Composition for increasing amounts of intracellular ATP and intracellular nad

Abstract

The present invention addresses the problem of providing: a composition that increases the amount of intracellular ATP and the amount of intracellular NAD; and a composition that improves motor performance. More specifically, the present invention provides a composition for increasing the amounts of intracellular adenosine triphosphate (ATP) and intracellular nicotinamide adenine dinucleotide (NAD), the composition being obtained by combining (a) and (b): (a) an NAD precursor or a pharmaceutically acceptable salt thereof; and (b) one or more substances selected from the group consisting of uridine and uridine precursors, or one or more pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or one or more pharmaceutically acceptable salts thereof.　Furthermore, the present invention provides a composition for enhancing muscle strength, the composition being characterized by inclluding a combination of (c) and (d): (c) one or more substances selected from the group consisting of β-alanine and β-alanine precursors, or one or more pharmaceutically acceptable salts thereof; and (d) one or more substances selected from the group consisting of uridine and uridine precursors, or one or more pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or one or more pharmaceutically acceptable salts thereof.

Description

Composition for increasing the amount of intracellular ATP and intracellular NAD

[0001] The present invention relates to a composition for increasing the amount of intracellular ATP and intracellular NAD.

[0002] It is well known that adenosine triphosphate (ATP) functions as an energy source in the body and is used in important life activities. Nicotinamide adenine dinucleotide (NAD) + / NADH) is a substance that functions as a coenzyme for dehydrogenase in the body, and its oxidized form (NAD + NADH exists in two forms: oxidized and reduced (NADH) (hereinafter, when there is no distinction between the oxidized and reduced forms, it will be referred to as "NAD"). NADH is converted to NAD in the electron transport complex in mitochondria, which then synthesizes ATP. Therefore, both ATP and NAD play very important roles as energy sources in living organisms or as substances for generating energy. It is known that a decrease in the amount of ATP and NAD in living organisms can cause various diseases. For example, it has been reported that a decrease in ATP and NAD in brain or muscle tissue due to aging increases the risk of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and spinocerebellar degeneration (Non-Patent Documents 1 and 2), as well as sarcopenia (Non-Patent Documents 3 and 4).

[0003] Based on the above, enhancing ATP and NAD is thought to be effective in preventing or ameliorating pathologies thought to be caused by a decrease in the levels of ATP and NAD in the body. To date, a drug combining a xanthine oxidoreductase (XOR) inhibitor with inosine, inosinic acid, or hypoxanthine has been reported as a drug for increasing intracellular ATP levels (Patent Document 1). Furthermore, a composition for increasing NAD levels has been reported that contains two or more selected from an NAD precursor, an upregulator of NAMPT (nicotinamide phosphoribosyl transferase), an upregulator of NQO1 (NAD(P)H quinone dehydrogenase 1), and a downregulator of NNMT (nicotinamide N-methyltransferase) (Patent Document 2). Furthermore, it has been reported that NMN (nicotinamide mononucleotide) and NR (nicotinamide riboside) increase intracellular ATP and NAD levels (Non-Patent Document 6).

[0004] Since ATP and NAD are energy sources in living tissues, it was expected that increasing the amount of these substances in cells would improve athletic performance. For example, administration of inosine, which has been reported to increase ATP levels, was expected to improve athletic performance, but it has been reported that administration of inosine alone does not have the effect of enhancing muscle movement (Non-Patent Document 5). Therefore, it is thought that the mechanism by which the athletic performance of a living organism improves is not as simple as simply increasing the amount of ATP as an energy source.

[0005] Many compositions have been reported to increase the amount of ATP and NAD in the body, but the actual extent of their effect is unclear for many of them, and their effect on enhancing biological functions (e.g., muscle performance) is also unclear for many of them.

[0006] WO2017 / 033963WO2019 / 175587

[0007] Blaszczyk, Metabolites, 10:450 2020, doi:10.3390 / metabo10110450 Li et al., Neurosci. Bull. 40:218-240 2024, https: / / doi.org / 10.1007 / s12264-023-01072-3 Goody and Henry, Skeletal Muscle, 8:9 2018, https: / / doi.org / 10.1186 / s13395-018-0154-1 Abboudi, The Science Journal of the Lander College of Arts and Sciences 12:2 2019, https: / / touroscholar.touro.edu / sjlcas / vol12 / iss2 / 10 McNaughton et al., Int J Sport Nutr. 9:333-344 1999 Sato et al., 25th Annual Meeting of the Japan Society for Food Engineering (2024), August 19, 2024 - August 20, 2024, Presentation Number: p13 (Poster Presentation) and Abstract

[0008] In view of the above circumstances, an objective of the present invention is to provide a composition that increases the amount of intracellular ATP and the amount of intracellular NAD.Furthermore, an objective of the present invention is to provide a composition that increases the amount of intracellular ATP and further improves the athletic performance of muscles.

[0009] As a result of extensive research aimed at solving the above-mentioned problems, the present inventors have found that the administration of a combination of an NAD precursor (e.g., nicotinamide riboside) and uridine and / or inosine to cells significantly increases intracellular ATP and NAD concentrations. To increase intracellular ATP and NAD levels, the present inventors considered supplementing cells with phosphoribosyl pyrophosphate (PRPP), the rate-limiting substance in the intracellular salvage pathway, to activate the pathway. However, because PRPP contains a phosphate group, it is difficult for extracellular PRPP supplementation to activate the salvage pathway. Therefore, the present inventors investigated methods other than supplementing PRPP itself and found for the first time that administration of uridine, a non-purine base derived from pyrimidine nucleosides, activates the salvage pathway. This result is thought to be due to the supply of PRPP by uridine once it enters the cell. Furthermore, in order to identify factors necessary for improving muscle function through increased ATP concentration in vivo, the present inventors found that administering β-alanine to mice for several days followed by administration of inosine improved muscle endurance. The present invention was completed based on these findings.

[0010] That is, the present invention relates to the following (1) to (13): (1) A composition for increasing the amount of intracellular adenosine triphosphate (ATP) and intracellular nicotinamide adenine dinucleotide (NAD), comprising a combination of the following (a) and (b): (a) an NAD precursor or a pharmaceutically acceptable salt thereof, (b) one or more substances selected from the group consisting of uridine and uridine precursors or pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors or pharmaceutically acceptable salts thereof. (2) The composition according to (1) above, wherein the NAD precursor is nicotinamide mononucleotide (NMN) and / or nicotinamide riboside (NR). (3) The composition according to (1) or (2) above, wherein (b) is one or more substances selected from the group consisting of uridine and uridine precursors or pharmaceutically acceptable salts thereof. (4) The composition according to (1) or (2) above, wherein (b) is one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof. (5) The composition according to (1) or (2) above, further comprising a SIRT1 activator. (6) The composition according to (5) above, wherein the SIRT1 activator is a citrus peel extract. (7) The composition according to (6) above, wherein the citrus fruit is calamandarin, shikuwasa, and / or shiranui. (8) The composition according to (5) above, wherein the SIRT1 activator is 3',4',3,5,6,7,8-heptamethoxyflavone, 5-demethylnobiletin, and / or tangeretin. (9) The composition according to (1) or (2) above, characterized in that it is a pharmaceutical composition, a food or beverage composition, or a cosmetic composition. (10) The composition according to (5) above, which is a pharmaceutical composition, a food or drink composition, or a cosmetic composition. (11) A composition for enhancing muscle strength, which is a combination of the following (c) and (d):(c) one or more substances selected from the group consisting of β-alanine and β-alanine precursors, or pharmaceutically acceptable salts thereof; (d) one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof. (12) The composition according to (11) above, wherein the β-alanine precursor is one or more selected from the group consisting of carnosine, anserine, and balenine. (13) The composition according to (11) or (12) above, characterized in that it is a pharmaceutical composition, a food or beverage composition, or a cosmetic composition. In this specification, the symbol "to" indicates a numerical range including the values ​​on either side of it.

[0011] The compositions according to embodiments of the present invention have the effect of increasing the amounts of ATP and NAD in the body, thereby improving blood flow and organ and muscle function, and are effective in maintaining health as well as preventing or treating various diseases in which a decrease in ATP or NAD forms part of the pathology. Furthermore, muscle-strengthening agents and compositions containing such agents according to other embodiments of the present invention are effective in improving muscle strength loss associated with aging, etc., as well as preventing or treating diseases such as sarcopenia and frailty.

[0012] Figure 1 shows the effects of various nucleosides on intracellular ATP and NAD levels. + The results show the effect of the combined use of inosine, NR, and / or uridine on the intracellular ATP and NAD levels. The figures show the levels of each nucleotide in HEK293 cells cultured for 6 hours in a test medium containing glucose-free medium to which each compound was added at a final concentration of 100 μM. Data are expressed as mean ± SD. n=4. The horizontal line in each graph indicates the value for the control group. NR is an abbreviation for nicotinamide riboside (the same applies to other figures). Figure 2 shows the effect of the combined use of inosine, NR, and / or uridine on the intracellular ATP and NAD levels. +These figures show the effects of inosine and hypoxanthine on intracellular ATP levels in HEK293 cells cultured for 6 hours in glucose-free medium containing each compound at a final concentration of 100 μM. Data are presented as mean ± SD. n = 4. p < 0.05 was considered significant and is indicated by an alphabet. The horizontal line in each graph indicates the value for the control group. Figure 3 compares the effects of inosine and hypoxanthine on intracellular ATP levels in HEK293 cells cultured for 1 or 6 hours in glucose-free medium containing each compound at a final concentration of 100 μM. Data are presented as mean ± SD. n = 3, except for the 6-hour control and Hx groups, where n = 2. ** p < 0.01, * p < 0.05. The horizontal line in each graph indicates the value for the control group. Hx is an abbreviation for hypoxanthine. Figure 4. Intracellular NAD + Regarding the effect of enhancing the amount of NAD + The results show the comparison of precursors. NAD in HEK293 cells cultured for 3 hours in a test medium containing 0.1% glucose and each compound added to a final concentration of 100 μM. + The data are expressed as mean ± SD. n=3. p<0.05 was considered a significant difference and is indicated by alphabetical characters. The horizontal line on the graph indicates the value of the control group. NMN is an abbreviation for nicotinamide mononucleotide, NA is an abbreviation for nicotinic acid, and NAM is an abbreviation for nicotinamide. Figure 5 shows the intracellular ATP and NAD + The graph shows the results of MS measurements of each nucleotide in HEK293 cells cultured for a certain period of time in a test medium containing glucose-free medium to which a stable isotope had been added (final concentrations are shown in the graph). A shows cells at 1 mM 13 C6: Graph showing the ratio of m / Z peak intensity over time after incubation in a medium containing glucose. B and C show the ratio of m / Z peak intensity over time for ATP and NAD, respectively. + This graph shows the ratio of the intensity of the peak with a m / z shift of +5 over time. n=1. Figure 6 shows the intracellular ATP and NAD +The results show that ATP and NAD in HEK293 cells cultured for a certain period of time in a glucose-free medium or a high glucose (0.45%) medium to which a stable isotope was added at a final concentration of 100 μM were measured. + The MS measurement results are shown below. A is the 13 The structure of C5 inosine is shown. B and C are ATP and NAD, respectively. + This graph shows the percentage of the intensity of the peak with a m / z shift of +5 over time. Data are shown as mean ± SD. n=3. **p<0.01, *p<0.05. Figure 7 shows the intracellular ATP and NAD + The results show that ATP and NAD in HEK293 cells cultured for a certain period of time in a glucose-free medium or a high glucose (0.45%) medium to which a stable isotope was added at a final concentration of 100 μM were measured. + The MS measurement results are shown below. A is the 13 The structure of C5 uridine is shown. B and C are ATP and NAD, respectively. +This graph shows the percentage of the m / z +5 shift in peak intensity over time. Data are shown as mean ± SD. n = 3, except for 6-hour glucose-free and 3-hour high-glucose conditions (n ​​= 2). **p<0.01, *p<0.05. Figure 8 shows the effect of a uridine phosphorylase inhibitor on the incorporation of uridine-derived ribose into intracellular ATP. The results of MS analysis of ATP in HEK293 cells cultured for 6 hours in glucose-free medium supplemented with a stable isotope at a final concentration of 100 μM are shown. The percentage of the m / z +5 shift in peak intensity is shown as a graph. Data are shown as mean ± SD. n = 3. **p<0.01, *p<0.05. Figure 9 shows the effects of 9 consecutive days of β-alanine administration and a single inosine administration on muscle endurance using a treadmill. The horizontal axis represents running time, and the vertical axis represents the number of electrical stimulations. Figure 10 shows the results of a treadmill study to examine the effects of two consecutive weeks of β-alanine administration and a single dose of inosine on muscle endurance in mice. The horizontal axis represents running time, and the vertical axis represents the number of electrical stimulations.

[0013] Hereinafter, embodiments of the present invention will be described. The term "present embodiment" refers to all embodiments described herein unless otherwise specified. A first embodiment is a composition for increasing the amount of intracellular adenosine triphosphate (ATP) and intracellular nicotinamide adenine dinucleotide (NAD), comprising a combination of the following: (a) an NAD precursor or a pharmaceutically acceptable salt thereof; (b) one or more substances selected from the group consisting of uridine and uridine precursors or a pharmaceutically acceptable salt thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors or a pharmaceutically acceptable salt thereof. In the first embodiment, "combining (a) and (b)" refers to all aspects of administering component (a) and component (b) so as to increase the amount of ATP and NAD in the body (or cells) of a subject to which the composition is administered. That is, the composition according to the first embodiment is not particularly limited, but may be, for example, a combination drug in which component (a) and component (b) are mixed to form a composition, or may be a drug in which component (a) and component (b) are prepared separately but are provided to be administered at the same time. In this specification, "administered at the same time" includes not only simultaneous administration (for example, administration of a combination drug), but also administration of each component at an interval that allows the components to exert the effect of increasing ATP and NAD levels.

[0014] In this embodiment, component (a) is an NAD precursor or a pharmaceutically acceptable salt thereof. Here, the term "NAD precursor" refers to a compound that converts NAD (NAD +The term "NAD precursor" refers to a substance that is converted into NAD precursor (NADH), and includes, but is not limited to, NAD, nicotinamide adenine dinucleotide phosphate (NADP), nicotinamide mononucleotide (NMN), nicotinamide riboside (NR), nicotinamide (NAM), nicotinic acid (NA), 1-methylnicotinamide (MNA), nicotinic acid adenine dinucleotide (NaAD), nicotinic acid mononucleotide (NaMN), etc. Component (a) may contain multiple NAD precursors (e.g., NMN and NR).

[0015] In the first embodiment, component (b) is one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof. Component (b) may contain one or more substances. That is, for example, component (b) may contain only uridine, only inosine, or uridine and inosine. For further example, component (b) may contain uridine and / or one or more uridine precursors, and / or inosine and / or one or more inosine precursors. Here, the term "uridine precursor" refers to a substance that is converted into uridine through one or more steps within a cell, and includes, but is not limited to, orotidine monophosphate, uridine monophosphate, uridine diphosphate, uridine triphosphate, uridine diphosphate glucose, uridine diphosphate galactose, uridine diphosphate glucosamine, orotic acid, dihydroorotate, triacetyluridine, N-carbamoyl aspartate, cytidine, cytidine monophosphate, cytidine diphosphate, cytidine triphosphate, and the like.

[0016] Furthermore, the term "inosine precursor" refers to a substance that is converted to inosine through one or more steps within a cell, and includes, but is not limited to, hypoxanthine, inosine monophosphate, inosine diphosphate, inosine triphosphate, adenosine monophosphate, adenosine diphosphate, adenosine triphosphate, guanosine monophosphate, guanosine diphosphate, guanosine triphosphate, and xanthosine monophosphate. The use of uridine, a uridine precursor, or a pharmaceutically acceptable salt thereof as component (b) can reduce the amount of inosine, a purine, used.

[0017] The composition according to the first embodiment may further contain a SIRT1 activator in addition to the component (a) and the component (b). +SIRT1 is a protein-dependent deacetylase encoded by Sirt1, a member of the Sirtuin family known as longevity genes. SIRT1 was initially discovered as a deacetylase that removes acetyl groups from lysine residues on histones, but it is also known to have many substrates other than histones, including p53, peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), and FoxO. In this embodiment, the SIRT1 activator is not particularly limited, but examples thereof include extracts from citrus peels (e.g., calamandarin peel, shikuwasa peel, and shiranui (Dekopon (registered trademark) peel)). More specifically, examples thereof include 3',4',3,5,6,7,8-heptamethoxyflavone (CAS number: 1178-24-1), 5-demethylnobiletin (CAS number: 2174-59-6), and tangeretin (CAS number: 481-53-8) contained in these citrus peels. Further examples of the SIRT1 activator include resveratrol, quercetin derived from black turmeric, and the like. 3,5,7,3',4'-pentamethyl ether (CAS number: 1247-97-8) may also be used.

[0018] A second embodiment is a composition for enhancing muscle strength, characterized by combining the following (c) and (d): (c) one or more substances selected from the group consisting of β-alanine and β-alanine precursors, or pharmaceutically acceptable salts thereof; (d) one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof. In the second embodiment, "combining (c) and (d)" means that components (c) and (d) are prepared separately and provided so that they are administered at different times. The order of administration of components (c) and (d) and the number of administrations of each component are not particularly limited. For example, either component (c) or component (d) may be administered once or multiple times, or either component (c) or component (d) may be administered first. Preferably, component (c) may be administered multiple times followed by a single administration of component (d). More specifically, for example, component (c) may be administered daily for 7 to 10 days, followed by a single administration of component (d). In this embodiment, "muscle strength" refers to the force exerted by a muscle in a single contraction and the muscle's ability to continue repeated contractions, and is a concept that encompasses both maximum muscle strength (e.g., assessed by the maximum weight that can be lifted in one go) and muscle endurance (e.g., assessed by the number of times that a load can be sustained). Furthermore, "β-alanine precursor" refers to a substance that is converted into β-alanine via one or more steps within cells. Examples include, but are not limited to, carnosine, anserine, and balenine. Component (c) may contain one or more substances. That is, for example, component (c) may contain only β-alanine, one or more types of β-alanine precursors, or β-alanine and one or more types of β-alanine precursors.For example, component (c) may contain only β-alanine, or may contain one or more substances selected from the group consisting of carnosine, anserine, and balenine. Alternatively, component (c) may contain, in addition to β-alanine, one or more substances selected from the group consisting of carnosine, anserine, and balenine. In the second embodiment, component (d) is one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof. That is, as with component (b) in the first embodiment, component (d) may contain one or more substances. That is, for example, component (d) may contain only uridine, only inosine, or uridine and inosine. For example, component (d) may contain uridine and / or one or more uridine precursors, and / or inosine and / or one or more inosine precursors.

[0019] Unless otherwise specified, the substances contained in the components of this embodiment (NAD precursor, uridine, uridine precursor, inosine, inosine precursor, β-alanine, and β-alanine precursor) also include their tautomers, geometric isomers (cis and trans isomers, E and Z isomers), optical isomers (D and L isomers), etc.

[0020] The composition according to the first embodiment has the effect of increasing the amount of intracellular ATP and NAD. Therefore, the composition according to the first embodiment can be used as a pharmaceutical composition for the prevention or treatment of diseases caused by a decrease in ATP or NAD in the body, including, but not limited to, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, spinocerebellar degeneration, dementia with Lewy bodies, and Huntington's disease; age-related diseases such as sarcopenia, frailty, and locomotive syndrome; and ischemic heart diseases such as myocardial infarction and angina pectoris. Furthermore, the composition according to the second embodiment has the effect of increasing muscle strength. Therefore, the composition according to the first embodiment and the composition according to the second embodiment can be used as a pharmaceutical composition for the prevention or treatment of diseases caused by a decrease in muscle strength, including, but not limited to, sarcopenia and frailty. Furthermore, the compositions according to the first embodiment and the second embodiment can be provided as, in addition to pharmaceutical compositions, for example, food and beverage compositions, cosmetic compositions, feed compositions, and hygiene compositions, but are not limited to these compositions.

[0021] The substance contained in the composition of this embodiment may be a pharmaceutically acceptable salt. Examples of pharmaceutically acceptable salts include, but are not limited to, alkali metal and alkaline earth metal salts such as lithium, sodium, potassium, magnesium, and calcium when the substance contains an acidic group; salts with amines such as ammonia, methylamine, dimethylamine, trimethylamine, dicyclohexylamine, tris(hydroxymethyl)aminomethane, N,N-bis(hydroxyethyl)piperazine, 2-amino-2-methyl-1-propanol, ethanolamine, N-methylglucamine, and L-glucamine; and salts with basic amino acids such as lysine, δ-hydroxylysine, and arginine. When the substance has a basic group, examples of the salt include salts with mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; salts with organic acids such as methanesulfonic acid, benzenesulfonic acid, paratoluenesulfonic acid, acetic acid, propionate, tartaric acid, fumaric acid, maleic acid, malic acid, oxalic acid, succinic acid, citric acid, benzoic acid, mandelic acid, cinnamic acid, lactic acid, glycolic acid, glucuronic acid, ascorbic acid, nicotinic acid, and salicylic acid; and salts with acidic amino acids such as aspartic acid and glutamic acid.

[0022] The dosage form of the composition according to this embodiment is not particularly limited and may be oral or parenteral, such as tablets, capsules, granules, powders, syrups, suspensions, suppositories, ointments, creams, gels, patches, inhalants, injections, and eye drops. These formulations are prepared according to conventional methods. Liquid formulations may be dissolved or suspended in water or other suitable solvents before use. Tablets and granules may also be coated by known methods. Injectable formulations are prepared by dissolving the active ingredient in water, but may also be dissolved in saline or glucose solution as needed, and buffers and preservatives may also be added. Furthermore, to promote absorption from areas with well-developed salvage pathways, such as the oral mucosa and intestinal mucosa, dosage forms that prolong the residence time in these areas are preferred, such as troches, lozenges, and enteric-coated formulations.

[0023] The type of formulation additive used in the production of the composition of this embodiment, the ratio of the formulation additive to the active ingredient (for example, in the first embodiment, component (a) and component (b)), the production method, etc. can be appropriately selected by those skilled in the art. As the formulation additive, inorganic or organic substances, or solid or liquid substances can be used, and generally, the formulation additive can be blended in an amount of 1 to 90% by weight based on the weight of the active ingredient. Specific examples of pharmaceutical additives include lactose, glucose, mannitol, dextrin, cyclodextrin, starch, sucrose, magnesium aluminometasilicate, synthetic aluminum silicate, sodium carboxymethylcellulose, hydroxypropyl starch, calcium carboxymethylcellulose, ion exchange resins, methylcellulose, gelatin, gum arabic, hydroxypropyl cellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, light anhydrous silicic acid, magnesium stearate, talc, tragacanth, bentonite, Veegum, titanium oxide, sorbitan fatty acid esters, sodium lauryl sulfate, glycerin, fatty acid glycerin esters, purified lanolin, glycerogelatin, polysorbate, macrogol, vegetable oils, wax, liquid paraffin, white petrolatum, fluorocarbons, nonionic surfactants, propylene glycol, and water.

[0024] To prepare solid formulations for oral administration, the active ingredient is mixed with excipients such as lactose, starch, crystalline cellulose, calcium lactate, and anhydrous silicic acid to form a powder, or, if necessary, with binders such as sucrose, hydroxypropyl cellulose, and polyvinylpyrrolidone, and disintegrants such as carboxymethylcellulose and calcium carboxymethylcellulose, followed by wet or dry granulation to form granules. To prepare tablets, these powders and granules may be compressed as is or with the addition of lubricants such as magnesium stearate and talc. These granules or tablets may be coated with enteric-coated bases such as hydroxypropylmethylcellulose phthalate and methacrylic acid-methyl methacrylate polymer to form enteric-coated formulations, or coated with ethylcellulose, carnauba wax, and hydrogenated oil to form sustained-release formulations. To prepare capsules, the powder or granules may be filled into hard capsules, or the active ingredient may be dissolved in glycerin, polyethylene glycol, sesame oil, olive oil, or the like and then coated with a gelatin membrane to form soft capsules.

[0025] To prepare an injection among parenteral preparations, the active ingredient can be dissolved in distilled water for injection, if necessary, together with a pH adjuster such as hydrochloric acid, sodium hydroxide, lactose, lactic acid, sodium, sodium monohydrogen phosphate, sodium dihydrogen phosphate, etc., and an isotonic agent such as sodium chloride or glucose, and then sterile filtered and filled into an ampule, or the solution can be further mixed with mannitol, dextrin, cyclodextrin, gelatin, etc., and then vacuum lyophilized to produce an injection that can be dissolved before use. Alternatively, the active ingredient can be emulsified in water with lecithin, polysorbate 80, polyoxyethylene hydrogenated castor oil, etc. to produce an emulsion for injection.

[0026] Eye drops may be prepared in the form of a liquid in which the active ingredient is suspended or dissolved in an aqueous solvent (for example, phosphate buffered saline).

[0027] Ointments and creams can be produced by kneading and mixing the active ingredient with a base and additives. Oil-based ointments can be produced, for example, by warming and melting an oil-based base such as oils, waxes, or hydrocarbons such as paraffin, adding the active ingredient, mixing to dissolve or disperse the active ingredient, and kneading until the entire mixture is homogeneous. Water-soluble ointments can be produced, for example, by warming and melting a water-soluble base such as macrogol, adding the active ingredient, and kneading until the entire mixture is homogeneous. Creams can be produced, for example, by adding the active ingredient to an oil phase containing petrolatum, higher alcohol, or the like, either as is or with the addition of an emulsifier or other additive, or to an aqueous phase containing purified water or with the addition of an emulsifier or other additive, heating each phase, and emulsifying the oil and aqueous phases by stirring until the entire mixture is homogeneous.

[0028] To prepare a parenteral rectal preparation, the active ingredient may be dissolved by wetting together with a suppository base such as cacao butter, tri-, di-, or monoglycerides of fatty acids, or polyethylene glycol, and then poured into a mold and cooled; alternatively, the active ingredient may be dissolved in polyethylene glycol, soybean oil, or the like, and then coated with a gelatin film.

[0029] The dosage and frequency of administration of the composition according to this embodiment are not particularly limited and can be appropriately selected at the discretion of a physician depending on factors such as the purpose of preventing and / or treating the condition to be improved or the progression of the disease being treated, the type of disease, and the patient's weight and age. Generally, the daily oral dose for adults is approximately 0.01 to 1000 mg (weight of active ingredient), and can be administered once a day or in divided doses several times a day, or every few days. When used as an injection, it is desirable to administer a daily dose of 0.001 to 100 mg (weight of active ingredient) to adults continuously or intermittently.

[0030] The composition ratio of the components contained in the composition according to this embodiment can be easily determined by a specialist in the relevant technical field (e.g., a physician, dentist, pharmacist, veterinarian, or other specialist familiar with health maintenance, etc.) depending on the purpose of administration, the health condition of the recipient, etc. Therefore, the dosage of each component and the component ratio in the composition are described below as an example, without any particular intention to limit them. First, the minimum amount of NAD precursor (e.g., NR or NMN) required for the composition according to the first embodiment to exert its ATP and NAD enhancing effect, when converted into a dosage per 1 kg of human body weight, is not particularly limited, but may be, for example, about 0.1 mg / kg. The daily dosage of each component is not particularly limited, but may be, for example, 0.1 mg to 3000 mg / day when the NAD precursor is NR, 0.1 mg to 3000 mg / day when the NAD precursor is NMN, 0.1 mg to 5000 mg / day for uridine (or uridine precursor), or 0.1 mg to 5000 mg / day for inosine (or inosine precursor). Furthermore, when a SIRT1 activator is administered in combination, the dosage is not particularly limited, but may be, for example, 1 mg to 1000 mg / day when the SIRT1 activator is resveratrol, or 0.1 to 1000 mg / day for other SIRT1 activators. Furthermore, the ratio of components contained in the composition according to the first embodiment is not particularly limited, and may be, for example, NR (or NMN):uridine (or uridine precursor) of about 0.001 to 1000, NR (or NMN):inosine (or inosine precursor) of about 0.001 to 1000, NR (or NMN):uridine (or uridine precursor):inosine (or inosine precursor) of about 0.001 to 1000, or NR (or NMN):uridine (or uridine precursor):inosine (or inosine precursor):SIRT1 activator of about 0.001 to 1000. Next, the minimum amount of β-alanine or a β-alanine precursor required for the composition of the second embodiment to exert its muscle-strengthening effect is not particularly limited, but may be, for example, about 0.1 g / kg to 0.2 g / kg for β-alanine, and about 0.01 g / kg to 0.1 g / kg for anserine (a β-alanine precursor).Furthermore, the ratio of the components contained in the composition according to the second embodiment is not particularly limited, and may be, for example, β-alanine (or β-alanine precursor):uridine (or uridine precursor) of about 100 to 0.100, β-alanine (or β-alanine precursor):inosine (or inosine precursor) of about 100 to 0.1, or β-alanine (or β-alanine precursor):uridine (or uridine precursor):inosine (or inosine precursor) of about 100 to 0.1.

[0031] The compositions of this embodiment may be formulated as sustained-release formulations, such as implants and microencapsulated delivery systems, using carriers that can prevent immediate elimination from the body. Such carriers include biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such materials can be readily prepared by those skilled in the art. Liposomal suspensions can also be used as pharmaceutically acceptable carriers. Liposomes can be prepared using a lipid composition containing, but not limited to, phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanol (PEG-PE), filtered through an appropriate pore size filter to obtain a suitable size, and purified by reverse-phase evaporation.

[0032] The composition according to this embodiment may be provided in the form of a kit together with instructions for administration, etc. For example, components (a) and (b) according to the first embodiment, and components (c) and (d) according to the second embodiment may each be stored in a separate container, and components (a) and (b) (the composition according to the first embodiment), or components (c) and (d) (the composition according to the second embodiment) may be combined and provided as a kit. The instructions to be provided with the kit may be printed on paper or the like, or may be stored on an electromagnetically readable medium such as a CD-ROM or DVD-ROM and provided to the user, or may be an electronic file available via the Internet, etc.

[0033] When the composition according to the present embodiment is provided as a food or beverage composition, its form is not particularly limited, and may be, for example, beverages such as soft drinks and nutritional drinks, confectioneries such as candy, gum, jelly, cream, and ice cream, dairy products such as milk drinks, fermented milk, drinkable yogurt, and butter, or other supplements (nutritional supplements). Furthermore, when the composition according to the present embodiment is provided as a cosmetic composition, its form is not particularly limited, and may be, for example, a cream, lotion, spray, or foam. The food or beverage composition and cosmetic composition according to the present embodiment are not particularly limited, and may be provided, for example, as anti-aging cosmetics or supplements for preventing or improving age-related skin wrinkles and dryness, or supplements for maintaining or increasing muscle strength.

[0034] When the composition according to the present embodiment is provided as a hygiene composition, its form is not particularly limited and may be, for example, toothpaste, skin cream, soap, shampoo, aerosol, mist, coating agent, etc. The composition according to the present embodiment may also be a feed composition or a pharmaceutical composition for animals that can be ingested by non-human animals. The animal feed referred to here includes not only so-called animal feeds that are ingested as a necessary nutritional source for animals, but also animal supplements that are given to animals as a luxury item.

[0035] Furthermore, the composition according to this embodiment may further contain an XOR inhibitor (febuxostat, NC-2500, allopurinol, etc.).

[0036] The third embodiment is a method for preventing or treating a disease caused by a decrease in ATP and / or NAD in the body, comprising administering to a subject the composition according to the first embodiment, i.e., a composition comprising a combination of the following (a) and (b): (a) an NAD precursor or a pharmaceutically acceptable salt thereof, (b) uridine, a uridine precursor or a pharmaceutically acceptable salt thereof, and / or inosine, an inosine precursor or a pharmaceutically acceptable salt thereof.

[0037] Furthermore, a fourth embodiment is a method for preventing or treating a disease caused by muscle weakness, characterized by administering the composition according to the second embodiment, i.e., the following (c) and (d) in combination: (c) β-alanine, a β-alanine precursor, or a pharmaceutically acceptable salt thereof, (d) uridine, a uridine precursor, or a pharmaceutically acceptable salt thereof, and / or inosine, an inosine precursor, or a pharmaceutically acceptable salt thereof.

[0038] Here, "treatment" means preventing or alleviating the progression or worsening of the pathological condition of the target disease, etc. Furthermore, "prevention" refers to a treatment aimed at preventing the onset of the target disease, etc. in advance. The subjects of the treatment method and prevention method according to this embodiment are not particularly limited, and may be any animal classified as a mammal, such as humans, pet animals such as dogs, cats, and rabbits, and livestock animals such as cows, pigs, sheep, and horses, in addition to humans. A particularly preferred "mammal" is humans.

[0039] When this specification is translated into English and includes the singular words "a," "an," and "the," it is intended to include not only the singular but also the plural, unless the context clearly indicates otherwise. Furthermore, in this specification, "about" or "approximately" refers to a numerical range of ±10%. The present invention will be further explained below with reference to examples, but these examples are merely illustrative of embodiments of the present invention and do not limit the scope of the present invention.

[0040] I. Enhancement of Intracellular ATP and NAD Levels I-1. Materials and Methods I-1-1. Reagents [1',2',3',4',5'-13C5]inosine and [1',2',3',4',5'-13C5]uridine were purchased from Omicron Biochemicals. 13 C5 D-glucose, 13 C5 D-ribose, 15N4 inosine was purchased from Cambridge Isotope Laboratories. Uridine and guanosine were purchased from Tokyo Chemical Industry Co., Ltd. Inosine, hypoxanthine, and nicotinamide riboside were purchased from Sigma-Aldrich. Orotidine was obtained from Santa Cruz Biotechnology. All other reagents were purchased from Fujifilm Wako Pure Chemical Industries, Ltd.

[0041] I-1-2. Cell Culture HEK293 (RCB1637) cells were obtained from the RIKEN BioResource Research Center (Tsukuba, Japan). Growth medium was D-MEM (0.45% glucose) supplemented with 10% FBS and 1% penicillin / streptomycin / amphotericin, and was changed every two days. Cells were cultured until 80% confluent before experiments. Experimental medium was D-MEM (0.45% glucose or no glucose) containing 10% FBS that had been dialyzed to remove inosine and hypoxanthine. When compounds were added to the experimental medium, the same volume of solvent was also added to the control group.

[0042] I-1-3. Extraction of Intracellular Metabolites for HPLC Extraction of intracellular metabolites was performed according to a previously published method (Sato et al., Gout and Uric & Nucleic Acids, 47:157-168 2023). Briefly, cells were washed three times with ice-cold phosphate-buffered saline (PBS) and incubated on ice for 30 minutes with 5% perchloric acid containing 50 μM caffeine as an internal standard. The supernatant was neutralized with 3 M potassium carbonate and centrifuged to remove the precipitate. Finally, the supernatant was equilibrated with 0.1 M potassium phosphate buffer, the HPLC elution buffer.

[0043] I-1-4. Extraction of intracellular metabolites for MS analysis. Cells were washed three times with ice-cold 150 mM ammonium formate. After removing the wash solution, ice-cold methanol was poured into the wells and gently swirled for 30 seconds to cover the surface. An equal volume of water as methanol was added to the wells, mixed thoroughly, and then allowed to stand on ice for 30 minutes. The supernatant was collected and centrifuged to remove cellular debris.

[0044] I-1-5. Protein Concentration Measurement After washing with ice-cold PBS, cell lysis buffer (50 mM Tris-HCl pH 7.8, 0.25 M sucrose, 0.3 mM EDTA, protease inhibitor cocktail) was added. After 30 minutes, the lysate was transferred to a microtube and centrifuged to remove precipitate. Protein concentration in the lysate was measured using the Pierce 660 nm Protein Assay (Thermo Fisher Scientific). All measurements were performed in duplicate.

[0045] I-1-6. Quantitation of nucleotides, nucleosides, and bases by HPLC HPLC analysis was performed using an LC-20AD (Shimadzu Corporation) and a SUPELCOSIL LC-18-T (25 cm × 4.6 mm, 5 μm) column mounted on a Supelguard TM The analysis was performed using an LC-18 (Sigma-Aldrich). A Shimadzu SPD-M20A photodiode array detector was used. The elution method was based on the column supplier's report, with some modifications. For nucleotides, the mobile phase consisted of (A) 0.03 M potassium phosphate buffer (pH 7.5), 10 mM tetrabutylammonium hydrogen sulfate, and (B) methanol. Elution was performed at 40°C with a flow rate of 0.7 mL / min. The injection volume of each sample was 30 μL. The absorbance of caffeine, UTP, and other compounds was measured at 273 nm, 268 nm, and 254 nm, respectively, and peaks were detected. The concentration of each compound in the sample was calculated from the ratio of its peak area to that of the internal standard (caffeine).

[0046] I-1-7. HPLC-MS / MS analysis of stable isotopes Intracellular ATP, UTP, and NAD were analyzed using an AKTA purification system (GE Healthcare) connected to an InertSustain AX-C18 metal-free column (3 μm, 4.6 mm × 150 mm, GL Science). + The mobile phase consisted of (A) 10 mM ammonium formate and (B) 50% methanol, 100 mM ammonium formate. The flow rate was 0.7 mL / min. ATP, UTP, and NAD +Each fraction was subjected to ESI-QTOF-MS (impact II, Bruker). The data were analyzed using the accompanying software, DataAnalysis (Bruker).

[0047] Statistical analysis was performed using R and R studio version 4.2.1 (R Foundation) using Student's t-test for comparisons between two groups and Tukey's HSD test for comparisons between multiple groups. All data are expressed as mean values, and error bars indicate standard deviations. Statistical differences were considered p ≤ 0.05.

[0048] I-2. Results I-2-1. Quantification of intracellular nucleotides I-2-1-1. Intracellular ATP and NAD levels by various nucleosides + The effect on the amount of each nucleotide in HEK293 cells cultured for 6 hours in a test medium containing glucose-free medium to which each compound was added at a final concentration of 100 μM is shown in Figure 1. ATP significantly increased in the inosine and adenosine groups, and slight but significant increases were also observed in the uridine, cytidine, NR, and orotidine groups. NAD + was significantly increased only in the NR group, whereas it was significantly decreased in the adenosine group.

[0049] I-2-1-2. Intracellular ATP and NAD + Figure 2 shows the additive or synergistic effects of the combined use of inosine, NR, and / or uridine on the amount of each nucleotide in HEK293 cells cultured for 6 hours in test medium containing glucose-free medium to which each compound was added at a final concentration of 100 μM. ATP showed a similar increase in all groups to which inosine was added. A significant increase in ATP was also observed in the uridine / NR combination group without inosine. NAD + was significantly increased in all groups with NR, and the combination of NR with inosine or uridine significantly increased NAD levels compared with NR alone. + The highest NAD levels were observed in the inosine and NR combination group and the inosine, uridine and NR combination group. + In addition, even in the inosine / uridine combination group without NR, NAD +was significantly increased.

[0050] I-2-1-3. Comparison of inosine and hypoxanthine in the effect of enhancing intracellular ATP levels. Figure 3 shows the levels of each nucleotide in HEK293 cells cultured for 1 or 6 hours in glucose-free medium containing each compound added to a final concentration of 100 μM. Intracellular ATP levels increased significantly in all groups compared to the control group. The inosine group treated for 1 hour showed significantly higher levels than the hypoxanthine group. No difference was observed between the hypoxanthine and ribose groups and the hypoxanthine group.

[0051] I-2-1-4. Intracellular NAD + NR and other NADs in relation to their quantity-enhancing effects + Comparison of precursors Figure 4 shows the amount of NAD in HEK293 cells cultured for 3 hours in a test medium containing 0.1% glucose and containing each compound at a final concentration of 100 μM. + Precursors significantly increase NAD + The NR-added group showed the highest intracellular NAD + The quantity was shown.

[0052] I-2-2. Stable isotope tracer experiments I-2-2-1. Intracellular ATP and NAD + Figure 5 shows the results of MS measurements of each nucleotide in HEK293 cells cultured for a certain period of time in a test medium containing glucose-free medium to which a stable isotope had been added (final concentrations are shown in the graph). Molecules that incorporate phosphoribosyl pyrophosphoric acid (PRPP), synthesized from stable isotope glucose or ribose, appear as peaks with an m / z shift of +5 from the monoisotopic mass (Figure 5A. For ATP, the m / z shifts from 506 to 511). ATP and NAD +The intensity of the peak with a m / z shift of +5 is shown over time (Fig. 5B, C). Comparing over 6 hours, stable isotope PRPP was incorporated into 20.8% of ATP at 1 mM glucose, but only 4.8% at 100 μM glucose and 0.9% at 100 μM ribose. NAD + 16.2% of ATP and NAD were labeled at 1 mM glucose for 6 hours. + In both cases, the uptake rate of ribose was significantly lower than that of glucose. Ribose is involved in PRPP synthesis and the production of ATP and NAD. + It was suggested that it did not contribute much to the synthesis of

[0053] I-2-2-2. Intracellular ATP and NAD + Incorporation of inosine-derived ribose into ATP and NAD in HEK293 cells cultured for a certain period of time in glucose-free medium or high glucose (0.45%) medium supplemented with a stable isotope at a final concentration of 100 μM. + The MS measurement results are shown in Figure 6. ATP and NAD + The intensity of the peak shifted by +5 m / z is shown over time (Fig. 6B, C). There is no enzyme that directly phosphorylates inosine to produce a nucleotide. 13 C5 inosine induces ATP and NAD at m / z +5 + The increase in NAD indicated that ribose monophosphate, produced by the hydrolysis of inosine, becomes PRPP and is then used for nucleotide synthesis. The incorporation rate of 100 μM inosine into ATP was 22.6% under glucose-free conditions for 6 hours, which is approximately five times the incorporation rate of glucose at the same concentration (4.8%, Figure 5B). This suggests that inosine-derived ribose is efficiently used for ATP synthesis. Furthermore, the incorporation rate under high glucose conditions was significantly lower than under glucose-free conditions, suggesting that inosine-derived ribose and glucose compete for PRPP precursors. + The same tendency as for ATP was observed for its uptake into the blood.

[0054] I-2-2-3. Intracellular ATP and NAD+ Incorporation of uridine-derived ribose into ATP and NAD in HEK293 cells cultured for a certain period in glucose-free medium or high glucose (0.45%) medium supplemented with a stable isotope at a final concentration of 100 μM. + The MS measurement results are shown in Figure 7. ATP and NAD + The ratio of the intensity of the peak with a m / z shift of +5 is shown over time (Fig. 7B, C). 13 C5 uridine binds to ATP and NAD + The only pathway that labels uridine is ribose monophosphate, which is generated by the hydrolysis of uridine with uridine perphosphate, and is used for nucleotide synthesis via PRPP. The incorporation rate of 100 μM uridine into ATP was 10.7% under glucose-free conditions for 6 hours, suggesting that uridine is used for ATP synthesis more efficiently than glucose. Furthermore, under high glucose conditions, the incorporation rate was significantly lower than under glucose-free conditions, suggesting that uridine-derived ribose and glucose compete for PRPP precursors. NAD + The same tendency as for ATP was observed for its uptake into the blood.

[0055] I-2-2-4. Inhibition of uridine-derived ribose incorporation into intracellular ATP by uridine phosphorylase inhibitors. 5-benzyl acyclouridine (BAU), an inhibitor of uridine phosphorylase, an enzyme that catalyzes the hydrolysis of uridine, 13 The effect of C5 uridine on tracer experiments was investigated. The results of MS measurements of ATP in HEK293 cells cultured for 6 hours in a glucose-free medium containing a stable isotope at a final concentration of 100 μM are shown. The percentage of the intensity of the peak with a m / z shift of +5 is shown (Figure 8). Inhibition of uridine phosphorylase resulted in: 13 The incorporation rate of C5 uridine into ATP was significantly reduced (7.3% to 1.1%), and its incorporation into ATP was almost completely inhibited, indicating that ribose monophosphate generated by uridine hyperphosphorylation was utilized for ATP synthesis via PRPP.

[0056] II. Enhancement of muscle endurance by combined administration of β-alanine and inosine II-1. Methods Seven-week-old male C57BL6j mice (CLEA Japan) were randomly divided into a control group and a treatment group. The treatment group received free access to tap water containing 1.2% (w / v) β-alanine (Fujifilm Wako Pure Chemical Industries, Ltd.) for 9 days or 2 weeks. On the day of the treadmill running endurance test, the mice were given NaHCO 3 dissolved in tap water one hour before running. 3 Mice were orally administered 500 mg / kg of NaHCO3 and 165 mg / kg of MgO. NaHCO3 was administered to increase blood and renal pH, while MgO was administered to convert to MgCl2 in the stomach and absorb unwanted CO2 gas produced in the stomach during NaHCO3 administration (Ando et al., Biol. Pharm. Bull. 44, 844-852, 2021). Additionally, 40 min before running, mice were orally administered a single 25 mg / kg dose of inosine (Sigma-Aldrich) dissolved in distilled water. The control group received distilled water instead of β-alanine and inosine, but otherwise received the same treatment as the treatment groups. To assess muscular endurance, mice were run on a treadmill (Melquest Co., Ltd.) and the running time was measured. The treadmill was set at a 0% incline, and running speed was gradually increased every 2 min until reaching 20 m / min. The mice were forced to run using electrical stimulation at 0.4 mA per stimulation, and the mice were terminated from the fatigue limit when the number of electrical stimulations reached 130.

[0057] II-2. Results Figure 9 shows the results of 9 consecutive days of β-alanine administration followed by a single dose of inosine, and Figure 10 shows the results of 2 consecutive weeks of β-alanine administration followed by a single dose of inosine. Mice were given β-alanine (1.2% w / v) in drinking water for 9 consecutive days, then given a single dose of inosine (25 mg / kg), and then subjected to treadmill running. Running time was extended, demonstrating enhanced muscle endurance. On the other hand, mice given β-alanine (1.2% w / v) in drinking water for 2 consecutive weeks followed by a single dose of inosine (25 mg / kg) did not extend running time, and no enhancement of muscle endurance was observed. These results suggest that the combination of short-term continuous administration of β-alanine (within 2 weeks) and a single dose of inosine enhances muscle endurance.

[0058] The present invention is expected to be used in the fields of medicine as well as food and beverages.

Claims

1. A composition for increasing the amount of intracellular adenosine triphosphate (ATP) and intracellular nicotinamide adenine dinucleotide (NAD), comprising a combination of the following: (a) an NAD precursor, or a pharmaceutically acceptable salt thereof, and (b) one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof.

2. The composition of claim 1, wherein the NAD precursor is nicotinamide mononucleotide (NMN) and / or nicotinamide riboside (NR).

3. The composition according to claim 1 or 2, wherein (b) is one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof.

4. The composition of claim 1 or 2, wherein (b) is one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof.

5. The composition of claim 1 or 2, further comprising a SIRT1 activator.

6. The composition according to claim 5, wherein the SIRT1 activator is a citrus peel extract.

7. The composition according to claim 6, wherein the citrus fruit is calamandarin, shikuwasa and / or shiranui.

8. The composition of claim 5, wherein the SIRT1 activator is 3',4',3,5,6,7,8-heptamethoxyflavone, 5-demethylnobiletin, and / or tangeretin.

9. The composition according to claim 1 or 2, which is a pharmaceutical composition, a food or beverage composition, or a cosmetic composition.

10. The composition according to claim 5, which is a pharmaceutical composition, a food or beverage composition, or a cosmetic composition.

11. A composition for increasing muscle strength, comprising a combination of the following (c) and (d): (c) one or more substances selected from the group consisting of β-alanine and β-alanine precursors, or pharmaceutically acceptable salts thereof, and (d) one or more substances selected from the group consisting of uridine and uridine precursors, or pharmaceutically acceptable salts thereof, and / or one or more substances selected from the group consisting of inosine and inosine precursors, or pharmaceutically acceptable salts thereof.

12. The composition of claim 11, wherein the beta-alanine precursor is one or more selected from the group consisting of carnosine, anserine, and balenine.

13. The composition according to claim 11 or 12, which is a pharmaceutical composition, a food or beverage composition, or a cosmetic composition.

Images