixture and ergothioneine were investigated using yeast. Since the daughter yeast of yeast K6001 does not divide, the lifespan of the parent cell can be measured by counting the number of daughter cells that budding from the parent cell. In this study, yeast K6001 was cultured to the stationary phase in YPGalactose liquid medium (2% galactose, 1% yeast extract, 2% peptone). A portion of the yeast was collected and washed twice with water. The washed yeast was seeded on YPGlucose agar medium (2% galactose, 1% yeast extract, 2% peptone) containing sesamin / episesamin mixture (10 μM) and ergothioneine (10 μM). After culturing at 30°C for 2 days, the daughter cells were counted under a microscope. The weight ratio of sesamin compounds to ergothioneine in the medium (sesamin / episesamin mixture / ergothioneine) was 1.55. As a control, yeast was cultured in the same manner as above, except that a sesamin / episesamin mixture and ergothioneine were not added to the culture medium, and daughter cells were counted under a microscope.

[0059] <Result> Figure 3 is a graph showing the results of an investigation into the life-extending effects of sesamin / episesamin mixture and ergothioneine in yeast. The horizontal axis of the graph in Figure 3 shows the generation (number) of daughter cells that budded from a single parent cell. The dividing cells (%) on the vertical axis represent the percentage of parent cells that budded the daughter cells of the generation shown on the horizontal axis, relative to the total number of parent cells (100%) (Reference: Jarolim, et al. “A novel assay for replicative lifespan in Saccharomyces cerevisiae.” FEMS yeast research 5.2 (2004): 169-177). For example, if five daughter cells (generations 1-5) bud from a parent cell, that parent cell (dividing cell) is counted as generation 1-5, respectively. In Figure 3, black circles (●) represent the control, and white circles (○) represent yeast cultured in a medium containing sesamin / episesamin mixture (SE) and ergothioneine. As shown in Figure 3, compared to the control group, treatment with the sesamin / episesamin mixture and ergothioneine increased the number of daughter yeast cells and extended the lifespan of the yeast cells. Therefore, it was confirmed that compositions containing sesamin and ergothioneine exhibit a lifespan-extending effect due to the presence of both compounds.

Claims

1. It contains one or more types of sesamin, and ergothioneine or a salt thereof. A composition in which the ratio of the weight of sesamins to the weight of ergothioneine or its salt in terms of ergothioneine (sesamins / ergothioneine) is 0.05 to 50.

2. The composition according to claim 1, wherein the sesamins are sesamin and / or episesamin.

3. The composition according to claim 1 or 2, which is an oral composition.

4. The composition according to claim 1 or 2, which is a food or beverage.

5. The composition according to claim 1 or 2, which is a cosmetic.

6. A composition according to claim 1 or 2, used for suppressing cellular senescence.

7. The composition according to claim 1 or 2, which is an anti-aging composition.

8. The composition according to claim 1 or 2, which is marked with one or more expressions selected from the group consisting of "inhibits cellular aging," "inhibits the formation of senescent cells," "prevents cellular aging," "prevents the formation of senescent cells," "inhibits the increase of senescent cells," "suppresses the accumulation of senescent cells," "delays cellular aging," "delays the progression of cellular aging," "inhibits the induction of cellular aging," "increases the proportion of young cells," "extends healthy lifespan," "healthy longevity," "young cells," "youthful overall," and "youthful appearance."

9. A method for suppressing cellular senescence, comprising administering the composition described in claim 1 or 2.

10. Use of the composition according to claim 1 or 2 for suppressing cellular senescence.

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