JP7633238B2 - Nicotinamide adenine dinucleotide (NAD) concentration increaser - Google Patents

Description

The present invention relates to an agent for increasing nicotinamide adenine dinucleotide (NAD) concentration.

In recent years, in countries with a high elderly population, extending healthy lifespan has become an issue. One of the factors that shorten healthy lifespan is the decline in bodily functions due to the decline in nicotinamide adenine dinucleotide (NAD ⁺ ) production in the body that accompanies aging.

Here, nicotinamide adenine dinucleotide (NAD ⁺ (hereinafter also referred to as NAD)) is a coenzyme that mediates many redox reactions in the body. It is known that NAD levels decrease during the aging process, causing defects in nuclear and mitochondrial function, and causing many age-related pathologies. For example, Non-Patent Document 1 suggests that aging and aging-related diseases are associated with a decrease in intracellular nicotinamide adenine dinucleotide (NAD ⁺ ) levels and a decrease in the enzyme activity of the longevity gene sirtuin, which is an NAD ⁺ -dependent deacetylase. In recent years, it has been reported that increasing NAD ⁺ activates sirtuin and provides an anti-aging effect (Non-Patent Document 2). For this reason, vigorous research is being conducted on the search for components that are effective in preventing or improving the decrease in NAD ⁺ associated with aging and can be applied to medicines, foods for specified health uses, functional foods, etc.

Specifically, Non-Patent Document 2 has shown that the levels of extracellular nicotinamide phosphoribosyltransferase (eNAMPT) decrease with age in mice and humans, and has disclosed that an increase in eNAMPT promotes NAD levels, inhibits aging, and extends lifespan. It has also been suggested that supplementation with extracellular vesicles (EVs) containing eNAMPT improves physical activity and extends lifespan in mice, and thus may be a potential intervention for inhibiting aging.

On the other hand, there has been no suggestion or disclosure regarding the effect of sesamin and/or episesamin in increasing nicotinamide adenine dinucleotide (NAD) in the body.

Imai, S. & Guarente, L. (2014) Trends Cell Biol. , 24, 464-471. “NAD+ and sirtuins in aging and disease” Yoshida M, Satoh A, Lin JB, Mills KF, Sasaki Y, Rensing N, Wong M, Apte RS, Imai SI. , Cell Metab. (2019) 30(2):329-342. e5. “Extracellular Vesicle-Contained eNAMPT Delays Aging and Extends Lifespan in Mice”

The object of the present invention is to provide a safe nicotinamide adenine dinucleotide (NAD) concentration elevating agent derived from food ingredients.

As a result of intensive research conducted by the inventor to solve the above problems, the inventor discovered that sesamin derivatives have the effect of increasing nicotinamide adenine dinucleotide (NAD).

The present invention relates to the following nicotinamide adenine dinucleotide (NAD) concentration elevating agents:
[1] A nicotinamide adenine dinucleotide (NAD) concentration-elevating agent comprising one or more sesamin derivatives as an active ingredient.
[2] The agent according to the above [1], wherein one or more of the sesamin components is sesamin and/or episesamin.
[3] The agent according to [1] or [2] above, which increases, inhibits a decrease in, maintains or improves NAD concentration.
[4] The agent according to any one of [1] to [3] above, which improves, inhibits the decline of, maintains or improves mitochondrial function through an increase in NAD concentration.
[5] The agent according to any one of [1] to [3] above, which enhances, inhibits the decline of, maintains or improves mitochondrial energy production ability through an increase in NAD concentration.
[6] The agent according to any one of [1] to [5] above, which is used for inhibiting and/or delaying aging associated with a decrease in NAD concentration.
[7] The agent according to any one of the above [1] to [6], which is an oral composition.
[8] The agent according to any one of [1] to [7] above, which is a food or drink.
[9] The agent according to any one of [1] to [8] above, which is labeled as "inhibiting and/or delaying aging associated with a decrease in NAD concentration" and/or "inhibiting and/or delaying cellular aging."
[10] A method for increasing nicotinamide adenine dinucleotide (NAD) levels by administering one or more types of sesamin derivatives.
[11] A method for increasing, inhibiting a decrease in, maintaining, or improving nicotinamide adenine dinucleotide (NAD) concentration, which comprises administering one or more types of sesamin derivatives.
[12] Use of one or more sesamin derivatives for increasing nicotinamide adenine dinucleotide (NAD) concentration.
[13] Use of one or more sesamin derivatives for increasing, inhibiting a decrease, maintaining or improving nicotinamide adenine dinucleotide (NAD) concentration.

According to the present invention, a nicotinamide adenine dinucleotide (NAD) concentration increase agent can be provided. Ingestion of one or more types of sesamin analogues can provide the effect of increasing nicotinamide adenine dinucleotide (NAD) concentration. The sesamin analogues used in the present invention are components that have been ingested as foods since ancient times, and are advantageous in that they are highly safe and can be ingested continuously.

1 is a graph showing the effect of a sesamin/episesamin mixture (SE) (sesamin:episesamin (weight ratio = 1:1)) in increasing intracellular NAD levels.

The nicotinamide adenine dinucleotide (NAD) concentration-increasing agent of the present invention contains one or more types of sesamin derivatives.
One or more kinds of sesamin are used to increase nicotinamide adenine dinucleotide (NAD) concentration, and have the effect of increasing, suppressing, maintaining or improving nicotinamide adenine dinucleotide (NAD) concentration, and are used to increase NAD concentration, suppress the decrease in NAD concentration, maintain NAD concentration, or improve NAD concentration.In addition, the agent of the present invention is preferably used to increase, suppress the decrease, maintain or improve NAD concentration that has decreased due to aging.The nicotinamide adenine dinucleotide concentration increasing agent can also be called a nicotinamide adenine dinucleotide (NAD) amount increasing agent.In addition, NAD concentration means the NAD concentration in cells.

Sesamin compounds have the effect of increasing nicotinamide adenine dinucleotide (NAD) concentration, and are expected to contribute to the prevention or improvement of aging associated with a decrease in NAD concentration by increasing, inhibiting, maintaining, or improving NAD concentration. Aging is understood to be a phenomenon in which physical, physiological, and mental functions decline. Physical changes due to aging begin around the age of 40 after reaching maturity, and include wrinkles on the skin, hair and teeth loss, graying of hair, accumulation of fatigue and decreased recovery, decreased eyesight and hearing, decreased motor function, decreased muscle strength and activity, decreased quality of sleep, and decreased bone mass. Although aging itself cannot be considered a disease, the decline in physical and physiological functions increases the risk of so-called geriatric diseases such as arteriosclerosis, glucose and lipid metabolism disorders, neurodegeneration, osteoporosis, and cataracts, and the decline in physical functions also leads to aging of mental functions such as memory and learning. Increasing the concentration of nicotinamide adenine dinucleotide (NAD) is effective in preventing or improving aging associated with a decrease in NAD concentration.

In one aspect, the nicotinamide adenine dinucleotide (NAD) concentration elevating agent of the present invention enhances, inhibits, maintains or improves NAD concentration, and is preferably used for enhancing, inhibiting, maintaining or improving NAD concentration that has decreased due to aging. In one aspect, the agent is preferably used for enhancing, inhibiting, maintaining or improving NAD concentration that has decreased due to aging in middle-aged and elderly people. Middle-aged and elderly people include elderly people. Middle-aged and elderly people may be, for example, people aged 40 years or older, and elderly people may be, for example, people aged 60 years or older or 65 years or older.
The NAD concentration of animals such as humans can be measured by a conventional method in the technical field to which the present invention belongs, for example, by measuring the intracellular NAD concentration using the NAD ⁺ /NADH Quantification Colorimetric Kit (Biovision) or the Amplite (registered trademark) Colorimetric NAD/NADH Ratio Assay Kit (AAT Bioquest).

The nicotinamide adenine dinucleotide (NAD) concentration elevating agent of the present invention can be used for the treatment of conditions or diseases that can be expected to be prevented or improved by increasing, suppressing, maintaining or improving NAD concentration. Examples of such conditions or diseases include the decline of physical function, physiological function, and mental function associated with aging, specifically, for example, the occurrence of skin aging symptoms (wrinkles, sagging, loss of skin elasticity, etc.), dry skin due to aging (decrease in skin moisture retention), skin spots, freckles, and rough skin, the decrease or increase in hormone (growth hormone, thyroid hormone, adrenal cortical hormone, sex hormone, prolactin, antidiuretic hormone, parathyroid hormone, melatonin, etc.) secretion, damage to cells (brain cells, cardiac muscle cells, etc.) by active oxygen, loss of hair and teeth, decreased eyesight and hearing, decreased motor function, decreased bone mass, decreased physical strength, decreased memory, decreased learning ability, decreased immune function, and the occurrence of geriatric diseases.
As used herein, prevention includes preventing onset, delaying onset, reducing the incidence, reducing the risk of onset, etc. Amelioration of a condition or disease includes recovering a subject from a condition or disease, alleviating the symptoms of a condition or disease, ameliorating the symptoms of a condition or disease, delaying or preventing the progression of a condition or disease, etc.

In the present invention, sesamin analogs are a general term for compounds including sesamin and its analogs. Sesamin is one of the main lignan compounds contained in sesame. In addition to episesamin, examples of sesamin analogs include dioxabicyclo[3.3.0]octane derivatives described in JP-A-4-9331. As one or more types of sesamin, one of these compounds may be used alone, or two or more types may be used. Specific examples of sesamin analogs include sesamin, episesamin, sesaminol, episesaminol, sesamol, sesamolin, etc., and these stereoisomers or racemates may be used alone or in mixture. In addition, metabolites of sesamin (for example, as described in JP-A-2001-139579) are also sesamin analogs contained in the sesamins of the present invention as long as they exhibit the effects of the present invention, and can be used in the present invention. In the present invention, sesamin and/or episesamin can be preferably used as one or more of the sesamins, and sesamin and episesamin can be more preferably used. When sesamin and episesamin are used, the ratio thereof is not particularly limited, but for example, the sesamin:episesamin (weight ratio) is preferably 1:0.1 to 1:9, more preferably 1:0.3 to 1:3, and even more preferably 1:0.5 to 1:2.

The sesamin compounds used in the present invention are not limited in any way by their form or manufacturing method. For example, in the case of sesamin, sesamin (called sesamin extract or purified product) extracted from sesame oil by a known method (for example, the method described in JP-A-4-9331) can be used. Commercially available sesame oil (liquid) can also be used as is. However, when sesame oil is used, the flavor peculiar to sesame oil may be evaluated as being undesirable from an organoleptic perspective, so it is preferable to use a tasteless and odorless sesamin extract (or sesamin purified product) extracted from sesame oil. In addition, when sesame oil is used, the sesamin content is low, so if a desired amount of sesamin is to be incorporated, the volume per unit dose of the formulated composition becomes too large, which may cause inconvenience in ingestion. In particular, when formulated for oral administration, the formulation (tablet, capsule, etc.) becomes too large, which causes difficulties in ingestion. Therefore, it is preferable to use a sesamin extract (or sesamin purified product) from sesame oil from the viewpoint of small intake amounts. Sesamin compounds can also be obtained by synthesis. For example, sesamin and episesamin can be synthesized by the method of Beroza et al. (J. Am. Chem. Soc., 78, 1242 (1956)). Metabolites of sesamin and episesamin can be synthesized by the method of Urata et al. (Chem. Pharm. Bull. (Tokyo), 56 (11) 1611-2 (2008)).

Sesamin compounds are found in natural products and foods and beverages, have a wide dietary history, and are recognized for their high safety. Sesamin compounds, with their guaranteed safety, are ideal for continuous and long-term ingestion. According to the present invention, it is possible to provide a nicotinamide adenine dinucleotide (NAD) concentration elevating agent that contains a highly safe substance as an active ingredient.

NAD in the body plays an important role in promoting the expression and synthesis of molecules involved in mitochondrial function and mitochondrial energy production by activating sirtuins and deacetylating downstream enzymes or transcription factors. Therefore, an increase in NAD concentration can improve, inhibit the decline, maintain or improve mitochondrial function. The agent of the present invention can improve, inhibit the decline, maintain or improve mitochondrial function through an increase in NAD concentration. Furthermore, the agent of the present invention can improve, inhibit the decline, maintain or improve mitochondrial energy production ability through an increase in NAD concentration. This also allows the agent of the present invention to have an anti-cellular aging effect.

In this specification, mitochondrial function and mitochondrial energy production capacity may be evaluated based on common knowledge in the technical field to which the present invention belongs, and the method is not particularly limited. For example, mitochondrial function can be evaluated by continuously measuring the oxygen consumption rate (hereinafter also referred to as "OCR") used by mitochondria during ATP synthesis in the cells to be measured using an extracellular flux analyzer while adding an ATP synthase inhibitor (e.g., oligomycin and rotenone) and an uncoupler (e.g., carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP)). Mitochondrial function can be evaluated by analyzing, for example, the basal respiration rate, ATP production capacity, and maximum respiration rate, which are considered to be major indicators in the evaluation of mitochondrial function.

The agents of the present invention can be used to inhibit and/or delay aging associated with a decrease in NAD concentration.

The agent of the present invention can be applied to either therapeutic use (medical use) or non-therapeutic use (non-medical use). Non-therapeutic use is a concept that does not include medical procedures, i.e., surgery, treatment or diagnosis of humans.
The agent of the present invention can be in the form of a food or drink, a medicine, a quasi-drug, a feed, etc. The agent of the present invention may be a food or drink, a medicine, a quasi-drug, a feed, etc. that is used for increasing the concentration of nicotinamide adenine dinucleotide (NAD) or for increasing, suppressing a decrease, maintaining, or improving the NAD concentration, or may be a material or preparation that is mixed with these and used.
Furthermore, the agent of the present invention may be a food or drink, medicine, quasi-drug, feed, etc., which is intended to improve, suppress the decline, maintain or improve mitochondrial function through an increase in NAD concentration, or to improve, suppress the decline, maintain or improve mitochondrial energy production ability through an increase in NAD concentration, or may be a material or preparation, etc., which is mixed with these and used.
The agent of the present invention can be provided in the form of an agent as it is, but is not limited to this form. The agent can also be provided as a composition as it is, or as a composition containing the agent.
The agent of the present invention may be either an oral agent or a parenteral agent, but is preferably an oral agent. The oral agent may be a food or drink, an oral medicine, a quasi-drug, a feed, etc., or may be included in these. The agent of the present invention is preferably a food or drink or an oral medicine, and more preferably a food or drink.

The agent of the present invention may contain any additives and any components in addition to one or more types of sesamin-class compounds, so long as the effects of the present invention are not impaired. These additives and components may be selected depending on the form of the agent or composition, and those that can generally be used in foods and beverages, medicines, quasi-drugs, feed, etc. may be used. When the agent of the present invention is used as a food and beverage, medicine, quasi-drug, feed, etc., there are no particular limitations on the manufacturing method, and it may be manufactured by a general method.

For example, when the agent of the present invention is made into a food or beverage, one or more types of sesamin-class compounds can be mixed with ingredients that can be used in food or beverage (e.g., food ingredients, food additives used as necessary, etc.) to make various foods and beverages. The foods and beverages are not particularly limited, and examples include general foods and beverages, health foods, health drinks, functional foods, foods for specified health uses, and foods and beverages for sick people. The above health foods, functional foods, 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, and liquid diets.

When the agent of the present invention is used as a drug or quasi-drug, for example, one or more types of sesamin-class compounds can be mixed with a pharmacologically acceptable carrier and additives added as necessary to produce a drug or quasi-drug in various dosage forms. Such carriers, additives, etc. may be any pharmacologically acceptable carriers that can be used in drugs or quasi-drugs, and examples of such carriers, additives, etc. include one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, etc. Examples of the administration (ingestion) form of the drug or quasi-drug include oral or parenteral (transdermal, transmucosal, enteral, injection, etc.) administration forms. When the agent of the present invention is used as a drug or quasi-drug, it is preferable to use an oral drug or quasi-drug. Examples of dosage forms for oral administration include liquids, tablets, powders, fine granules, granules, sugar-coated tablets, capsules, suspensions, emulsions, chewables, etc. The drug may be a drug for non-human animals.

When the agent of the present invention is used as feed, one or more types of sesamin compounds may be blended into the feed. Feed also includes feed additives. Examples of feed include livestock feed for cows, pigs, chickens, sheep, horses, etc.; small animal feed for rabbits, rats, mice, etc.; and pet food for dogs, cats, small birds, etc.

The amount of sesamin-class compounds contained in the agent of the present invention is not particularly limited and can be set depending on the form, etc.
The total content of sesamin-class compounds in the agent of the present invention is, for example, preferably 0.001% by weight or more in the agent, more preferably 0.01% by weight or more, even more preferably 0.05% by weight or more, and preferably 10% by weight or less, more preferably 5% by weight or less. In one embodiment, the total content of sesamin-class compounds in the agent 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. When two or more types of sesamin-class compounds are contained, the above total content is the total content of these compounds.

The agent of the present invention is preferably an oral composition (a composition that is taken orally (administered orally)). The dosage (which can also be referred to as the intake amount) of the agent of the present invention is not particularly limited. The dosage of the agent of the present invention may be any amount that can obtain the effect of increasing nicotinamide adenine dinucleotide concentration, or the effect of improving, inhibiting a decrease, maintaining or improving NAD concentration, and may be set appropriately depending on the dosage form, administration method, subject's body weight, etc.

In one aspect, when the agent of the present invention is orally ingested or administered to a human (adult), the total dosage of sesamin-based compounds 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 per 60 kg body weight. In one aspect, the total dosage of sesamin-based compounds is preferably 0.5 to 200 mg, more preferably 1 to 100 mg, and even more preferably 3 to 80 mg per day per 60 kg body weight for a human (adult). It is preferable to ingest or administer the above amount once or more times a day, for example, once a day or in divided doses several times a day (for example, 2 to 3 times). In one aspect, it is preferable to orally ingest or administer the above amount of sesamin-based compounds to a human. In one aspect, the agent of the present invention can be used to ingest or administer the above amount of sesamin-based compounds per day per 60 kg body weight to a human. When two or more sesamin-class compounds are used, the total dosage is the combined amount. In one embodiment, sesamin and/or episesamin is orally ingested or administered to a human (adult) at a total dosage 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 per 60 kg body weight.

It is preferable that the agent of the present invention is taken or administered continuously. It is expected that the above-mentioned effects will be enhanced by the continuous intake or administration of sesamin-related compounds. In one aspect, it is preferable that the agent of the present invention is taken or administered continuously, preferably for one week or more, more preferably for four weeks or more, even more preferably for eight weeks or more, and particularly preferably for twelve weeks or more.

The nicotinamide adenine dinucleotide (NAD) concentration increasing agent of the present invention may be labeled with a function exhibited by increasing the nicotinamide adenine dinucleotide (NAD) concentration, or a function exhibited by increasing, suppressing a decrease, maintaining, or improving the NAD concentration. For example, one or more functions selected from the group consisting of "improving, suppressing a decrease, maintaining, or improving mitochondrial function via increasing the NAD concentration", "improving, suppressing a decrease, maintaining, or improving the energy production ability of mitochondria via increasing the NAD concentration", "suppressing and/or delaying aging due to decreasing the NAD concentration", "suppressing aging via increasing the NAD concentration", and "suppressing and/or delaying cellular aging".
In one aspect of the present invention, the agent of the present invention is preferably a food or drink labeled with the above-mentioned label. The above-mentioned label may be a label indicating that the agent is used to obtain the above-mentioned function.

The subject to which the agent of the present invention is ingested or administered (which can also be called the administration subject) is not particularly limited, and the agent can be ingested by humans and non-human animals. Examples of non-human animals include industrial animals, pets, and laboratory animals. Specifically, industrial animals refer to animals that need to be raised industrially, such as livestock such as cows, horses, pigs, goats, and sheep, poultry such as chickens, ducks, quails, turkeys, and ostriches, and fish such as yellowtail, yellowtail, red sea bream, horse mackerel, carp, rainbow trout, and eels. Pets refer to so-called pets and companion animals such as dogs, cats, marmosets, small birds, and hamsters, and laboratory animals refer to animals used in research in the fields of medicine, biology, agriculture, and pharmacy, such as mice, rats, guinea pigs, beagles, miniature pigs, rhesus monkeys, and cynomolgus monkeys.

The subject to which the agent of the present invention is ingested or administered (which may also be referred to as an administration subject) is preferably a human or a non-human mammal, more preferably a human.
In one embodiment, the subjects to be administered include subjects who need or want to increase the nicotine adenine dinucleotide (NAD) concentration; subjects who need or want to improve, suppress, maintain or improve the NAD concentration. Examples of such subjects include middle-aged and elderly people, subjects who need or want to improve, suppress, maintain or improve mitochondrial function through an increase in NAD concentration, subjects who need or want to improve, suppress, maintain or improve the energy production ability of mitochondria through an increase in NAD concentration, and subjects who need or want to suppress and/or delay cellular aging through an increase in NAD concentration. Middle-aged and elderly people may be, for example, humans aged 40 years or older. In one embodiment, among middle-aged and elderly people, elderly people are preferred as subjects. Elderly people may be, for example, humans aged 60 years or older or 65 years or older. The agent of the present invention can also be used for healthy people for the purpose of preventing symptoms or diseases that can be expected to be prevented or improved by increasing the nicotinamide adenine dinucleotide (NAD) concentration, or by improving, suppressing, maintaining or improving the NAD concentration.

The present invention also encompasses the following methods.
A method for increasing nicotinamide adenine dinucleotide (NAD) levels by administering one or more sesamin derivatives.
A method for increasing, inhibiting a decrease, maintaining or improving NAD concentration by administering one or more types of sesamin derivatives.
A method for increasing, inhibiting a decrease, maintaining or ameliorating NAD concentration that has decreased with age, which comprises administering one or more types of sesamin derivatives.

The present invention also encompasses the following uses:
Use of one or more sesamin derivatives for increasing nicotinamide adenine dinucleotide (NAD) concentration.
Use of one or more sesamin derivatives for increasing, inhibiting a decrease, maintaining or improving NAD concentration.
Use of one or more sesamin-class compounds for increasing, inhibiting, maintaining or improving NAD concentration that has decreased with age.
The above methods and uses may be therapeutic or non-therapeutic methods or uses.
By administering one or more kinds of sesamin, it is possible to increase the concentration of nicotinamide adenine dinucleotide (NAD). That is, it is possible to improve, suppress, maintain or improve the NAD concentration. It is also possible to improve, suppress, maintain or improve the NAD concentration that has decreased due to aging. This makes it possible to improve, suppress, maintain or improve mitochondrial function through the increase in NAD concentration, improve, suppress, maintain or improve the energy production ability of mitochondria through the increase in NAD concentration, and suppress and/or delay cellular aging through the increase in NAD concentration.

In the above method and use, the sesamin compounds and their preferred aspects are the same as those of the agent of the present invention described above. As the one or more types of sesamin compounds, one type of sesamin compound may be used, or two or more types may be used. In the above method and use, it is preferable to administer (have the subject ingest) one or more types of sesamin compounds to the subject at least once a day, for example, once to several times (for example, two to three times) a day. In the above method and use, it is preferable to administer (have the subject ingest) the sesamin compounds orally. The above use is preferably in humans or non-human mammals, more preferably in humans.

In the above-mentioned methods and uses, one or more types of sesamin compounds may be used in an amount (which may also be called an effective amount) that provides the desired effect (improvement, inhibition of decline, maintenance or improvement of mitochondrial function, improvement of mitochondrial energy production ability, inhibition of decline or maintenance, and/or inhibition and/or delay of cellular aging due to an increase in nicotinamide adenine dinucleotide (NAD) concentration). The preferred dosage and administration subjects of the sesamin compounds are the same as those of the agent or composition of the present invention described above. The sesamin compounds may be administered as they are, or as an agent or composition containing them. For example, the agent or composition of the present invention described above may be used.

The present invention also includes the use of one or more sesamin derivatives for producing the agent or composition of the present invention.

The present invention will be described in more detail below with reference to examples. Note that the present invention is not limited to these examples.

<Examples 1 and 2, Comparative Example 1: Evaluation test of intracellular NAD ⁺ concentration (NAD concentration) using sesamin/episesamin mixture (SE) (sesamin:episesamin (weight ratio = 1:1))>
To examine the effect on intracellular NAD ⁺ concentration, 1.0 x ¹⁰⁵ cells/well of HepG2 cells were seeded in a 12-well plate and cultured in DMEM medium (Nacalai Tesque, Inc., containing 10% FBS) for 24 hours at 37°C and _CO2 (5%). After 24 hours of culture, the medium in each well was replaced with DMEM medium without sesamin-episesamin mixture (containing 1% albumin, no FBS (Comparative Example 1: control)), DMEM medium containing 1 μM SE (containing 1% albumin, no FBS (Example 1)), or DMEM medium containing 10 μM SE (containing 1% albumin, no FBS (Example 2)), and cultured for another 24 hours. Thereafter, the supplemented medium was removed, the cells in the wells were washed with D-PBS (Nacalai Tesque, Inc.), and then NADH/NAD Extraction Buffer (NAD ⁺ /NADH Quantification Colorimetric Kit, Biovision) was added to recover the cell suspension. Then, a cell extract was obtained according to the Kit manual, and the intracellular NAD ⁺ concentration was analyzed. The intracellular NAD ⁺ concentration was calculated as a value per protein concentration by measuring the protein concentration of the cell extract (Pierce (registered trademark) BCA Protein Assay Kit, Thermo Scientific). The results obtained (average of N=2) are shown in FIG. 1.

1, the intracellular NAD ⁺ concentration was increased by 20% in the SE 1 μM group in Example 1 and 37% in the SE 10 μM group in Example 2 compared to the control group (medium not containing SE) in Comparative Example 1. From the above, the dose-dependent increase in intracellular NAD ⁺ concentration by SE treatment was confirmed.

Claims (9)

Contains sesamin and/or episesamin as active ingredients.
Nicotinamide adenine dinucleotide (NAD) concentration increasing agent. The agent according to claim 1 , which increases, inhibits a decrease in, maintains or improves NAD concentration. The agent according to claim 1 or 2, for preventing or improving skin blemishes, freckles, hair loss, tooth loss, decreased eyesight, decreased hearing or decreased immune function mediated by an increase in NAD concentration. The agent according to any one of claims 1 to 3 , which is an oral composition. The agent according to any one of claims 1 to 4 , which is a food or drink. A method for increasing nicotinamide adenine dinucleotide (NAD) levels by administering sesamin and/or episesamin (excluding medical treatment on humans) . A method for increasing, inhibiting a decrease, maintaining or improving nicotinamide adenine dinucleotide (NAD) concentration by administering sesamin and/or episesamin (excluding medical procedures on humans) . Use of sesamin and/or episesamin for increasing nicotinamide adenine dinucleotide (NAD) levels (excluding medical treatment for humans) . Use of sesamin and/or episesamin for increasing, inhibiting a decrease, maintaining or improving nicotinamide adenine dinucleotide (NAD) concentration (excluding medical treatment for humans) .

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