Agent for removing senescent cells and agent for suppressing decrease in motor function

Abstract

The purpose of the present invention is to provide a preparation and a composition that are effective in removal of senescent cells and can be ingested even in the field of food and drink products. Provided is an agent for removing senescent cells, the agent containing an effective amount of at least one selected from the group consisting of Albizia julibrissin flowers, Albizia julibrissin bark, and extracts of these.

Description

Senescent cell remover and motor function inhibitor

[0001] The present invention relates to a senolytic agent. More specifically, the present invention relates to a senolytic agent that selectively eliminates senescent cells relative to normal cells.

[0002] Currently, developed countries are experiencing an aging society and average life expectancy is increasing, but the gap between this and healthy life expectancy is becoming a problem. Healthy life expectancy is a health indicator proposed by the World Health Organization (WHO) and refers to the period during which one can live an independent and healthy life both physically and mentally. The difference between healthy life expectancy and average life expectancy, which is the "unhealthy period" during which daily life is restricted, is said to be about 9 years for Japanese men and about 12 years for Japanese women.

[0003] As a countermeasure to the super-aging society, research and development into technologies for extending healthy lifespan is progressing, and attention is being paid to cellular senescence and the removal of senescent cells. Senescent cells are cells in which cell division has irreversibly stopped, and they exist not only in elderly people but also in young people and newborns.

[0004] It has become clear that cellular senescence is induced by DNA damage caused by stress such as ultraviolet rays and reactive oxygen species, as well as telomere shortening. It has also been suggested that inflammatory proteins secreted by senescent cells may cause various diseases, so there is a strong demand for technologies to control cellular senescence, both to suppress the induction of senescent cells and to remove the senescent cells that have already formed.

[0005] For example, Patent Document 1 reports that sodium-glucose cotransporter 2 (SGLT2) inhibitors have the effect of eliminating senescent cells. Specifically, the SGLT2 inhibitor canagliflozin has been confirmed to eliminate senescent cells in obese model mice, arteriosclerosis model mice, progeria model mice, and the like.

[0006] JP 2022-101541 A JP 2010-235548 A

[0007] However, the removal of senescent cells is itself difficult, and few techniques have been reported that can selectively remove senescent cells. Furthermore, there have been no reports of plant-based ingredients that can be ingested in foods and beverages and are effective in removing senescent cells.

[0008] Therefore, an object of the present invention is to provide a formulation and composition that can be ingested in the food and beverage sector and is effective in removing senescent cells.

[0009] In order to solve the above problems, the present inventors conducted extensive research and discovered that Albizia julibrissin, a plant commonly consumed in the food industry, has the effect of removing senescent cells, leading to the completion of the present invention. Furthermore, the present inventors also discovered that Albizia julibrissin has the effect of suppressing the decline of motor function.

[0010] That is, the present invention provides the formulations and compositions listed below.

[0011] Item 1. A senolytic agent containing an effective amount of at least one selected from the group consisting of rhododendron bark, rhododendron flower, and extracts thereof.

[0012] Item 2. The senolytic agent according to Item 1, which is labeled with at least one or more indications selected from the group consisting of "reducing senescent cells," "removing senescent cells," "reducing the proportion of senescent cells," "promoting the reduction of senescent cells," and "suppressing decline in motor function."

[0013] Item 3. The senolytic agent according to Item 1 or 2, wherein the senescent cell removal is performed by selectively removing senescent cells relative to normal cells.

[0014] Item 4. The senolytic agent according to any one of Items 1 to 3, wherein the ratio (A / B) of (A) the 50% lethal concentration (LC50) in normal cells to (B) the 50% lethal concentration in senescent cells is greater than 1.1.

[0015] Item 5. The senolytic agent according to any one of Items 1 to 4, which is a food product, cosmetic product, or pharmaceutical product.

[0016] Item 6. An agent for suppressing motor function decline, comprising an effective amount of at least one selected from the group consisting of rhododendron bark, rhododendron flower, and extracts thereof.

[0017] Item 7. A food composition for reducing senescent cells, removing senescent cells, decreasing the proportion of senescent cells, promoting the reduction of senescent cells, or suppressing a decline in motor function, comprising an effective amount of at least one selected from the group consisting of unicorn flower, unicorn bark, and extracts thereof.

[0018] Item 8. Use of at least one selected from the group consisting of euonymus flower, euonymus bark, and extracts thereof, for producing a composition for reducing senescent cells, removing senescent cells, decreasing the proportion of senescent cells, promoting the reduction of senescent cells, or suppressing a decline in motor function.

[0019] Item 9. The use of at least one selected from the group consisting of unicorn flower, unicorn bark, and extracts thereof according to Item 8, wherein the composition is a food, cosmetic, or pharmaceutical composition.

[0020] It should be noted that Patent Document 2 does not disclose or suggest the ability of Nemu to remove senescent cells.

[0021] According to the present invention, by using Albizia julibrissin, which can be ingested in the food and beverage sector, it is possible to effectively remove senescent cells. Furthermore, by using Albizia julibrissin, it is possible to suppress the decline in motor function.

[0022] Figure 1 is a graph showing the results of the senescent cell removal ability in Test Example 1. Figure 2 is a graph showing the results of the motor function confirmation test for the control group and the Nelumbo nucifera flower extract group (0.1 mg / mL) in Test Example 2. Figure 3 is a graph showing the results of the motor function confirmation test for the control group and the Nelumbo nucifera bark extract group (0.01 mg / mL) in Test Example 2.

[0023] [Senolytic Agent] In one embodiment, the senolytic agent of the present invention contains Albizia julibrissin.

[0024] (Albizia julibrissin) Albizia is a deciduous tree of the Fabaceae family, subfamily Mimosaideae. Albizia is also called Mimosa, and its name comes from the sleep behavior it performs at night, where its leaflets close and hang down.

[0025] The Japanese dogwood is found in Honshu, Shikoku, Kyushu, and other parts of Japan, and also grows wild in Iran, Afghanistan, China, the Korean Peninsula, and other areas, so it can be easily obtained from these areas.

[0026] In addition, the flowers and skin of the jasmine are sun-dried and used as a medicinal herb, and they are also brewed and drunk to relieve stress.

[0027] The part of the Synthetic Musk to be used is not particularly limited, and may be the whole plant, or may be leaves, fruits, flowers, branches, trunks, roots, seeds, or a mixture of at least two or more of them. In one embodiment, the part of the Synthetic Musk to be used preferably includes flowers and / or bark, and more preferably includes flowers and bark.

[0028] The rhododendron may be used fresh or dried (for example, freeze-dried). When extracting from the rhododendron, it may be processed into a crushed or powdered state before use, for example, from the viewpoint of improving extraction efficiency.

[0029] The compound may be, for example, crushed or pulverized in the form of chips, powder, granules, or paste, and among these, it is preferable to use powder.

[0030] The method for processing the plant into a crushed or pulverized material can be any known method and is not particularly limited. For example, the plant may be crushed or pulverized by any method commonly used by those skilled in the art using equipment or tools such as a crusher, mill, blender, mixer, or stone mill.

[0031] Before crushing or pulverizing, the plant body may be dried. The drying method may be any known method, and is not particularly limited, but examples thereof include freeze-drying, drying under reduced pressure, air drying, and heat drying.

[0032] When using an extract of the rhododendron bark, the method is not particularly limited and can be any conventional extraction method used for extracting plant materials. Examples of extraction methods include, but are not limited to, solvent extraction, steam distillation, compression (direct, high temperature, or low temperature), and supercritical extraction. A combination of these extraction methods may also be used, such as crushing followed by solvent extraction.

[0033] The extraction solvent is not particularly limited as long as it is a solvent commonly used for extracting plant materials. The extraction solvent may be, for example, an organic solvent, an aqueous solvent, or a mixture of an organic solvent and an aqueous solvent, but is preferably an aqueous solvent or a mixture of an organic solvent and an aqueous solvent, and more preferably an aqueous solvent.

[0034] Examples of organic solvents include alcohol, acetone, benzene, esters, ethyl acetate, hexane, chloroform, and diethyl ether, with alcohol being preferred. The alcohol is not particularly limited as long as it is a typical solvent used in the extraction of plant materials, and examples include monohydric alcohols having 1 to 5 carbon atoms, such as ethanol, methanol, propyl alcohol, isopropyl alcohol, and butyl alcohol, and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin.

[0035] The aqueous solvent is not particularly limited as long as it contains water, and examples thereof include water, physiological saline, and buffer solutions. Examples of buffer solutions include phosphate buffer solutions, sodium phosphate buffer solutions, sodium carbonate buffer solutions, citrate buffer solutions, citrate phosphate buffer solutions, acetate buffer solutions, and Tris buffer solutions. Preferred aqueous solvents are water and / or physiological saline solutions. The pH of the aqueous solvent is not particularly limited.

[0036] Of these extraction solvents, from the viewpoint of significantly exhibiting the effects of the present invention, at least one selected from the group consisting of water, ethanol, 1,3-butylene glycol, and propylene glycol is preferred, water and / or ethanol is more preferred, and water or aqueous ethanol is even more preferred.

[0037] When a mixture of an organic solvent and an aqueous solvent is used as the extraction solvent, the content of the aqueous solvent in the total amount of the extraction solvent is not particularly limited, and examples thereof include 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, and 95% by mass or more.

[0038] The extraction temperature is not particularly limited, but examples include -50°C to 150°C, 0°C to 100°C, 5°C to 95°C, 10°C to 95°C, 15°C to 95°C, 30°C to 90°C, 50°C to 90°C, 70°C to 90°C, and 80°C to 90°C.

[0039] From the viewpoint of improving extraction efficiency, it is preferable to carry out the extraction while stirring or shaking, for example. The extraction time can be appropriately determined depending on the part to be used, such as leaves, fruits, flowers, branches, stems, roots, seeds, etc. The extraction time can also be appropriately determined depending on the state of the rhododendron, i.e., whether it is fresh or dried, or whether it is crushed or pulverized, or the processed state thereof. The extraction time is usually 1 minute to 72 hours, preferably 10 minutes to 48 hours, and most preferably 30 minutes to 2 hours.

[0040] For example, the extract of Synthetic Bean may be an extract obtained by immersing Synthetic Bean flowers and / or bark, or crushed Synthetic Bean flowers and / or bark, in water and / or an organic solvent and filtering the residue; a solution obtained by removing the solvent from this extract; a fine powder of these; or a solution obtained by dissolving, dispersing, or diluting the extract or the solvent-removed material using an appropriate solvent; and commercially available products may also be used. As an example of another extraction method, Synthetic Bean extract may be extracted after processing Synthetic Bean flowers and / or bark, such as by steaming. Furthermore, the leaves and branches associated with the Synthetic Bean flowers and / or bark may be removed, or they may be used as is.

[0041] The content of the Synthetic Rice plant or an extract thereof is adjusted appropriately depending on the type and amount of other ingredients, dosage form, etc., and is not limited, and may be, for example, 0.01% by mass or more, 0.05% by mass or more, 0.1% by mass or more, 0.5% by mass or more, 1% by mass or more, 10% by mass or more, 20% by mass or more, 30% by mass or more, 40% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, etc. Furthermore, the content of the Synthetic Rice plant or an extract thereof may be, for example, 100% by mass or less, and examples thereof include 90% by mass or less, 80% by mass or less, 70% by mass or less, 60% by mass or less, 50% by mass or less, 40% by mass or less, 30% by mass or less, 20% by mass or less, 10% by mass or less, etc. The content of the Nemu plant or its extract relative to the total amount of the composition may be, for example, 0.01 to 100 mass%, 0.1 to 100 mass%, 1 to 100 mass%, 5 to 100 mass%, 10 to 100 mass%, 20 to 100 mass%, 30 to 100 mass%, 40 to 100 mass%, 50 to 100 mass%, 0.01 to 90 mass%, 0.1 to 90 mass%, 1 to 90 mass%, 5 to 90 mass%, %, 10-90% by mass, 20-90% by mass, 30-90% by mass, 40-90% by mass, 50-90% by mass, 0.01-80% by mass, 0.1-80% by mass, 1-80% by mass, 5-80% by mass %, 10-80% by mass, 20-80% by mass, 30-80% by mass, 40-80% by mass, 50-80% by mass, 0.01-70% by mass, 0.1-70% by mass, 1-70% by mass, 5-70% by mass %, 10-70% by mass, 20-70% by mass, 30-70% by mass, 40-70% by mass, 50-70% by mass, 0.01-60% by mass, 0.1-60% by mass, 1-60% by mass, 5-60% by mass %, 10-60% by mass, 20-60% by mass, 30-60% by mass, 40-60% by mass, 0.01-50% by mass, 0.1-50% by mass, 1-50% by mass, 5-50% by mass, 10-50% by mass %, 20-50 mass%, 30-50 mass%, 0.01-40 mass%, 0.1-40 mass%, 1-40 mass%, 5-40 mass%, 10-40 mass%, 20-40 mass%, 0.01-30 Examples include mass%, 0.1 to 30 mass%, 1 to 30 mass%, 5 to 30 mass%, 10 to 30 mass%, 0.01 to 20 mass%, 0.1 to 20 mass%, 1 to 20 mass%, 5 to 20 mass%, etc.

[0042] [Uses] The senescent cell-removing agent of the present invention effectively removes senescent cells, and is preferably suitable for selectively removing senescent cells over normal cells. As used herein, "senescent cells" refer to cells that have lost the ability to proliferate through cell division. The transformation into senescent cells is sometimes described as a defense mechanism that stops cell division to prevent cancer when DNA is irreparably damaged by stress, such as ultraviolet light. However, if senescent cells remain in the body without being eliminated, they are known to secrete inflammatory proteins and cause chronic inflammation. Furthermore, as used herein, "normal cells," which can be contrasted with senescent cells, refer to cells that have not lost the ability to proliferate through cell division.

[0043] Senescent cells have been identified using markers such as increased p16 gene and β-galactosidase activity, and it is becoming clear that removing senescent cells from aging individuals can suppress age-related changes. For example, in nonalcoholic steatohepatitis (NASH), senescent cells accumulate and inflammation may occur, but it has been reported that removing senescent cells can improve inflammation.

[0044] Even if there are anti-inflammatory ingredients for symptomatic treatment of inflammation caused by the accumulation of senescent cells, simply suppressing inflammation cannot be a fundamental solution as long as senescent cells accumulate. Furthermore, while various ingredients have been screened for anti-aging purposes, there have not been enough reports of ingredients that can suppress the generation of senescent cells. Furthermore, there are currently few reports of technologies that can distinguish senescent cells from normal cells once they have been generated and accumulated and selectively remove only the senescent cells.

[0045] The accumulation of senescent cells does not necessarily occur only in the elderly or sick, but can also occur in young people and newborns. Therefore, rather than relying on pharmaceuticals, there is a need for technology that can selectively remove senescent cells using ingredients that can be ingested on a daily basis. However, there have been few research reports from this perspective.

[0046] In the examples (test examples) described below, etoposide, an inducer of cellular senescence, was applied to human liver cancer-derived HepG2 cells to obtain senescent cells, and the effects of at least one selected from the group consisting of euonymus flower, euonymus bark, and extracts thereof, along with quercetin and other substances known to have the ability to remove senescent cells, were then confirmed.

[0047] As a result, it was confirmed that, compared to quercetin, at least one selected from the group consisting of euonymus flower, euonymus bark, and extracts thereof, has a significantly greater ability to selectively remove senescent cells.

[0048] Therefore, at least one selected from the group consisting of unicorn flower, unicorn bark, and extracts thereof can be suitably used as a selective agent for removing senescent cells, which can effectively remove senescent cells, preferably selectively remove senescent cells over normal cells.

[0049] The ability to selectively remove senescent cells relative to normal cells (senescent cell removal ability) can be calculated by determining the ratio (A / B) of (A) the 50% lethal concentration in normal cells to (B) the 50% lethal concentration in senescent cells, as described in the Examples below. The senescent cell removal ability is not limited as long as the effects of the present invention are achieved, but for example, the ratio (A / B) is preferably greater than 1.1, and may be 1.2 or more, 1.5 or more, 2 or more, 2.5 or more, 3 or more, 3.5 or more, 4 or more, 4.5 or more, 5 or more, 5.5 or more, 6 or more, 7 or more, 8 or more, 9 or more, etc.

[0050] [Agent for suppressing motor dysfunction] In one embodiment, the agent for suppressing motor dysfunction of the present invention contains GABA. The contents described in the embodiment of the senolytic agent for removing senolytic cells also apply to this agent for suppressing motor dysfunction.

[0051] A decline in motor function is caused by various factors, such as lack of exercise and disease, and also progresses gradually with aging. A decline in motor function can lead to conditions or symptoms such as sarcopenia, frailty, and locomotive syndrome. Furthermore, a decline in muscle strength and balance can increase the risk of falls and fractures. Furthermore, a decline in motor function can lead to a lack of exercise, which can lead to the onset or worsening of lifestyle-related diseases.

[0052] In the examples (test examples) described below, a control group without any additives was compared with a group to which unicorn extract was added, and the effect of at least one selected from the group consisting of unicorn flowers, unicorn bark, and their extracts in suppressing motor function decline was confirmed.

[0053] The formulations and compositions of the present invention can be added to or mixed with foods and beverages, cosmetics, pharmaceuticals, quasi-drugs, pet foods, and feeds. Alternatively, they can be used as they are in pharmaceuticals, quasi-drugs, foods and beverages, pet foods, and feeds. Furthermore, the formulations and compositions of the present invention can be used as foods and beverages that explicitly or implicitly claim to have functions such as senescent cell removal and suppression of motor function decline, i.e., health foods, functional food products, foods for patients, and foods for specified health uses. Even if the above functionality is not explicitly or implicitly claimed, they can be used as so-called doctor's supplements recommended or offered by doctors in internal medicine, orthopedic surgery, veterinary clinics, and the like in hospitals and / or clinics. When the above functionality is explicitly claimed, at least one of the following may be labeled, but is not limited to, "reducing senescent cells," "removing senescent cells," "reducing the proportion of senescent cells," "promoting the reduction of senescent cells," and "suppressing motor function decline." In addition, the effects attributed to the above functionality include "improvement of blood liver function enzyme levels," "maintenance of cognitive function," "maintenance of muscle strength," "maintenance of walking ability," "maintenance of muscle mass, muscle strength and muscle endurance necessary for living an independent daily life," "maintenance of agility," "reduction of fasting blood glucose levels," "moderating the rise in blood glucose levels after meals," "reduction of blood triglycerides," "maintenance of vascular flexibility which declines with age," "maintenance of kidney filtering function," "reduction of temporary swelling of the face and legs," "reduction in frequency of urination," "suppression of dizziness when standing," "maintenance of bone health," At least one or more claims selected from the group consisting of "maintaining bone density," "reducing knee discomfort," "supporting flexibility and mobility of knee joints," "improving skin moisture retention," "maintaining skin elasticity," "regrowing and promoting hair growth," "regulating the stomach," "improving visual function," "reducing transient physical and mental fatigue," "reducing biological age," "improving sarcopenia," "improving frailty," "improving locomotive syndrome," "improving lifestyle-related diseases," and "maintaining balance function" may be made, but are not limited to these.

[0054] When the above functionality is not explicitly stated, it is also possible to use the formulations and compositions of the present invention with a label such as, for example, "for those concerned about senescent cells, those who wish to remove senescent cells, those who wish to suppress or reduce senescent cells, those who wish to reduce the proportion of senescent cells, those concerned about their liver function or kidney function values, those who feel muscle weakness, those concerned about blood sugar levels, those concerned about bone density, those who feel discomfort in their knee joints, those concerned about their skin condition, those concerned about their stomach condition, those concerned about their vision, those who feel fatigue, those concerned about walking speed, or similar labeling.

[0055] The dosage form of the formulation and composition of the present invention is not particularly limited, and includes oral and parenteral formulations, with oral formulations being preferred. Examples of the oral formulation include solid or powder formulations such as fine granules, granules, tablets, capsules, and pills, as well as liquid formulations such as suspensions, emulsions, syrups, and extracts. Examples of parenteral formulations include injections, topical preparations, and poultices.

[0056] The formulations and compositions of the present invention may consist of at least one selected from the group consisting of unicorn flowers, unicorn bark, and extracts thereof, or may contain at least one selected from the group consisting of unicorn flowers, unicorn bark, and extracts thereof.

[0057] When the formulations and compositions of the present invention are oral dosage forms, they may contain other additives, such as excipients, binders, disintegrants, emulsifiers, lubricants, flow enhancers, diluents, preservatives, colorants, flavorings, corrigents, stabilizers, humectants, antiseptics, antioxidants, or suspending agents. Examples of additives include, but are not limited to, gelatin, sodium alginate, starch, corn starch, sucrose, lactose, glucose, mannitol, carboxymethylcellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soybean lecithin, sucrose, fatty acid esters, talc, magnesium stearate, polyethylene glycol, magnesium silicate, anhydrous silicic acid, and synthetic aluminum silicate.

[0058] The formulations and compositions of the present invention can be administered to humans, but may also be administered to animals other than humans, including pets such as dogs, cats, horses, ponies, donkeys, cows, pigs, sheep, goats, rabbits, monkeys, hamsters, guinea pigs, mice, rats, ferrets, squirrels, and birds; livestock such as cows, chickens, and pigs; and animals kept on ranches, zoos, etc. (horses, sheep, goats, monkeys, elephants, lions, cheetahs, etc.).

[0059] The composition of the present invention may be a food or drink, a cosmetic, a pharmaceutical, a quasi-drug, a pet food, a feed, or the like.

[0060] The food and drink products include food and beverages. Examples of food and beverages include beverages (soft drinks (coffee, cocoa, juice, mineral drinks, tea drinks (green tea, black tea, oolong tea), almond drinks, sports drinks, energy drinks, nutritional drinks, etc.), milk drinks, lactic acid bacteria drinks, yogurt drinks, carbonated drinks, alcoholic beverages, spreads (custard cream, butter cream, peanut cream, chocolate cream, cheese cream, etc.), pastes (fruit paste, vegetable paste, sesame paste, seaweed paste, etc.), Western confectionery (chocolate, donuts, pies, muffins, waffles, gum, Gummy candies, jellies, candies, cookies, crackers, biscuits, snacks, cakes, puddings, etc.), Japanese sweets (candy, rice crackers, karinto, arare, dumplings, ohagi, daifuku, mamemochi, mochi, bean paste, manju, castella, anmitsu, yokan, etc.), frozen desserts (ice cream, popsicles, sorbet, etc.), retort foods (curry, stew, beef bowl, Chinese rice bowl, rice porridge, porridge, miso soup, soup, meat sauce, demi-glace sauce, meatballs, hamburger steak, oden, red rice, yakitori, chawanmushi, etc.), instant foods (instant ramen, instant vermicelli soup, instant soup hot water) Leaf noodles, instant udon noodles, instant soba noodles, instant yakisoba noodles, instant spaghetti, instant wonton noodles, instant shiruko (sweet red bean paste), miso soup base, powdered soup base, powdered juice base, pancake mix, etc.), bottled and canned goods, jelly foods (jelly, agar, jelly drinks, etc.), seasonings (soy sauce, mirin, vinegar, miso, dressings, umami seasonings, compound seasonings, sauces, mayonnaise, ketchup, furikake (rice toppings), tempura sauce, noodle sauce, dashi stock, Chinese soup base, bouillon, yakiniku sauce, cold shabu-shabu sauce, curry roux, stew roux, etc.), dairy products (milk, cheese, yogurt, butter, fresh cream) etc.), processed fruits (jam, marmalade, fruits in syrup, dried fruits etc.), processed grain foods (noodles, pasta, bread, rice vermicelli etc.), pickles (takuan, narazuke, kimchi pickles, fukujinzuke, pickled radishes, pickled Chinese cabbage, pickled mustard, pickled eggplant, lightly pickled vegetables, pickles etc.), pickle bases (instant pickle bases, kimchi pickle bases etc.), fish products (kamaboko, chikuwa, hanpen etc.), livestock products (ham, sausage, salami, bacon etc.), delicacies (skimmed dried squid, skimmed cod, salted sea urchin, salted squid, salted octopus, mirinboshi filefish, mirinboshi pufferfish, smoked squid, salted konowata etc.),Examples of such foods include dried foods (seasoned seaweed, etc.), prepared foods (simmered dishes, deep-fried dishes, stir-fried dishes, grilled dishes, simmered dishes, vinegared dishes, etc.), frozen foods (fried shrimp, croquettes, spring rolls, pork cutlets, shumai, gyoza, takoyaki, meat buns, bean-jam buns, etc.), baby foods (e.g., rice and noodle dishes, fish dishes, meat dishes, egg dishes, dairy dishes, vegetable dishes, soups, fruits, juices, hydration drinks, tea, desserts, sweets, sauces, and furikake seasonings); powdered milk (e.g., general infant formula, special infant formula, and powdered milk for pregnant and nursing women); enteral nutritional foods (e.g., enteral nutritional supplements, concentrated liquid diets, nutritional supplements, nutritional supplements, enteral nutritional supplements, flavors for concentrated liquid diets, and desserts); and other foods (e.g., nutritional supplements, blender meals, thickened foods, foods for examinations, cereal foods, foods for sports, and emergency foods).

[0061] These foods or beverages may contain food additives and food ingredients, such as, but not limited to, antioxidants, flavorings, acidulants, colorings, emulsifiers, preservatives, seasonings, sweeteners, spices, pH adjusters, stabilizers, vegetable oils, animal oils, sugars and sugar alcohols, vitamins, organic acids, fruit juice extracts, vegetable extracts, grains, beans, vegetables, meat, and seafood, either alone or in combination of two or more.

[0062] Tablets can be prepared by mixing a powdered active ingredient with a pharmaceutically acceptable carrier (such as an excipient) and compressing the mixture, and confectionery tablets such as candy can be prepared by pouring the mixture into a mold. Tablets can also be sugar-coated to form sugar-coated tablets. Furthermore, tablets can be single-layer tablets or layered tablets such as double-layer tablets.

[0063] Powdered granules such as granules may be prepared by various granulation methods (extrusion granulation, pulverization granulation, dry compaction granulation, fluidized bed granulation, tumbling granulation, high-speed stirring granulation, etc.), and tablets can be prepared by an appropriate combination of the above-mentioned granulation methods and tableting methods (wet tableting, direct tableting), etc.

[0064] Capsules can be prepared by filling a capsule (soft or hard capsule) with powder (powder, granules, etc.) by a conventional method.

[0065] A liquid preparation can be prepared by dissolving or dispersing each component in an aqueous medium (purified water, purified water containing ethanol, etc.) which is a carrier component, filtering or sterilizing the resulting mixture if necessary, filling the resulting mixture into a predetermined container, and sterilizing the resulting mixture. The preferred dosage form of a solid preparation is a capsule or tablet, and soft capsules (soft capsules) are more preferred.

[0066] Soft capsules are preferred by users because they have a smooth surface and are easy to swallow. Common methods for manufacturing soft capsules include the flat plate method, the rotary method, and the seamless method.

[0067] In the rotary method (punching method), a sheet-like capsule shell sandwiches the flowing filling material and forms a capsule shape along the holes of a rotating cylindrical mold. On the other hand, in the seamless method (dropping method), the capsule shell composition and the filling material are simultaneously discharged from multiple concentric nozzles to form a seamless capsule shape.

[0068] The base material for the soft capsule shell is not particularly limited, but may be starch, pullulan, cellulose, polyvinyl alcohol, gelatin, succinated gelatin, etc., with starch, gelatin, and succinated gelatin being preferred, and gelatin and succinated gelatin being more preferred. These may be used alone or in combination of two or more.

[0069] The daily oral intake or administration amount of unicorn flower, unicorn bark, or an extract thereof for an adult can be determined appropriately based on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, and other factors. The daily oral intake or administration amount of unicorn flower, unicorn bark, or an extract thereof for an adult is, for example, preferably 0.1 mg or more, more preferably 1 mg or more, even more preferably 10 mg or more, particularly preferably 20 mg or more, and most preferably 30 mg or more. It may also be 100 mg or more, 200 mg or more. The daily oral intake or administration amount of unicorn flower, unicorn bark, or an extract thereof for an adult is, for example, preferably 2000 mg or less, more preferably 1500 mg or less, even more preferably 1000 mg or less, and particularly preferably 500 mg or less. The daily oral intake or administration amount of the rhododendron flower, rhododendron bark, or an extract thereof for an adult is, for example, preferably 0.1 to 2000 mg, more preferably 1 to 1500 mg, even more preferably 10 to 1000 mg, even more preferably 20 to 500 mg, and even more preferably 30 to 500 mg.

[0070] The oral intake or dosage per day for an adult may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form.

[0071] The composition of the present invention can be ingested or administered once to several times a day, typically 1 to 6 times a day, 1 to 3 times a day, 1 to 2 times a day, or at any period and interval, but once a day is preferred.

[0072] Next, the present invention will be specifically explained by way of examples, but the present invention is not limited to the following examples.

[0073] [Preparation of Test Formulations] (Example 1. Ethanol Extract 1) 3.0 g of dried powder of the flower of the genus Aster was added to 30 mL of 99.5% ethanol and stirred at room temperature for 30 minutes to perform extraction. After that, the residue was removed by a conventional method, and the mixture was dried to obtain 0.1326 g of a dried product.

[0074] (Example 2 Ethanol Extract 2) 3.0 g of dried powder of Synthetic Root Bark was added to 30 mL of 99.5% ethanol and stirred at room temperature for 30 minutes to perform extraction. After that, the residue was removed by a conventional method, and the mixture was dried to obtain 0.1551 g of a dried product.

[0075] (Example 3. Hot Water Extract 1) 27 mL of water was added to 3.0 g of dried powder of the flower of the same name, and the mixture was extracted by stirring for 1 hour at 90° C. After that, the residue was removed by a conventional method, and the mixture was dried to obtain 0.7323 g of a dried product.

[0076] (Example 4. Hot Water Extract 2) 27 mL of water was added to 3.0 g of dried powder of Synthetic Chestnut Bark, and the mixture was stirred for 1 hour at 90° C. to carry out extraction. After that, the residue was removed by a conventional method, and the mixture was dried to obtain 0.1851 g of a dried product.

[0077] Example 5: Alkaline Extract: 25 mL of water was added to 3.0 g of dried powder of the rhododendron amurense, and the mixture was adjusted to pH 9.0 (±0.1) using 0.5 mol / L sodium hydroxide solution, followed by adding water to a total of 33 g. The mixture was stirred at 70°C for 1 hour to extract. After adjusting the pH to 7.0 using 0.5 mol / L hydrochloric acid, the residue was removed by conventional methods and the mixture was dried to obtain 0.5802 g of dried product.

[0078] Test Example 1: Senescent Cell Elimination Ability Verification Test (1) Preparation of Senescent Cells Human liver cancer-derived HepG2 cells were seeded at 10,000 cells per well onto a 12-well plate and treated with 10 μM etoposide (Sigma-Aldrich) (containing 0.1% DMSO) for two days to induce cellular senescence. To confirm the induction of cellular senescence, the cells were treated with 1 μM SPiDER-βGal (Dojindo) and observed for fluorescent signal using a GFP filter (excitation wavelength: 470 / 40 nm, absorption wavelength: 525 / 50 nm) on an all-in-one fluorescence microscope BZ-9000 (Keyence). Conversion of SPiDER-βGal to a fluorescent substance caused by β-galactosidase activity in senescent cells was observed in etoposide-treated cells.

[0079] (2) Evaluation of senescent cell elimination activity The ratio of the 50% lethal concentration (LC50) in normal (non-senescent) cells to the LC50 in senescent cells was determined. LC50 is the concentration required to kill half of the cells in a plate.

[0080] HepG2 cells (8,000 cells) were seeded in a 96-well plate and treated with 10 μM etoposide (containing 0.1% DMSO) for two days to induce cellular senescence. Subsequently, the cells were treated with the test formulations prepared above, with 0.179-40 μg / mL of Symphytum officinalis flower and Symphytum officinalis bark extract added to RPMI medium (HEPES-free) containing 0.5% fetal bovine serum (ThermoFisher). After two days, cell counts were measured using CellTiter-Glo (Promega) and EnVision (PerkinElmer) based on luminescence intensity. LC50 values ​​were calculated using GraphPad Prism 9 (GraphPad Software) based on the measured values ​​of the treatment concentration. The results of the senescent cell removal ability calculated based on the following formula are shown in Figure 1.

[0081] How to calculate senescent cell elimination capacity: Senescent cell elimination capacity = LC50 in normal (non-senescent) cells / LC50 in senescent cells

[0082] As shown in Figure 1, when the senescent cell removal ability, which is the ability to induce cell death specific to senescent cells, was calculated using the above formula, the values ​​obtained for the Unicorn flower extract were 9.8 (hot water extract), 5.0 (ethanol extract), and 4.7 (alkali extract), while the Unicorn bark extract gave values ​​of 4.2 (hot water extract) and 3.9 (ethanol extract). When the senescent cell removal ability of quercetin, which is known to have an effect of removing senescent cells, was examined under the same conditions, the result was 1.1, confirming that the Unicorn flower extract and Unicorn bark extract have a higher senescent cell removal ability than the known quercetin.

[0083] [Test Example 2. Motor Function Verification Test] (1) Rearing of Drosophila Wild-type Drosophila (W １１１８The male fruit flies (strain) were used. After emergence, they were reared under an environment of 25±3°C temperature, 70±20% humidity, and a 12-hour light / dark cycle. The control group was fed a regular diet (corn flour 4.4 w / w%, dry yeast Y4 5 w / w%, powdered agar 0.44 w / w%, glucose 5.6 w / w%, propionic acid 0.44 w / w%, bokinin 0.41 w / w%). The euonymus flower extract group (0.1 mg / mL) was fed a diet supplemented with euonymus flower extract (prepared in the same manner as in Example 3. Hot Water Extract 1) at a final concentration of 0.1 mg / mL. The euonymus bark extract group (0.01 mg / mL) was fed a diet supplemented with euonymus flower extract (prepared in the same manner as in Example 4. Food containing a final concentration of 0.01 mg / mL of Synechococcus bark extract, prepared in the same manner as for hot water extract 2, was placed at the bottom of a vial (diameter 22 mm, height 96 mm) and allowed to be eaten ad libitum. The food was transferred to a vial containing new food once a week.

[0084] (2) Evaluation of Motor Function The motor function of Drosophila was evaluated by performing a climbing assay to assess negative gravitaxis. The climbing assay was performed under the following conditions. Ten Drosophila per vial were transferred to an empty vial containing no food. The vial was then tapped to drop the flies to the bottom, and the flies' climbing up the wall of the vial was videotaped. Ten seconds after the flies were dropped to the bottom, the number of flies in the upper third, middle third, or lower third of the vial was counted. Each flies was assigned a score of 3 points for the top of the vial, 2 points for the center, and 1 point for the bottom. The total score for each group was calculated, and the average climbing score was calculated by dividing the total score by the number of flies. The average climbing scores for each age are shown in Tables 1 and 2 and Figures 2 and 3, respectively.

[0085] (3) Evaluation of the effect of Nembu flower extract on improving motor function

[0086]

[0087] As shown in Table 1 and Figure 2, the average climbing score, which indicates the motor ability of Drosophila, was calculated and the group administered Nembu flower extract achieved a value of 1.94 at 36 days of age. The control group had a score of 1.49 at the same age, confirming that Nembu flower extract has the ability to suppress motor function decline.

[0088] (4) Evaluation of the effect of Neomum bark extract on motor function

[0089]

[0090] As shown in Table 2 and Figure 3, the average climbing score, which indicates the motor ability of Drosophila, was calculated and the value was 2.04 at 36 days of age in the group administered with Nemu skin extract. Since the score in the control group at the same age was 1.23, it was confirmed that Nemu skin extract has the ability to suppress the decline in motor function.

Claims

1. An aging cell remover comprising at least one selected from the group consisting of Albizia julibrissin Durazz., cortex albiziae, and their extracts in an effective amount.

2. The aging cell remover according to claim 1, labeled with at least one selected from the group consisting of "reducing aged cells", "removing aging cells", "decreasing the proportion of aging cells", "promoting the decrease of aging cells", and "suppressing the decline of motor function".

3. The aging cell remover according to claim 1, wherein the removal of aging cells selectively removes aging cells from normal cells.

4. The aging cell remover according to claim 1, characterized in that the ratio (A / B) of (A) the 50% lethal concentration (LC50) in normal cells to (B) the 50% lethal concentration in aging cells is more than 1.

1.

5. The aging cell remover according to claim 1, which is a food, a cosmetic, or a pharmaceutical.

6. A motor function decline inhibitor comprising at least one selected from the group consisting of Albizia julibrissin Durazz., cortex albiziae, and their extracts in an effective amount.

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