Healthy life expectancy extension composition

Abstract

The present disclosure addresses the problem of providing a composition that contains 5-hydroxyequol and that can be used in a new application. The present disclosure also addresses another problem of providing a method for extending healthy life expectancy, the method comprising bringing 5-hydroxyequol into contact with a subject.　The present disclosure provides a composition for extending healthy life expectancy, the composition containing 5-hydroxyequol. Also provided is a method for producing a composition containing 5-hydroxyequol, the method comprising producing 5-hydroxyequol from genistein through fermentation. The present disclosure also provides a method for extending healthy life expectancy, the method comprising bringing 5-hydroxyequol into contact with a subject.

Description

Composition for extending healthy lifespan

[0001] The present disclosure relates to a composition for extending healthy lifespan, which contains 5-hydroxy equol. The present disclosure also relates to a method for extending healthy lifespan, which includes administering 5-hydroxy equol to a subject.

[0002] Genistin, an isoflavone abundantly contained in leguminous plants such as soybeans and kudzu, is converted into genistein by the action of digestive enzymes or enzymes (such as β-glucosidase) produced by intestinal bacteria when it enters the body of humans or animals. Genistein is converted into 5-hydroxy equol by the action of some intestinal bacteria (Non-Patent Document 1).

[0003] 5-Hydroxy equol has estrogen-like activity and has also been reported to have 3-β-hydroxysteroid dehydrogenase inhibitory activity, and thus may be available for the prevention or treatment of aldosterone and glucocorticoid excess (Patent Document 1). Furthermore, 5-hydroxy equol has also been reported to have an effect of extending the average lifespan (Non-Patent Document 2). In addition, 5-hydroxy equol can be safely and easily ingested, for example, as a component contained in a food composition. Thus, 5-hydroxy equol is known to have useful attributes, and it is considered that it can be effectively utilized, for example, as a component of a composition having functionality for the health of organisms.

[0004] If unknown attributes of 5-hydroxy equol can be discovered and it can be found that 5-hydroxy equol is suitable for use in new applications, 5-hydroxy equol can be utilized more effectively.

[0005] Japanese Patent Application Laid-Open No. 2003-81875

[0006] Matties et al., Appl. Environ. Microbiol., Vol. 74, No. 15, pp. 4847-4852 (2008); Zhang et al., J. Chin. Pharm. Sci., Vol. 23, No. 6, pp. 378-384 (2014)

[0007] This disclosure aims to provide a composition containing 5-hydroxyequol that can be used for novel applications. Another objective is to provide a method for extending healthy life expectancy, which includes contacting 5-hydroxyequol with a target.

[0008] The inventors have surprisingly discovered that 5-hydroxyequol has the effect of extending healthy life expectancy, and have completed the present invention. This disclosure includes the following embodiments: [1] A composition for extending healthy life expectancy, comprising 5-hydroxyequol.

[0009] The present disclosure can provide compositions containing 5-hydroxyequol for use in novel applications. According to one aspect of the present disclosure, a composition containing 5-hydroxyequol for extending healthy lifespan can be provided. According to another aspect of the present disclosure, a method for extending healthy lifespan can be provided, which includes contacting a subject with 5-hydroxyequol.

[0010] Figure 1 shows a schematic of the administration method of 5-hydroxyequol to nematodes. Figure 2 shows the C-HAS population analysis data and the qHaS calculated from it in nematode groups administered with 0.1% DMSO (A), 10 μM 5-hydroxyequol (B), 30 μM 5-hydroxyequol (C), and 50 μM 5-hydroxyequol (D).

[0011] The following describes in detail one embodiment of this disclosure, but the scope of this disclosure is not limited to the embodiment described herein, and various modifications can be made without departing from the spirit of this disclosure. Each configuration and combination thereof in each embodiment is an example, and additions, omissions, substitutions, and other modifications can be made as appropriate without departing from the spirit of this disclosure. This disclosure is not limited by embodiments. Each aspect disclosed herein can be combined with any other features disclosed herein. Furthermore, if multiple upper and lower limits are described for a particular parameter, any upper and lower limits can be combined to form a suitable numerical range. Also, the lower and / or upper limits of the numerical range described herein may be replaced with numerical values ​​within that numerical range, as shown in the examples. The expression "X to Y" indicating a numerical range means "X or more and Y or less". If a particular description described for one embodiment also applies to other embodiments, that description may be omitted in the other embodiments.

[0012] <Composition> In one embodiment, a composition comprising 5-hydroxyequol for extending healthy life expectancy is disclosed. In today's aging society, while the extension of average life expectancy is remarkable, the gap between healthy life expectancy and average life expectancy is still not small. The composition disclosed herein can effectively extend healthy life expectancy, thereby improving quality of life (QOL).

[0013] [5-Hydroxyequol] 5-Hydroxyequol can be prepared by methods well known to those skilled in the art. For example, 5-Hydroxyequol may be 5-Hydroxyequol prepared by chemical synthesis, or 5-Hydroxyequol produced from genistein by fermentation. The microorganisms that carry out fermentation are not particularly limited, as long as they are microorganisms capable of producing 5-Hydroxyequol from genistein. The microorganisms are preferably bacteria, and more preferably intestinal bacteria. Furthermore, the microorganisms are preferably anaerobic microorganisms. The anaerobic microorganisms may be obligate anaerobic microorganisms or facultative anaerobic microorganisms. Non-limiting examples of microorganisms include those belonging to the genera Asaccharobacter, Adlercreutzia, Coriobacteriaceae, and Slackia isoflavoniconvertens. 5-hydroxyequol produced from genistein by fermentation can be prepared, for example, according to the method described in Japanese Patent Publication No. 2018-198546. Preferably, 5-hydroxyequol is 5-hydroxyequol produced from genistein by fermentation. This 5-hydroxyequol tends to effectively extend healthy life expectancy.

[0014] [Healthy Life Expectancy] Healthy life expectancy, when referring to the healthy life expectancy of an individual as described below, is the period during which a person can live without their daily life being restricted by health problems. Alternatively, when referring to the healthy life expectancy of a tissue or cell as described below, healthy life expectancy may be the period during which the tissue or cell can survive while maintaining a healthy state.

[0015] Healthy life expectancy can be evaluated by known methods. Known methods include C-HAS (C-HAS+ Co., Ltd.), an automated measurement technology for healthy life expectancy using nematodes, which will be described later. In this case, the effect of the sample on extending healthy life expectancy can be effectively evaluated by quadrant Healthable score (qHaS), which will be described later.

[0016] Healthy lifespan is not particularly limited, as long as it is the healthy lifespan of an organism. An organism may be an individual, tissue, or cell, preferably an individual. An organism may also be an animal or a plant, preferably an animal. That is, an individual is an individual of an animal or plant, preferably an individual of an animal. A tissue or cell is a tissue or cell derived from an animal or plant, preferably a tissue or cell derived from an animal.

[0017] The animals are not limited as long as they are multicellular eukaryotes belonging to the animal kingdom. The animals may be vertebrates or invertebrates, but are preferably vertebrates. Vertebrates are preferably mammals, birds, reptiles, amphibians, or fish, and are more preferably mammals. Non-limiting examples of mammals include humans, monkeys, dogs, cats, horses, cattle, pigs, sheep, goats, rabbits, guinea pigs, hamsters, mice, rats, etc. Non-limiting examples of invertebrates include nematodes (e.g., C. elegans) or fruit flies (e.g., D. melanogaster). The animals are preferably humans.

[0018] The cells may be cells contained in an individual or tissue, or cells isolated from an individual or tissue. Cells isolated from an individual or tissue may be primary cultured cells, subcultured cells, or cell lines. The origin of the cells is not limited; for example, they may be cells derived from the endoderm, ectoderm, and / or mesoderm, or cells derived from epithelial cells, endocrine cells, muscle cells, nerve cells, and / or blood cells (including lymphocytes, granulocytes, red blood cells, etc.). The cells may be, for example, embryonic stem cells, induced pluripotent stem cells, adult stem cells, differentiated cells, blood cells, hematopoietic cells, epithelial cells, exocrine cells, endocrine cells, connective tissue cells, adipocytes, osteocytes, smooth muscle cells, striated muscle cells, nerve cells, sensory cells, cardiac cells, hepatocytes, gastric cells, intestinal cells, lung cells, kidney cells, and / or germ cells.

[0019] The tissue, if of animal origin, may be, for example, epithelial tissue, connective tissue, muscle tissue, and / or nerve tissue. Furthermore, the tissue is not limited by the organ from which it originates.

[0020] Healthy life expectancy may be evaluated using health indicators. If the health indicators of an individual, tissue, and cell are higher than the reference value, it can be determined that the individual, tissue, and cell are in a healthy state, and the period during which this state is maintained can be defined as healthy life expectancy. Health indicators may include, for example, an early death index, a disability index, a comprehensive early death / disability index, a QOL index, etc. Health indicators may also include, for example, cell activity indicators (including cells contained in individuals and tissues). Non-limiting examples of cell activity indicators include mitochondrial activity and autophagy levels. Reference values ​​can be set appropriately by those skilled in the art, but may include, for example, the minimum value, mean value, median value, etc., of the health indicators that enable individuals, tissues, and cells to perform the desired activities.

[0021] [Extending Healthy Lifespan] "Extending healthy lifespan" means, for example, that when the composition disclosed herein is administered to a subject, healthy lifespan is extended compared to when it is not administered (or to a negative control (control individual or control group)). Furthermore, "extending healthy lifespan" may mean that when the composition disclosed herein is administered to a subject, healthy lifespan is extended by a statistically significant margin compared to a negative control, or that healthy lifespan is extended by, for example, 5% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or less, 70% or more, 80% or more, 90% or more, 100% or more, 200% or more, 300% or more, 400% or more, or 500% or more.

[0022] Whether 5-hydroxyequol has an effect on extending healthy lifespan in living organisms (including cells, tissues, and individuals) can be effectively evaluated using C-HAS (C-HAS+ Inc.), an automated measurement technology for healthy lifespan using nematodes. In C-HAS, the nematode C. elegans (wild strain N2) is used as a model organism, and still images of nematodes to which the sample has been administered are acquired over time using the live cell imaging analysis instrument IncuCyte® S3 (Essen bio science Inc.). By superimposing the acquired sequential images and observing the movement (phase difference) of the nematodes from the degree of overlap, active nematodes (active and healthy nematodes), immobile nematodes (unhealthy nematodes that remain in place but only their heads and tails move), and dead nematodes (nematodes that are not moving at all) can be distinguished. Using this distinction as an indicator, we can detect the period from the beginning of adulthood until immobility, and the period from immobility until death. Based on these periods, the period from birth until immobility is defined as the healthy period, and the period from immobility until death is defined as the unhealthy period. Here, the healthy period corresponds to healthy lifespan, and the sum of the healthy period and the unhealthy period corresponds to lifespan (or average lifespan). By defining individuals with a longer healthy lifespan than the average healthy lifespan of the control group as healthy and individuals with a shorter healthy lifespan as unhealthy, and by defining individuals with a longer lifespan than the average lifespan of the control group as long-lived and individuals with a shorter lifespan as short-lived, we can classify nematodes into the following four groups: 1. Healthy, short-lived group ("short-lived" group) 2. Unhealthy, long-lived group ("standard" group) 3. Healthy, long-lived group ("healthy long-lived" group) 4. Unhealthy, short-lived groups ("unhealthy" groups) Furthermore, based on the above classification, the percentage of each group can be determined as the percentage of the number of individuals in each group relative to the total number of individuals in groups 1 to 4. From the percentage of each group, quadrant Healthable score (qHaS) can be calculated based on the following formula.qHaS = (3. Percentage of healthy, long-lived group) - [(4. Percentage of unhealthy, short-lived group) + (1. Percentage of healthy, short-lived group)] The effect of 5-hydroxyequol on extending healthy life expectancy can be effectively evaluated based on qHaS.

[0023] The qHaS of the composition of the present disclosure is preferably 5 or more, more preferably 10 or more, and particularly preferably 15 or more, or preferably 5 to 50, more preferably 10 to 50, and particularly preferably 15 to 50.

[0024] The compositions of this disclosure can extend the healthy lifespan of nematodes by preferably 1 day or more, more preferably 2 days or more, even more preferably 5 days or more, or preferably 1 to 20 days, more preferably 2 to 15 days, and even more preferably 5 to 15 days, in the evaluation of healthy lifespan using C-HAS. The compositions of this disclosure can extend the average lifespan of nematodes by preferably 1 day or more, more preferably 2 days or more, even more preferably 5 days or more, or preferably 1 to 20 days, in the evaluation of healthy lifespan using C-HAS.

[0025] The compositions of this disclosure can extend the healthy lifespan of a human being, preferably by more than one year, more preferably by more than two years, even more preferably by more than five years, or preferably by 1 to 20 years, more preferably by 2 to 15 years, and even more preferably by 5 to 15 years, when administered to a human being.

[0026] [Decline of physical and mental functions associated with aging] "Extending healthy life expectancy" may include mitigating the decline of physical and mental functions associated with aging. In this case, "extending healthy life expectancy" may mean extending the period during which the decline of physical and mental functions associated with aging is mitigated. Physical and mental functions may include motor functions. Therefore, a composition for extending healthy life expectancy may be a composition for mitigating the decline of physical and mental functions associated with aging, and may also be a composition for mitigating the decline of motor functions associated with aging.

[0027] Mitigating the decline in physical and mental functions of an individual with aging may include reducing or supporting the likelihood of symptoms and / or diseases occurring in an individual that increase in frequency or tend to increase with age, improving or supporting the health of patients who are experiencing such symptoms and / or diseases, and / or reducing or supporting the likelihood of patients experiencing such symptoms and / or diseases becoming more unhealthy.

[0028] Symptoms and / or diseases that tend to increase in frequency with age are not particularly limited within the scope in which the effects of the present invention can be obtained.

[0029] For example, symptoms and / or diseases that tend to increase in frequency with age may include frailty, locomotive syndrome (also called locomo), and / or sarcopenia. Frailty is a state in which various organ function changes and a decline in reserve capacity occur with age, and vulnerability to external stress is increased. Sarcopenia is a state characterized by a progressive and systemic decrease in muscle mass and strength, and is accompanied by physical disability, a decline in quality of life, and an increased risk of death. Locomotive syndrome is a state in which daily life is restricted due to musculoskeletal disorders, and care and assistance are required or at increased risk. Sarcopenia is considered a concept encompassed within locomotive syndrome, and sarcopenia and locomotive syndrome are considered a concept encompassed within frailty. Frailty, locomotive syndrome, and / or sarcopenia are generally accompanied by phenomena such as decreased myosin expression, increased mitochondrial fragmentation, and decreased physical activity. These phenomena can be effectively analyzed in nematodes, which serve as a model organism. Therefore, the effect of the compositions disclosed herein on mitigating the decline of physical and mental functions in individuals associated with aging can be effectively evaluated in an experimental system using nematodes as a model (for example, the experimental system using C-HAS (C-HAS+ Inc.) described above).

[0030] Furthermore, symptoms and / or diseases that tend to increase in frequency with age may include decreased muscle strength, increased disease risk, skin aging (increased depth and / or number of wrinkles, increased age spots, warts, sagging, etc.), hair aging (graying, decreased volume, deterioration of hair quality), obesity (including visceral and subcutaneous obesity), and / or decreased digestive function (including decreased chewing function, oral function, esophageal function, gastrointestinal function, etc.). Non-limiting examples of disease risk include the risk of developing cancer, muscle diseases, neurological diseases, etc. Non-specific examples of cancer include carcinoma, lymphoma, blastoma, sarcoma, leukemia, or lymphoid malignancy, while more specific examples include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung cancer including lung adenocarcinoma and squamous cell carcinoma of the lung, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatocellular carcinoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine carcinoma, salivary gland carcinoma, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver carcinoma, anal carcinoma, penile carcinoma, and head and neck cancer. Non-specific examples of muscle diseases include Parkinson's disease, amyotrophic lateral sclerosis, myasthenia gravis, muscular dystrophy, and polymyositis. Non-specific examples of neurological disorders include Parkinson's disease, dementia, Guillain-Barré syndrome, Pick's disease, and Huntington's disease. Non-specific examples of dementia include Alzheimer's disease.

[0031] [Prevention of Sudden Death in Youth] Extending healthy life expectancy may include preventing sudden death in youth. In other words, a composition for extending healthy life expectancy may also be a composition for preventing sudden death in youth. Sudden death is death without signs of disease or other illness, and refers to the sudden death of a healthy individual without going through a period of ill health (for example, within 24 hours of onset). Note that sudden death does not include death due to external causes (for example, death due to a traffic accident). Sudden death in youth not only shortens healthy life expectancy, but is also difficult to prevent because it occurs without signs of disease or other illness. However, it is believed that the composition disclosed herein can effectively prevent sudden death in youth.

[0032] The prevention of sudden death in young people can be evaluated using the C-HAS (C-HAS+ Inc.) described above to assess healthy life expectancy by reducing the percentage of individuals in "1. Healthy, short-lived group" relative to the total number of nematodes (total number of individuals in "1. Healthy, short-lived group", "2. Unhealthy, long-lived group", "3. Healthy, long-lived group", and "4. Unhealthy, short-lived group"). The composition of this disclosure can reduce the percentage of the healthy, short-lived group in the total number of nematodes by preferably 5 points or more, more preferably 10 points or more, even more preferably 20 points or more, or by 5 to 50 points, more preferably 10 to 40 points, and even more preferably 20 to 30 points, in the evaluation of healthy life expectancy using C-HAS.

[0033] [Dosage Form] The compositions of this disclosure can be administered to a subject by any method within the scope of obtaining the effects of the present invention. For example, if the subject is an individual, the compositions of this disclosure can preferably be administered to the individual orally. If the subject is a tissue or cells, the tissue or cells administered with the compositions of this disclosure may be transplanted into an individual. In this case, the compositions of this disclosure may be used not only for the purpose of extending the healthy lifespan of the tissue or cells, but also for the purpose of extending the healthy lifespan of the individual.

[0034] [Additives] The compositions of this disclosure may contain any additives to the extent that the effects of the present invention are obtained. Non-limiting examples of additives include excipients, binders, fillers, disintegrants, surfactants, lubricants, dispersants, buffers, diluents, preservatives, flavoring agents, fragrances, and the like.

[0035] [Food Compositions] The compositions of this disclosure may be food compositions. Food compositions may be compositions that are food in themselves, or compositions that are used as ingredients for food. Food compositions may be general foods, health functional foods (including nutritional functional foods, foods for specified health uses, and foods with functional claims), health foods, functional foods, nutritional supplements, foods for the sick, or food additives. Food compositions may take any form within the scope of obtaining the effects of the invention, for example, in the form of beverages, powders, pastes, jellies, capsules, tablets, etc. Food compositions may be taken alone or mixed with other foods and beverages. Other non-exclusive examples of food and beverages include: oil-based products such as edible oils, dressings, mayonnaise, and margarine; liquid foods such as soups, dairy drinks, soft drinks, tea drinks, alcoholic beverages, energy drinks, jelly drinks, and functional beverages; carbohydrate-containing foods such as rice, noodles, and bread; processed meat products such as ham and sausages; processed seafood products such as fish cakes, dried fish, and salted seafood; processed vegetable products such as pickles; semi-solid foods such as jelly and yogurt; fermented foods such as miso and fermented beverages; various confectionery products such as Western-style sweets, Japanese-style sweets, candies, gums, gummies, frozen desserts, and ice cream; retort products such as curry, thickened sauces, and Chinese soups; and instant foods such as instant soups and instant miso soup, as well as microwaveable foods.

[0036] The food composition can be manufactured as one or more food products. The food composition can be administered to an individual at any frequency or interval, within the scope of achieving the effects of the invention.

[0037] A food composition, or a food made from the same, may be a food composition or food that displays a function related to mitigating the decline in physical and mental functions of an individual associated with aging, as described above. For example, if a food composition, or a food made from the same, is a functional food, the functional food may be a food that displays a statement such as "contains 5-hydroxyequol," "contains microbial metabolites of daidzein," and / or "contains microbial metabolites of genistein." Furthermore, if a food composition, or a food made from the same, is a food for specified health uses or a food with functional claims, the food for specified health uses or the food with functional claims may be a food that displays the following functions for 5-hydroxyequol, for example. - Supports the maintenance of muscle mass and strength necessary for maintaining the physical strength required for independent daily living. - Helps maintain or suppress the decline of muscle mass and strength. - Helps improve walking ability. - Supports the bending and straightening of the knee joint (smoothness of movement). - Increases or retains skin moisture. - Suppresses the decline in skin moisture (hydration) and elasticity. - Alleviates dry skin. - Improves digestive health. - Helps reduce visceral fat. - Helps improve BMI. - Suitable for overweight individuals. - Maintains cognitive function (attention, perception, or recall of recognized things). - Maintains memory.

[0038] In one embodiment, the food composition contains 5-hydroxyequol in an amount of preferably 0.01 to 20% by weight, more preferably 0.1 to 10% by weight, and even more preferably 1 to 5% by weight, based on the total weight of the composition.

[0039] [Pharmaceutical Compositions] The compositions of this disclosure may be pharmaceutical compositions. A pharmaceutical composition may be a composition that is a pharmaceutical in itself, or a composition that is used as a material for a pharmaceutical. A pharmaceutical composition may be a prescription drug, an over-the-counter drug, or a quasi-drug.

[0040] The pharmaceutical composition can be formulated into one or more dosage forms. The pharmaceutical composition can be administered to an individual systemically or locally at any dosing frequency and / or interval within the range where the effects of the present invention can be obtained. Non-limiting examples of administration routes include oral administration, intravenous administration, intramuscular administration, intrathecal administration, sublingual administration, rectal administration, ophthalmic administration, otic administration, transmastoid administration, and transdermal administration.

[0041] The pharmaceutical composition can take any form within the range where the effects of the present invention can be obtained. For example, the pharmaceutical composition may take forms such as tablets, granules, powders, capsules, emulsions, suspensions, syrups, injections or drip agents (sterile solutions, suspension agents, etc.).

[0042] The pharmaceutical composition, or a medicament using this as a material, may be used to alleviate the decline in the physical and mental functions of an individual associated with aging as described above. In this case, alleviating the decline in the physical and mental functions of an individual associated with aging may be treating and / or preventing the above-described symptoms and / or diseases.

[0043] "Treatment" includes the reduction, alleviation, and mitigation of symptoms and / or disease symptoms, and "prevention" includes the defense against and suppression of the progression of future onset of symptoms and / or diseases. Desirable treatment effects by treatment include remission of symptoms, improvement of direct or indirect pathological results of the disease, reduction of the rate of progression of symptom exacerbation, recovery or alleviation of the disease state, and improvement of prognosis. Treatment and / or prevention includes administering a pharmaceutical composition or a medicament for the purpose of such treatment and / or prevention to a subject diagnosed with symptoms and / or diseases.

[0044] In one embodiment, the pharmaceutical composition contains 5-hydroxy equol, preferably 0.01 to 20% by weight, more preferably 0.1 to 10% by weight, still more preferably 1 to 5% by weight, based on the total weight of the composition.

[0045] [Cell Culture Reagents] The compositions of this disclosure may be added to culture media as reagents for culturing cells and / or tissues. In this case, the compositions of this disclosure can be suitably used, for example, for the purpose of maintaining or preserving cells and / or tissues in culture media in a healthy state for a long period of time. The compositions of this disclosure can also be part of a special cell freezing solution in cell therapy applications using primary cells that require cryopreservation.

[0046] [Cell Exposure] The compositions of the present disclosure may be compositions in which cells are exposed to 5 to 100 μM, preferably 10 to 50 μM, of 5-hydroxyequol. Alternatively, the compositions of the present disclosure may be compositions comprising 5-hydroxyequol in a dose that exposes cells to 5 to 100 μM, preferably 10 to 50 μM, of 5-hydroxyequol. These compositions tend to be able to more effectively extend healthy lifespan. The cells may be the cells described above, for example, cells contained in an individual or tissue, or cells isolated from an individual or tissue. If the cells are cells contained in an individual or tissue, 5-hydroxyequol may take the form of a 5 to 100 μM, preferably 10 to 50 μM, 5-hydroxyequol solution within the individual or tissue after administration to the individual or tissue, or after the individual has ingested 5-hydroxyequol. In this case, the 5-hydroxyequol solution may use a body fluid such as blood as the solvent. Therefore, exposure of cells to 5-hydroxyequol in a concentration of 5-100 μM, preferably 10-50 μM, may occur, for example, after an individual has been administered 5-hydroxyequol or a composition containing it, or after an individual has ingested 5-hydroxyequol or a composition containing it, and the cells within the individual's body are exposed to a bodily fluid (e.g., blood) containing 5-hydroxyequol in a concentration of 5-100 μM, preferably 10-50 μM.

[0047] <Method for Producing Composition> In one embodiment, a method for producing the above-described composition is disclosed, which includes producing 5-hydroxy equol from genistein by fermentation. Fermentation is not particularly limited as long as it is a method for producing a compound using microorganisms. The microorganisms are not particularly limited as long as they have the ability to produce 5-hydroxy equol from genistein. The microorganisms are preferably bacteria, more preferably intestinal bacteria. Also, the microorganisms are preferably anaerobic microorganisms. The anaerobic microorganisms may be obligate anaerobic microorganisms or facultative anaerobic microorganisms. Non-limiting examples of the microorganisms include microorganisms belonging to the genus Asaccharobacter and / or microorganisms belonging to the genus Adlercreutzia. Producing 5-hydroxy equol from genistein by fermentation can be carried out, for example, according to the method described in JP-A-2018-198546. The composition of the present disclosure can be effectively produced by this production method, and can be efficiently produced, for example, on an industrial scale. Also, the composition produced by this production method tends to have a high effect of extending the healthy life span.

[0048] <Method for Extending Healthy Life Span> In one embodiment, a method for extending the healthy life span is disclosed, which includes bringing 5-hydroxy equol into contact with a subject. This method can effectively extend the healthy life span, thereby improving the quality of life (QOL).

[0049] Bringing 5-hydroxy equol into contact with a subject can be preferably carried out by administering the above-described composition to the subject, or, when the subject is an individual, by the individual ingesting the above-described composition. The subject and the method of administration are as described above.

[0050] A method for extending healthy lifespan may involve exposing cells to a 5-hydroxyequol solution of 5-100 μM, preferably 10-50 μM. The cells may be the aforementioned cells, for example, cells contained in an individual or tissue, or cells isolated from an individual or tissue. When 5-hydroxyequol is administered to an individual or tissue, or when an individual ingests 5-hydroxyequol, 5-hydroxyequol may take the form of a 5-hydroxyequol solution of 5-100 μM, preferably 10-50 μM, within the individual or tissue. In this case, the 5-hydroxyequol solution may use a body fluid such as blood as the solvent. Therefore, contact with a subject may involve, for example, administering 5-hydroxyequol to a subject (for example, an individual that is a human or a non-human animal) (for example, by oral administration), and then exposing cells (for example, cells within an individual) to a body fluid (for example, blood) containing 5-hydroxyequol in a 5-100 μM, preferably 10-50 μM solution. Exposure of cells to a 5-hydroxyequol solution at concentrations within the above range tends to yield excellent effects in extending healthy lifespan.

[0051] Methods for extending healthy life expectancy do not necessarily have to involve medical procedures.

[0052] [Cell Exposure] The compositions of the present disclosure may be compositions in which cells are exposed to 5 to 100 μM, preferably 10 to 50 μM, of 5-hydroxyequol. Alternatively, the compositions of the present disclosure may be compositions comprising 5-hydroxyequol in a dose that exposes cells to 5 to 100 μM, preferably 10 to 50 μM, of 5-hydroxyequol. These compositions tend to be able to more effectively extend healthy lifespan. The cells may be the cells described above, for example, cells contained in an individual or tissue, or cells isolated from an individual or tissue. If the cells are cells contained in an individual or tissue, 5-hydroxyequol may take the form of a 5 to 100 μM, preferably 10 to 50 μM, 5-hydroxyequol solution within the individual or tissue after administration to the individual or tissue, or after the individual has ingested 5-hydroxyequol. In this case, the 5-hydroxyequol solution may use a body fluid such as blood as the solvent. Therefore, exposure of cells to 5-hydroxyequol in a concentration of 5-100 μM, preferably 10-50 μM, may occur, for example, after an individual has been administered 5-hydroxyequol or a composition containing it, or after an individual has ingested 5-hydroxyequol or a composition containing it, and the cells within the individual's body are exposed to a bodily fluid (e.g., blood) containing 5-hydroxyequol in a concentration of 5-100 μM, preferably 10-50 μM.

[0053] <Other Embodiments> In one embodiment, a method for culturing or preserving cells and / or tissues is disclosed, which includes adding the above-described composition to a culture medium as a cell culture reagent.

[0054] In one embodiment, a culture kit for extending the healthy lifespan of cells and / or tissues is disclosed, comprising the above-described composition.

[0055] In one embodiment, a method for extending healthy life expectancy is disclosed, which includes administering a required amount of 5-hydroxyequol to an individual who requires extension of healthy life expectancy.

[0056] In one embodiment, a method for extending healthy lifespan is disclosed, which includes administering a required amount of 5-hydroxyequol produced from genistein by fermentation to an individual who requires an extension of healthy lifespan.

[0057] In one embodiment, 5-hydroxyequol is disclosed for use in extending healthy life expectancy.

[0058] In one embodiment, 5-hydroxyequol produced from genistein by fermentation is disclosed for use in extending healthy life expectancy.

[0059] In one embodiment, the use of 5-hydroxyequol for the production of the above-described composition is included.

[0060] In one embodiment, the use of 5-hydroxyequol produced from genistein by fermentation for the production of the above-described composition is included.

[0061] In one embodiment, a composition for extending healthy lifespan is disclosed, comprising 5-hydroxyequol produced from genistein by fermentation.

[0062] A non-limiting list of exemplary embodiments and combinations of exemplary embodiments of the present disclosure is disclosed below: [1] A composition comprising 5-hydroxyequol for extending healthy lifespan. [2] The composition according to [1], wherein extending healthy lifespan is to reduce the decline in physical and mental functions of an individual associated with aging. [3] The composition according to [1] or [2], wherein extending healthy lifespan is to reduce the decline in motor functions of an individual associated with aging. [4] The composition according to any one of [1] to [3], wherein extending healthy lifespan is to reduce or support the likelihood of an individual developing frailty, locomotive syndrome, and / or sarcopenia; to improve or support the health of an individual who has developed frailty, locomotive syndrome, and / or sarcopenia; and / or reduce or support the likelihood of an individual who has developed frailty, locomotive syndrome, and / or sarcopenia becoming more unhealthy. [5] The composition according to any one of [1] to [4], wherein extending healthy life expectancy is the prevention of sudden death in young people. [6] The composition according to any one of [1] to [5], administered orally to an individual. [7] The composition according to any one of [1] to [6], which is a food composition. [8] The composition according to any one of [1] to [7], wherein cells are exposed to 5-100 μM of 5-hydroxyequol. [9] A method for producing the composition according to any one of [1] to [8], comprising producing 5-hydroxyequol from genistein by fermentation.

[10] A method for extending healthy life expectancy, comprising contacting a target with 5-hydroxyequol.

[11] The method according to

[10] , wherein cells are exposed to a 5-100 μM solution of 5-hydroxyequol.

[0063] The present disclosure will be further illustrated by the following examples, but these examples will not limit the interpretation of the present disclosure.

[0064] [Example 1] Forced Swimming Behavior Test Wild-type nematode C. elegans strain N2 was cultured in a constant temperature incubator at 20°C. NGM agar plates coated with uracil-requiring Escherichia coli strain OP50, which serves as food for the nematodes, were used as the culture medium. The nematodes hatch from eggs and become reproductively capable adults after 3-4 days. Therefore, subculturing was performed by transferring the nematodes on the above medium to a new medium using a platinum wire every 3-4 days. Subcultured nematodes (L4 stage) were transferred to NGM agar plates coated with uracil-requiring Escherichia coli strain OP50 containing 0 (control group), 10, 30, 50, or 100 μM of 5-hydroxyequol and the oviposition inhibitor 2'-Deoxy-5-fluorouroidine (FUDR), and cultured on the same medium to administer 5-hydroxyequol to the nematodes (Figure 1).

[0065] One week after the start of 5-hydroxyequol administration, nematodes were transferred to M9 medium and allowed to acclimate for one minute. Their movement for one minute was then recorded using a fluorescence microscope, and the number of times the nematodes flapped in the M9 medium (the number of times the nematode's head arced back and forth in 30 seconds) was measured to perform a forced swimming behavior test of the nematodes. The results showed that the groups treated with 10, 30, or 50 μM 5-hydroxyequol did not show a significant change in swimming behavior compared to the control group of nematodes that did not receive 5-hydroxyequol. Nematodes treated with 100 μM 5-hydroxyequol were not analyzed due to the birth of offspring. This result indicates that 10, 30, or 50 μM 5-hydroxyequol does not reduce the individual's motility. Based on these results, it was determined that the evaluation of the ability to extend healthy lifespan using C-HAS in Example 2 should be conducted within the 5-hydroxyequol concentration range of 10–50 μM.

[0066] [Example 2] Evaluation of the ability to extend healthy lifespan using C-HAS The ability of 5-hydroxyequol to extend healthy lifespan was evaluated using C-HAS (C-HAS+ Inc.), an automated technology for measuring healthy lifespan using nematodes. Specifically, 10, 30, or 50 units of 5-hydroxyequol were administered to nematodes (young adult stage, n=30) for one week using the same procedure as described above. After transferring the nematodes to M9 medium, still images were acquired every 12 hours using the live cell imaging analysis instrument IncuCyte® S3 (Essen bio science Inc.) by phase contrast observation and fluorescence observation (excitation light: 440-480 nm, fluorescence: 504-544 nm, exposure time 200 milliseconds) until all nematodes died. The nematodes were reared in an IncuCyte® S3 incubator at 23°C from the administration of 5-hydroxyequol until death. By superimposing the acquired sequential images and observing the degree of overlap of the nematodes (phase difference), active nematodes (active and healthy), immobile nematodes (unhealthy nematodes that remain in place but only their heads and tails move), and dead nematodes (nematodes that are not moving at all (immobile)) were distinguished. Using this distinction as an indicator, in this example, the period from the beginning of the adult stage to the immobile stage was defined as the healthy lifespan, and the period from the beginning of the adult stage to death was defined as the lifespan.

[0067] By defining individuals with a longer healthy lifespan than the average healthy lifespan of the control group as healthy and individuals with a shorter healthy lifespan as unhealthy, and by defining individuals with a longer lifespan than the average lifespan of the control group as long-lived and individuals with a shorter lifespan as short-lived, the nematodes were classified into the following four groups: 1. Healthy, short-lived group ("Short-lived" group) 2. Unhealthy, long-lived group ("Standard" group) 3. Healthy, long-lived group ("Healthy Long-lived" group) 4. Unhealthy, short-lived group ("Unhealthy" group) Based on the above classification, the percentage of each group was calculated as the percentage of the number of individuals in each group relative to the total number of individuals in groups 1 to 4. From the calculated percentages of each group, the quadrant Healthable score (qHaS) was calculated based on the following formula. qHaS = (3. Percentage of healthy, long-lived individuals) - [(4. Percentage of unhealthy, short-lived individuals) + (1. Percentage of healthy, short-lived individuals)]

[0068] As shown in Figure 2, the qHaS of the nematode group treated with 30 μM 5-hydroxyequol was 18.5, while the qHaS of the nematode group treated with 50 μM 5-hydroxyequol was 42.1. This result indicates that 5-hydroxyequol has an effect of effectively extending healthy lifespan.

[0069] Furthermore, as shown in Figure 2, no "1. Healthy, short-lived population (short-lived population)" was observed in the nematode populations to which 30 μM and 50 μM 5-hydroxyequol were added. In other words, these nematode populations tended not to die suddenly at a young age without going through a period of ill health. This result indicates that 5-hydroxyequol effectively prevents sudden death at a young age.

[0070] Furthermore, in analyses using C-HAS, nematodes that are classified as immobile nematodes (unhealthy nematodes that remain in place but only their heads and tails move) are considered to be in a state of decline in physical and mental functions (e.g., motor function) associated with aging, such as frailty, locomotive syndrome, and / or sarcopenia, and the mechanisms involved (e.g., decreased myosin expression, increased mitochondrial fragmentation, decreased motility, etc.) are common to those of mammals. Therefore, the above results indicate that 5-hydroxyequol universally has the effect of mitigating the decline in physical and mental functions (e.g., motor function) of individuals associated with aging in mammals, including humans, and for example, it has the effect of reducing or supporting the likelihood of frailty, locomotive syndrome, and / or sarcopenia occurring in individuals, improving or supporting the health status of individuals who have frailty, locomotive syndrome, and / or sarcopenia, and / or reducing or supporting the likelihood of individuals who have frailty, locomotive syndrome, and / or sarcopenia falling into a more unhealthy state.

[0071] The composition of this embodiment can extend healthy life expectancy and can therefore be suitably used, for example, as a food composition, thus possessing industrial applicability.

Claims

1. A composition containing 5-hydroxyequol for extending healthy lifespan.

2. The composition according to claim 1, wherein extending healthy life expectancy includes mitigating the decline in physical and mental functions of an individual that occurs with aging.

3. The composition according to claim 1, wherein extending healthy life expectancy includes mitigating the decline in an individual's motor function associated with aging.

4. The composition according to claim 1, wherein extending healthy life expectancy reduces or supports the likelihood of an individual developing frailty, locomotive syndrome, and / or sarcopenia; improves or supports the health of an individual who has developed frailty, locomotive syndrome, and / or sarcopenia; and / or reduces or supports the likelihood of an individual who has developed frailty, locomotive syndrome, and / or sarcopenia becoming more unhealthy.

5. The composition according to claim 1, wherein extending healthy life expectancy includes preventing sudden death in young people.

6. The composition according to claim 1, which is administered orally to an individual.

7. The composition according to claim 1, which is a food composition.

8. The composition according to claim 1, wherein cells are exposed to 5-100 μM of 5-hydroxyequol.

9. A method for producing the composition according to any one of claims 1 to 8, comprising producing 5-hydroxyequol from genistein by fermentation.

10. A method for extending healthy life expectancy, comprising contacting 5-hydroxyequol with a target substance.

11. The method according to claim 10, wherein cells are exposed to a 5-100 μM 5-hydroxyequol solution.

Images