Combination of Urothyrin and Ketone Bodies

Abstract

The present invention relates to methods involving the administration of a combination of a urolithin and one or more ketone bodies, which result in the provision of beneficial health effects, such as improved resistance to inflammation, mitochondrial function, and cellular metabolism. The methods are useful, for example, for improving the health and well-being of a subject, particularly an elderly or infirm individual, as well as for improving the fitness, muscle performance, and / or endurance of an exerciser. The methods are also useful for treating or preventing various conditions, such as conditions associated with insufficient mitochondrial activity, excessive inflammation, and / or muscle-related disorders. The present invention relates to compositions comprising a urolithin and one or more ketone bodies.

Description

[Technical Field]

[0001] The present invention relates to methods involving the administration of a combination of a urolithin and one or more ketone bodies, which result in the provision of beneficial health effects, such as improved resistance to inflammation, mitochondrial function, and cellular metabolism. The methods are useful, for example, for improving the health and well-being of a subject, particularly an elderly or infirm individual, as well as for improving the fitness, muscle performance, and / or endurance of individuals engaging in physical activity. The methods are also useful for treating or preventing various conditions, such as conditions associated with insufficient mitochondrial activity, excessive inflammation, and / or muscle-related disorders. The present invention relates to compositions comprising a urolithin and one or more ketone bodies. [Background technology]

[0002] Urolithins have been proposed as treatments for various conditions associated with insufficient mitochondrial activity, including obesity, reduced metabolic rate, metabolic syndrome, diabetes, cardiovascular disease, hyperlipidemia, neurodegenerative diseases, cognitive disorders, mood disorders, stress, and anxiety disorders, for weight management, or to increase muscle or mental performance. See U.S. Patent No. 5,623,999 (Amazentis SA). The use of urolithins for the treatment of various neoplastic diseases is described in U.S. Patent No. 5,623,999 (The Regents of the University of California).

[0003] Patent document 3 (derived from International Application No. PCT / US2013 / 48310) discloses a method for increasing autophagy (specifically including mitosis) in a cell, comprising contacting a cell with an effective amount of a urolithin or a pharmaceutically acceptable salt thereof, thereby increasing autophagy (specifically including mitosis) in the cell. Administration may be to a subject having a disease or condition selected from metabolic stress, cardiovascular disease, endothelial cell dysfunction, sarcopenia, muscle degenerative disease, Duchenne muscular dystrophy, alcoholic liver disease, non-alcoholic fatty liver disease, drug-induced liver or muscle damage, alpha 1-antitrypsin deficiency, ischemia / reperfusion injury, inflammation, skin aging, inflammatory bowel disease, Crohn's disease, obesity, metabolic syndrome, type II diabetes, hyperlipidemia, osteoarthritis, neurodegenerative disease, Alzheimer's disease, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis, age-related macular degeneration, mitochondrial disease (including, for example, poor growth, loss of muscle coordination, muscle weakness, visual impairment, hearing impairment, heart disease, liver disease, kidney disease, gastrointestinal disorders, respiratory disorders, neurological disorders, autonomic dysfunction, and sometimes learning disabilities, and dementia (as a result of mitochondrial disease)), muscle disease, cancer, cognitive disorders, stress, and mood disorders.

[0004] In particular, urolithins have been proposed as therapeutic agents for muscle-related conditions. Muscle-related conditions include myopathy and neuromuscular diseases. Examples of such conditions include Duchenne muscular dystrophy, acute sarcopenia, such as muscle atrophy and / or cachexia, for example, those associated with burns, bed rest, limb immobilization, or major chest, abdominal, neck, and / or orthopedic surgery, amyotrophic lateral sclerosis, and multiple sclerosis. Age-related muscle loss is a particularly prevalent condition. Cachexia due to prolonged immobilization or other diseases, such as cancer, is another condition often characterized by reduced muscle performance.

[0005] Another biological function of urolithin A is the attenuation of harmful inflammatory responses. This is of particular clinical significance because aging and most age-related diseases are associated with chronic, low-grade inflammation. This process, recently termed inflamm-aging, contributes to the age-related decline in cellular and organismal function.

[0006] Effective muscle function and physical performance are important for healthy individuals as well as individuals suffering from diseases, especially the elderly, to have a high quality of life at all ages. Improving muscle performance is of particular interest to athletes. For example, increasing muscle contraction strength, increasing muscle contraction amplitude, or shortening muscle reaction time between stimulation and contraction are all beneficial to individuals, especially athletes. For elderly individuals who suffer from age-related muscle function decline, including muscle loss / wasting, or individuals suffering from muscle wasting due to cachexia, improving muscle and physical performance is important for fundamental aspects of daily function, such as walking speed and distance that can be walked unassisted.

[0007] Ketone bodies are a group of carbonyl-containing compounds, including acetoacetate, beta-hydroxybutyrate (BHB), and acetone, that are produced by the liver from fatty acids under certain conditions. They are known to induce mitochondrial biogenesis and have been suggested to be beneficial for muscle performance, metabolism, neuroprotection, healthy aging, cardiovascular health, cognitive performance, and mental health. Ketone bodies have also been suggested to be useful in the treatment of cancer. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Patent Publication No. 2012 / 088519 [Patent Document 2] International Patent Publication No. 2007 / 127263 [Patent Document 3] International Patent Publication No. 2014 / 004902 Summary of the Invention

[0009] The present invention provides combinations comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0010] The invention provides methods for providing a beneficial health effect to a subject, the methods comprising administering to the subject a combination comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0011] The present invention provides combinations comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) for providing a beneficial health effect to a subject.

[0012] The present invention provides for the use of a combination comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) to provide beneficial health effects in a subject.

[0013] The invention provides use of a combination comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) in the manufacture of a medicament for providing a beneficial health effect in a subject.

[0014] The invention provides methods of treatment that involve the administration of a combination comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0015] The present invention provides a combination comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) for use as a medicament.

[0016] The present invention provides for the use of a combination comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0017] The present invention provides compositions comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0018] The invention provides methods for providing a beneficial health effect to a subject, the methods comprising administering to the subject a composition comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0019] The present invention provides compositions comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) for providing a beneficial health effect to a subject.

[0020] The present invention provides uses of compositions comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) to provide a beneficial health effect in a subject.

[0021] The invention provides use of a composition comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) in the manufacture of a medicament for providing a beneficial health effect in a subject.

[0022] The invention provides methods of treatment that include administering a composition comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0023] The present invention provides compositions comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) for use as a medicament.

[0024] The invention provides use of a composition comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof) in the manufacture of a medicament.

[0025] The invention provides kits comprising a composition comprising a ketone body (or a salt, prodrug, metabolite, or derivative thereof) together with a separate composition comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof). [Brief explanation of the drawings]

[0026] [Figure 1] Graph showing the effect of urolithin A pretreatment on IL-6 secretion [Figure 2] Graph showing the effect of BHB, UA, and their combination on IL-6 secretion DETAILED DESCRIPTION OF THE INVENTION

[0027] The present invention provides combinations comprising or consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0028] Ketone bodies Ketone bodies are a group of carbonyl-containing compounds, including acetoacetate, beta-hydroxybutyrate, and acetone, that are produced by the liver from fatty acids under certain conditions. Ketone bodies are produced when fatty acid levels are elevated in the body and are metabolized for energy.

[0029] Preferably, the ketone body for use in the present invention is selected from acetoacetic acid, a salt, prodrug, metabolite, or derivative thereof, β-hydroxybutyric acid, a salt, prodrug, metabolite, or derivative thereof (e.g., (R)-β-hydroxybutyric acid, a salt, prodrug, metabolite, or derivative thereof, or (S)-β-hydroxybutyric acid, a salt, prodrug, metabolite, or derivative thereof), and acetone, a salt, prodrug, metabolite, or derivative thereof, or a mixture thereof. The ketone body is selected from β-ketopentanoic acid and β-hydroxypentanoic acid, a salt, prodrug, metabolite, or derivative thereof.

[0030] In some combinations and compositions of the present invention, the ketone body is acetoacetic acid, or a salt, prodrug, metabolite, or derivative thereof, or a mixture thereof. In some embodiments of the present invention, the ketone body is β-hydroxybutyric acid, or a salt, prodrug, metabolite, or derivative thereof, or a mixture thereof. In some embodiments of the present invention, the ketone body is acetone, or a salt, prodrug, metabolite, or derivative thereof, or a mixture thereof.

[0031] In some combinations and compositions of the present invention, the ketone body is acetoacetic acid. In some combinations and compositions of the present invention, the ketone body is beta-hydroxybutyric acid. In some combinations and compositions of the present invention, the ketone body is acetone.

[0032] In some embodiments of the invention, the combination or composition may include two or more ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof). In some embodiments of the invention, the combination and composition include acetoacetic acid and β-hydroxybutyric acid. In some embodiments of the invention, the combination and composition include acetoacetic acid and acetone. In some embodiments of the invention, the combination and composition include β-hydroxybutyric acid and acetone. In some embodiments of the invention, the combination and composition include acetoacetic acid, β-hydroxybutyric acid, and acetone.

[0033] The present invention also encompasses the use of suitable salts of ketone bodies, such as pharmaceutically acceptable salts. Suitable salts according to the present invention include those formed with organic or inorganic bases. Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts, such as potassium and sodium salts, alkaline earth metal salts, such as calcium and magnesium salts, and salts with organic bases, such as dicyclohexylamine, N-methyl-D-glucamine, morpholine, thiomorpholine, piperidine, pyrrolidine, mono-, di-, or tri-lower alkylamines, such as ethyl, tert-butyl, diethyl, diisopropyl, triethyl, tributyl, or dimethylpropylamine, or mono-, di-, or trihydroxy-lower alkylamines, such as mono-, di-, or triethanolamine.

[0034] The present invention also encompasses the use of prodrugs and derivatives of ketone bodies. Ketone body derivatives or ketone body prodrugs are compounds or species that can be converted or metabolized to ketone bodies. Suitable ketone body derivatives and ketone body prodrugs include, for example, ketone body esters (e.g., [(3R)-3-hydroxybutyl](3S)-3-hydroxybutanoate, [(3S)-3-hydroxybutyl](3R)-3-hydroxybutanoate, [(3S)-3-hydroxybutyl](3S)-3-hydroxybutanoate, or butanediol, particularly butane-1,3-diol, altrose, arabinose, dextrose, erythrose, fructose, galactose, glucose, glycerol, etc.). Examples of ketone body derivatives and ketone body prodrugs include esters formed with (S)-1,2-propanediol (e.g., esters formed with (S)-1,2-propanediol ...

[0035] The present invention also encompasses the use of metabolites of ketone bodies.

[0036] Those skilled in the art of organic chemistry will understand that many organic compounds can form complexes with solvents with which they react or from which they are precipitated or crystallized. These complexes are known as "solvates." Those skilled in the art will understand that the present invention also encompasses solvates of ketone bodies, as well as solvates of their salts, prodrugs, metabolites, and derivatives. Solvates include those in which the associated solvent is pharmaceutically acceptable. A hydrate (in which the associated solvent is water) is one example of a solvate.

[0037] Urolithin Urolithins are metabolites produced by the action of the intestinal microflora of mammals, including humans, on ellagitannins and ellagic acid. Ellagitannins and ellagic acid are compounds commonly found in foods such as pomegranates, nuts, and berries. Ellagitannins themselves are minimally absorbed in the intestine. Urolithins are a class of compounds having the representative formula (I) shown below. The structures of some particularly common urolithins are set forth in the table below with reference to formula (I). [ka] A, B, C, D, W, X, Y and Z are each independently selected from H and OH. [Table 1]

[0038] In practice, it is convenient to synthesize urolithins for commercial-scale production, and synthetic routes are described, for example, in U.S. Patent No. 5,623,999 and International Patent Publication No. 2019 / 0168972.

[0039] Particularly suitable compounds for use in the present invention are naturally occurring urolithins. Thus, Z is preferably OH, and W, X, and Y are preferably all H. When W, X, and Y are all H, A and B are both H, and C, D, and Z are all OH, the compound is urolithin C. When W, X, and Y are all H, A, B, and C are all H, and D and Z are both OH, the compound is urolithin A. Preferably, the urolithin used in the formulations of the present invention is urolithin A, urolithin B, urolithin C, and / or urolithin D. Most preferably, the urolithin used in the formulations of the present invention is urolithin A or urolithin B. Most preferably, the urolithin used in the formulations of the present invention is urolithin A. [ka]

[0040] Also referred to are isourolitins A and B. When W, X, and Y are all H, A, B, and D are all H, and C and Z are both OH, the compound is iso-urolithin A. When W, X, Y, and Z are all H, A, B, and D are all H, and C is OH, the compound is iso-urolithin B.

[0041] The present invention also encompasses the use of suitable salts of urolithins, e.g., pharmaceutically acceptable salts. Suitable salts according to the present invention include those formed with organic or inorganic bases. Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts, e.g., potassium and sodium salts, alkaline earth metal salts, e.g., calcium and magnesium salts, and salts with organic bases, e.g., dicyclohexylamine, N-methyl-D-glucamine, morpholine, thiomorpholine, piperidine, pyrrolidine, mono-, di-, or tri-lower alkylamines, e.g., ethyl-, tert-butyl-, diethyl-, diisopropyl-, triethyl-, tributyl-, or dimethyl-propylamine, or mono-, di-, or trihydroxy lower alkylamines, e.g., mono-, di-, or triethanolamine.

[0042] The present invention also encompasses the use of prodrugs of urolithins, such as amino acid derivatives of urolithins. One preferred group of prodrugs of urolithins for use in the present invention are compounds of formula (II): [ka] wherein A, B, C, D, W, X, Y, and Z are each independently H and OR 1 with the proviso that at least one of A, B, C, D, W, X, Y, and Z is selected from the group consisting of: 1 and Each R 1 are independently H or C(=O)R 2 and at least one R 1 The group is C(=O)R 2 and Each R2 but, CHR 3 NHR 4 Selected from R 4 is H and R 3 is a group selected from CH3, CH2CH(CH3)2, CH(CH3)CH2CH3, CH2Ph, CH2-3-(1H-indole), CH2CH2SCH3, CH2OH, CHOHCH3, CH2SH, CH2SeH and CH2PhpOH, The R 3 The group is optionally halogen, cyano, nitro, or OR A Or C 1- C4 alkyl, Or R 3 and R 4 together with the C and N atoms to which they are attached form a 5-membered heteroalkyl ring, The heteroalkyl ring may optionally be selected from the group consisting of halogen, cyano, nitro, OR A or C 1- C alkyl, R A optionally substituted with one or more halogen, cyano, or nitro groups; 1- C4 alkyl. Compounds of formula (II) are disclosed in International Patent Publication No. 2015 / 097231.

[0043] In one implementation of the invention, the urolithin is administered as a metabolite of the urolithin, such as a glucuronide or sulfate. Preferred metabolites are urolithin A 3-O-glucuronide, urolithin A 3-O-sulfate, urolithin B 3-O-glucuronide, and urolithin B 3-O-sulfate.

[0044] Those skilled in the art of organic chemistry will understand that many organic compounds can form complexes with solvents with which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates." Those skilled in the art will understand that the present invention also encompasses solvates of urolithins, as well as solvates of their salts, salts, prodrugs, metabolites, and derivatives. Solvates include those in which the associated solvent is pharmaceutically acceptable. A hydrate (in which the associated solvent is water) is one example of a solvate.

[0045] Preferably, urolithin (or its salt, prodrug, metabolite, or derivative) for use in the compositions of the invention is micronized. It has been found that micronized urolithin can be dissolved or suspended more rapidly and effectively than non-micronized urolithin. Micronized urolithin (or its salt, prodrug, metabolite, or derivative) preferably has a D of less than 100 μm. 50 More preferably, the urolithin (or salt, prodrug, metabolite, or derivative thereof) has a particle size of less than 75 μm, e.g., less than 50 μm, e.g., less than 25 μm, e.g., less than 20 μm, e.g., less than 10 μm. 50 More preferably, the urolithin (or salt, prodrug, metabolite, or derivative thereof) has a D in the range of 0.5 to 50 μm, for example, 0.5 to 20 μm, for example, 0.5 μm to 10 μm, for example, 1 μm to 10 μm. 50 Preferably, the urolithin (or salt, prodrug, metabolite, or derivative thereof) has a D of less than 100 μm. 90 More preferably, the urolithin (or salt, prodrug, metabolite, or derivative thereof) has a particle size of less than 75 μm, e.g., less than 50 μm, e.g., less than 25 μm, e.g., less than 20 μm, e.g., less than 15 μm. 90The urolithin (or salt, prodrug, metabolite, or derivative thereof) preferably has a D in the range of 5 μm to 100 μm, e.g., 5 μm to 50 μm, e.g., 5 to 20 μm. 90 Micronization can be achieved by methods established in the art, for example, compression milling, hammer milling, universal or pin milling, or jet milling (e.g., spiral jet milling or fluidized bed jet milling). Jet milling is particularly preferred.

[0046] In some embodiments, the present invention is directed to a method for providing a beneficial health effect to a subject, the method comprising administering to the subject a combination or composition of the present invention.

[0047] In some embodiments, the present invention is directed to a combination or composition of the present invention for providing a beneficial health effect to a subject.

[0048] In some embodiments, the present invention is directed to the use of a combination or composition of the present invention to provide a beneficial health effect to a subject.

[0049] In some embodiments, the present invention is directed to the use of a combination or composition of the present invention in the manufacture of a medicament for providing a beneficial health effect to a subject.

[0050] In some embodiments, the present invention is directed to a method of treatment comprising administering a combination or composition of the present invention to a subject.

[0051] In some embodiments, the present invention is directed to a combination or composition of the present invention for use as a medicament.

[0052] In some embodiments, the present invention is directed to the use of a combination or composition of the present invention for the manufacture of a medicament.

[0053] In some embodiments, the methods, agents, or combinations of the present invention reduce muscle glycogen and / or protein breakdown during or after exercise, aiding muscle recovery. In some embodiments, the subject is a healthy individual. In other embodiments, the subject has a muscle-wasting condition.

[0054] In some embodiments, the methods, medicaments, or combinations of the present invention reduce muscle wasting in a subject susceptible to muscle wasting. In some embodiments, the subject is a healthy individual. In other embodiments, the subject has a muscle wasting condition.

[0055] In some embodiments, the degree of reduction in protein breakdown during exercise, muscle recovery after exercise, or muscle wasting can be determined by the loss of leucine, isoleucine, and valine in a subject, which in some embodiments is reduced by at least 10% after administration of a ketone body ester compared to the loss of leucine, isoleucine, and valine in a subject not administered a ketone body or ketone body ester.

[0056] In some embodiments, the reduction in muscle glycogen breakdown during exercise can be determined by glycogen loss in a subject, which in some embodiments is reduced by at least 10% after administration of ketone bodies or ketone body esters compared to glycogen loss in a subject not administered ketone bodies or ketone body esters.

[0057] Alternatively, the reduction in muscle glycogen and / or protein breakdown during exercise can be determined by the rate of glycogen or protein loss, hi some embodiments, the rate of glycogen or protein loss during exercise is reduced compared to glycogen levels without administration of ketone bodies or ketone body esters.

[0058] In some embodiments, the methods, agents or combinations of the present invention reduce glycogen and / or protein breakdown in a subject.

[0059] In some embodiments, the methods, agents, or combinations of the present invention reduce muscle breakdown or slow muscle wasting in a subject susceptible to muscle breakdown or muscle wasting, hi some embodiments, the reduction in muscle breakdown is at least 50% less than in the same subject under comparable conditions in the absence of the methods, agents, or combinations of the present invention.

[0060] In some embodiments, the methods, agents, or combinations of the present invention maintain or improve muscle output in a subject. In some embodiments, the methods, agents, or combinations of the present invention maintain or improve skeletal muscle output. In some embodiments, the methods, agents, or combinations of the present invention maintain or improve output in other muscle types, such as cardiac muscle.

[0061] In some embodiments, the methods, medicaments or combinations of the present invention maintain or improve muscle power output in a subject, and the subject is a healthy individual.

[0062] In some embodiments, the methods, agents or combinations of the invention improve muscle power output in a subject, wherein the improved power output is at least 0.25% compared to placebo as measured in a controlled test over a 30 minute period.

[0063] In some embodiments, the methods, agents or combinations of the present invention improve muscle power output in a subject, wherein the increased power output is at least 1 watt relative to placebo as measured in a controlled test over a 30 minute period.

[0064] In some embodiments, the methods, medicaments, or combinations of the present invention find use in improving muscle performance, maintaining or improving muscle function, preventing a decline in muscle function, increasing muscle mass, and / or reducing muscle wasting. Improving muscle performance, improving or maintaining muscle function, increasing muscle mass, and / or reducing muscle wasting may be part of a medical procedure or may be for personal preference ("lifestyle") or cosmetic reasons. The compositions of the present invention also find use in reducing muscle inflammation after exercise and / or promoting muscle recovery after exercise.

[0065] The method, medicament or combination of the present invention may be for use as a medicament, as a dietary supplement, as a functional food, as a functional beverage, or as a medical food.

[0066] The method, agent, or combination of the present invention is useful in promoting muscle performance. Thus, the present invention provides the method, agent, or combination of the present invention for use in promoting muscle performance. The present invention also provides a method for promoting muscle performance by administering to a subject an effective amount of the composition of the present invention. Administration can be self-administration.

[0067] The promotion of muscle performance can be one or more of improved muscle function, reduced muscle performance decline, improved muscle strength, improved muscle endurance, and improved muscle recovery.

[0068] Thus, the compositions of the present invention can be used in methods to improve physical endurance (e.g., the ability to perform physical tasks such as exercise, physical labor, sporting activities, etc.), inhibit or delay physical fatigue, promote work capacity, promote endurance, and / or reduce muscle fatigue.

[0069] Improving muscle function can be particularly beneficial for elderly subjects whose muscle function has decreased as a result of aging or age-related conditions.For example, subjects who can benefit from improving muscle function may experience a decrease in muscle function that leads to pre-frailty and frailty.Such subjects may not necessarily experience muscle wasting in addition to the decrease in muscle function.Some subjects, such as subjects with sarcopenia, experience both muscle wasting and decreased muscle function.The compositions of the present invention can be used to promote muscle performance by administering the compositions of the present invention to subjects who are frail or pre-frail.

[0070] In a further embodiment, in the method for promoting muscle performance, the subject suffers from age-related muscle decline, age-related sarcopenia, age-related muscle wasting, physical fatigue, muscle fatigue, and / or is frail or pre-frail. In a further embodiment of the method for promoting muscle performance, the subject suffers from physical fatigue or muscle fatigue, and / or the subject is frail or pre-frail.

[0071] Muscle performance can be sports performance, i.e., the ability of an athlete's muscles to perform when participating in sports activities. Promotion of sports performance, strength, speed, and endurance is measured by increasing muscle contraction strength, increasing muscle contraction amplitude, or shortening muscle reaction time between stimulation and contraction. Athletes refer to individuals who participate in sports at any level and aim to achieve improved strength, speed, or endurance levels in their performance, such as bodybuilders, cyclists, long-distance runners, and sprinters. Promotion of sports performance is manifested by the ability to overcome muscle fatigue, maintain activity for a longer period of time, and perform more effective training.

[0072] The method, medicament or combination of the present invention can improve physical performance in individuals with diseases, including young and elderly people.The composition of the present invention can improve physical performance, for example, short-term performance or long-term performance, in healthy individuals, including athletes, non-athletes, sedentary individuals, and elderly people.This improvement in performance can be measured by the time it takes to walk or run a certain distance (e.g., improvement in performance during the 6-minute walk test (MWT)), improvement in the time it takes to run a certain distance, improvement in IPAQ score on the International Standardized Physical Activity Questionnaire, increase in the number of chair rises within a certain period of time, or another test designed to measure physical performance.

[0073] According to a further embodiment of the present invention there is provided a composition of the present invention for use in a method of promoting physical performance in a subject, for example an elite or sub-elite athlete.

[0074] In certain embodiments, promoting physical performance comprises at least one effect selected from the group consisting of promoting exercise performance, promoting running performance, promoting muscle performance, promoting aerobic endurance, promoting perceived exertion, reducing post-exercise fatigue, promoting muscle recovery, reducing exercise-induced muscle damage, reducing muscle soreness, and promoting exercise-induced muscle damage repair. In further embodiments, promoting physical performance comprises promoting muscle performance during high-intensity aerobic activity. In even further embodiments, promoting physical performance comprises increasing aerobic endurance during high-intensity aerobic activity. In still further embodiments, promoting physical performance comprises increasing resting metabolic rate (RMR). In certain embodiments, promoting physical performance results in improved exercise performance. In further embodiments, promoting physical performance results in improved completion time in a running race. In even further embodiments, promoting physical performance results in a decrease in perceived exertion (RPE).

[0075] In one embodiment, promoting physical performance includes promoting muscle performance during high-intensity aerobic activity.

[0076] In one embodiment, promoting physical performance includes increasing aerobic endurance during high-intensity aerobic activity.

[0077] In one embodiment, promoting physical performance comprises increasing resting metabolic rate (RMR).

[0078] In one embodiment, promoting physical performance results in improved athletic performance.

[0079] In one embodiment, promoting physical performance results in improved completion times for a running race.

[0080] In one embodiment, promoting physical performance results in a decrease in rate of perceived exertion (RPE).

[0081] According to a further embodiment of the present invention there is provided a composition of the present invention for use in a method of promoting physical recovery in a subject, for example an elite or sub-elite athlete.

[0082] In certain embodiments, physical recovery is promoted after high-intensity aerobic activity. In further embodiments, promoting physical recovery comprises at least one effect selected from the group consisting of promoting muscle recovery, promoting exercise performance, promoting running performance, promoting muscle performance, promoting aerobic endurance, promoting perceived exertion, reducing post-exercise fatigue, promoting muscle recovery, reducing exercise-induced muscle damage, reducing muscle soreness, and promoting exercise-induced muscle damage repair. In even further embodiments, promoting physical recovery comprises promoting muscle recovery after high-intensity aerobic activity. In still further embodiments, promoting physical recovery comprises reducing muscle soreness after high-intensity aerobic activity. In certain embodiments, promoting physical recovery comprises reducing the area under the plasma concentration-time curve (AUC) of creatine kinase (CK). CK In a further embodiment, promoting recovery includes reducing CK levels in a subject after aerobic activity compared to baseline, as measured by area under the plasma concentration-time curve (AUC CRP The present invention relates to a method for treating aerobic activity, the method comprising: reducing C-reactive protein (CRP) levels in a subject after aerobic activity compared to baseline, as measured by aerobic activity monitoring.

[0083] In one embodiment, the enhanced physical recovery is after high intensity aerobic activity.

[0084] In one embodiment, promoting physical recovery includes promoting muscle recovery after high-intensity aerobic activity.

[0085] In one embodiment, promoting physical recovery includes reducing muscle soreness following high-intensity aerobic activity.

[0086] In one embodiment, promoting physical recovery is achieved by increasing the area under the plasma concentration-time curve (AUC) of creatine kinase (CK). CK The present invention includes reducing CK levels in a subject after aerobic activity compared to baseline, as measured by CK analysis.

[0087] In one embodiment, promoting recovery is achieved by increasing the area under the plasma concentration-time curve (AUC CRP The present invention includes reducing C-reactive protein (CRP) levels in a subject after aerobic activity compared to baseline as measured by aerobic exercise.

[0088] According to a further embodiment of the present invention there is provided a composition of the present invention for use in a method of promoting endurance in a subject, for example an elite or sub-elite athlete.

[0089] The compositions of the present invention further provide improvements in endurance capacity. Endurance capacity refers to the time to fatigue when exercising at a given workload, typically at an intensity less than 80% of VO2max. The compositions of the present invention can improve endurance capacity in individuals with diseases, including young and elderly individuals. The compositions of the present invention can improve endurance capacity in healthy individuals, including athletes, non-athletes, sedentary individuals, and elderly individuals. The present invention provides a method for increasing the time to fatigue during specific activities, such as fitness training, walking, running, swimming, or cycling. This improvement in endurance capacity can be assessed by objective measures (e.g., speed, oxygen consumption, or heart rate) or by self-report measures (e.g., using a validated questionnaire).

[0090] In certain embodiments, the method includes promoting physical endurance during high-intensity aerobic activity. In further embodiments, promoting physical endurance includes at least one effect selected from the group consisting of promoting muscle recovery, promoting exercise performance, promoting running performance, promoting muscle performance, promoting aerobic endurance, promoting perceived exertion, reducing post-exercise fatigue, promoting muscle recovery, reducing exercise-induced muscle damage, reducing muscle soreness, and promoting repair of exercise-induced muscle damage. In yet further embodiments, promoting physical endurance includes promoting muscle endurance. In still further embodiments, promoting physical endurance includes increasing maximal oxygen consumption (VO2) in a subject. 2max ) is increased.

[0091] In one embodiment, the enhanced physical recovery is during high-intensity aerobic exercise.

[0092] In one embodiment, promoting physical endurance includes promoting muscular endurance.

[0093] In one embodiment, promoting physical endurance comprises increasing maximal oxygen consumption (VO2max) in a subject.

[0094] In certain embodiments, the methods of the present disclosure comprise administering an effective amount of a composition of the present invention to a subject in need thereof, wherein the subject is an elite athlete or a sub-elite athlete. In some such embodiments, the subject is an elite athlete. In certain embodiments, the subject has a VO2 of greater than 65 mL kg min. 2max In further embodiments, the subject has a personal best 3 km running time of less than 9 minutes. In other embodiments, the subject is a sub-elite athlete. In certain such embodiments, the subject has a VO2 of about 60 mL kg min to about 65 mL kg min. 2maxIn a further embodiment, the subject has a personal best 3 km running time of about 9 minutes to about 10 minutes. In certain embodiments, the subject is between about 18 and about 45 years old.

[0095] In some embodiments, the methods, medicaments, or combinations of the present invention find use in reducing inflammation and / or improving resistance to inflammation.

[0096] In some embodiments, the methods, agents, or combinations of the present invention are for the treatment of cognitive impairment. In some embodiments, the methods, agents, or combinations of the present invention improve cardiac efficiency. In some embodiments, the methods, agents, or combinations of the present invention improve metabolic efficiency of the brain. In some embodiments, the methods, agents, or combinations of the present invention reduce the effects of neurodegenerative disorders.

[0097] In some embodiments, the methods, medicaments or combinations of the present invention are for the treatment of Alzheimer's disease or Parkinson's disease.

[0098] In some embodiments, the methods, medicaments or combinations of the present invention are for the treatment of muscle disorders or fatigue.

[0099] In some embodiments, the methods, medicaments or combinations of the present invention maintain or improve cognitive function under fatigue, or reduce the adverse effects on cognitive function under fatigue.

[0100] dosage: The effective amount of the components of the combination to be administered will vary depending on the mode of administration, the age, weight, and health condition of the subject. Factors such as the medical condition, age, and weight of the subject can be important, and the dosage regimen can be adjusted to provide the optimal response.

[0101] The daily dose of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) provided by the compositions of the present invention may be in the range of 20 mg to 5000 mg, for example, 20 mg to 4000 mg, for example, 200 mg to 4000 mg, for example, 20 mg to 3000 mg, for example, 20 mg to 2000 mg, for example, 100 mg to 1000 mg, for example, 100 mg to 800 mg, for example, 200 mg to 600 mg, for example, 200 mg to 400 mg, for example, 200 mg to 300 mg, for example, 250 mg. The daily intake of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) may be provided as a single dose or may be divided among multiple doses.

[0102] Alternatively, the daily dose of ketone bodies (or salts, prodrugs, metabolites or derivatives thereof) provided by the compositions of the present invention may be in the range of 2g to 50g, such as 5g to 30g, for example 10g to 20g, for example 2g, for example 3g, such as 4g, for example 5g, for example 6g, for example 7g, for example 8g, for example 9g, for example 10g, for example 11g, such as 12g, for example 13g, for example 14g, for example 15g, for example 16g, for example 17g, for example 18g, for example 19g, for example 20g, The daily intake of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) may be provided as a single dose or may be divided among multiple doses.

[0103] The unit dose may be in the form of a snack bar, wherein a snack bar weighing 25 g to 150 g, e.g., 40 g to 100 g, may contain the required amount of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) (e.g., 200 mg to 300 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof), or another amount as described above). Alternatively, the unit dose composition may be in the form of a beverage, e.g., provided in a container (e.g., a pouch or bottle) of a volume suitable for a single dose (e.g., 50 mL to 500 mL, e.g., 100 mL to 300 mL). A 100 mL to 300 mL beverage may contain the required amount of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof). Alternatively, the unit dose composition may be in the form of a powder to be reconstituted into a beverage, for example, a quantity of powder suitable for a single dose (e.g., 5 g to 10 g of powder containing 200 mg to 300 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof)). A 100 mL to 500 mL reconstituted beverage may contain the required amount of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof). As described below, compositions for use in the present invention can take any suitable physical form. They may be in the form of a solid (e.g., tablet or bar), semi-solid (e.g., softgel, capsule (e.g., hard capsule), or dragee), powder, or liquid (including emulsion). The composition of the present invention may be a nutritional composition. The composition of the present invention may be a pharmaceutical composition. The composition may be in the form of a dietary supplement, as a functional food, functional beverage, or as a medical food or medical nutritional product.

[0104] The daily intake of a urolithin (or a salt, prodrug, metabolite, or derivative thereof, e.g., urolithin A) component is typically in the range of 10 mg to 5 g per day, for example, 20 mg to 2500 mg per day, for example, 25 mg to 250 mg, for example, 25 mg to 500 mg, for example, 50 mg to 1500 mg per day, for example, 250 mg to 2000 mg, for example, 250 mg to 1500 mg per day, for example, 50 mg to 1000 mg per day, for example, 20 mg to 250 mg per day, for example, 250 mg to 1000 mg per day, for example, 500 mg to 1000 mg per day, for example, 750 mg to 1000 mg per day. In one embodiment, the composition is taken in an amount that provides a dosage of urolithin (or a salt, prodrug, metabolite, or derivative thereof) ranging from about 0.2 mg / kg / day to greater than about 100 mg / kg / day.For example, the dosage of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) may be 0.2 mg / kg / day to 100 mg / kg / day, 0.2 mg / kg / day to 50 mg / kg / day, 0.2 mg / kg / day to 25 mg / kg / day, 0.2 mg / kg / day to 10 mg / kg / day, 0.2 mg / kg / day to 7.5 mg / kg / day, 0.2 mg / kg / day to 5 mg / kg / day, 0.25 mg / kg / day to 100 mg / kg / day, 0.25 mg / kg / day to 25 mg / kg / day, 0.25 mg / kg / day~25mg / kg / day, 0.25mg / kg / day~10mg / kg / day, 0.25mg / kg / day~7.5mg / kg / day, 0.25mg / kg / day~5mg / kg / day, 0.5mg / kg / day~50mg / kg / day, 0.5mg / k g / day~25mg / kg / day, 0.5mg / kg / day~20mg / kg / day, 0.5mg / kg / day~15mg / kg / day, 0.5mg / kg / day~10mg / kg / day, 0.5mg / kg / day~7.5mg / kg / day, 0.5mg / kg / day~ 5mg / kg / day, 0.75mg / kg / day~50mg / kg / day, 0.75mg / kg / day~25mg / kg / day, 0.75mg / kg / day~20mg / kg / day, 0.75mg / kg / day~15mg / kg / day, 0.75mg / kg / day~1 0mg / kg / day, 0.75mg / kg / day ~ 7.5mg / kg / day, 0.75mg / kg / day ~ 5mg / kg / day, 1.0mg / kg / day ~ 50mg / kg / day, 1mg / kg / day ~ 25mg / kg / day, 1mg / kg / day ~ 20mg / kg / day The dose may be 1 mg / kg / day to 15 mg / kg / day, 1 mg / kg / day to 10 mg / kg / day, 1 mg / kg / day to 7.5 mg / kg / day, 1 mg / kg / day to 5 mg / kg / day, 2 mg / kg / day to 50 mg / kg / day, 2 mg / kg / day to 25 mg / kg / day, 2 mg / kg / day to 20 mg / kg / day, 2 mg / kg / day to 15 mg / kg / day, 2 mg / kg / day to 10 mg / kg / day, 2 mg / kg / day to 7.5 mg / kg / day, or 2 mg / kg / day to 5 mg / kg / day.

[0105] The unit dose may be in the form of a snack bar, and a snack bar weighing 25 g to 150 g (e.g., 40 g to 100 g) may contain the required amount of urolithin (or a salt, prodrug, metabolite, or derivative thereof). Alternatively, the unit dose composition may be in the form of a beverage, for example, provided in a container (e.g., a pouch) of a volume suitable for a single dose (e.g., 100 mL to 300 mL). A 50 mL to 500 mL (e.g., 100 mL to 300 mL) beverage may contain the required amount of urolithin (or a salt, prodrug, metabolite, or derivative thereof). Beverages providing compositions of the invention may contain urolithin (or a salt, prodrug, metabolite, or derivative thereof) at a concentration of 0.1 mg per mL to 50 mg per mL, for example, 0.5 mg per mL to 10 mg per mL, for example, 1 mg per mL to 5 mg per mL. Alternatively, the unit dose may be in the form of one or more solids, such as a compressed tablet. A single compressed tablet may contain, for example, a 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1000 mg dose of urolithin (or a salt, prodrug, metabolite, or derivative thereof). Alternatively, the unit dose may be in the form of one or more semi-solid doses, such as a softgel or paste. A single softgel capsule may contain, for example, a dose of 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg, e.g., 250 mg, of a urolithin (or a salt, prodrug, metabolite, or derivative thereof).

[0106] The weight ratio between the ketone body (or salt, prodrug, metabolite, or derivative thereof) and the urolithin (or salt, prodrug, metabolite, or derivative thereof) is generally 500:1 to 1:250, for example, 400:1 to 1:250, for example, 300:1 to 1:250, for example, 300:1 to 1:100, for example, 200:1 to 1:100, for example, 200:1 to 1:75, for example For example, 200:1 to 1:50, for example, 200:1 to 1:25, for example, 50:1 to 1:20, for example, 25:1 to 1:15, for example, 10:1 to 1:10, for example, 5:1 to 1:8, for example, 5:1 to 1:5, for example, 3:1 to 1:5, for example, 1:1 to 1:8, for example, 1:1 to 1:5, for example, 1:2 to 1:6, for example, 1:2 to 1:5, for example, 1:3 to 1:4. The ratio may also be, for example, 1:3 to 5:1, for example, 1:1 to 8:1, for example, 1:1 to 5:1, for example, 2:1 to 6:1, for example, 2:1 to 5:1, for example, 3:1 to 4:1.

[0107] Thus, the compositions of the present invention may comprise 20 mg to 5000 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and 10 mg to 5 g of a urolithin (or salts, prodrugs, metabolites, or derivatives thereof); for example, 20 mg to 4000 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and 10 mg to 5 g of a urolithin (or salts, prodrugs, metabolites, or derivatives thereof); for example, 20 mg to 3000 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and 10 mg to 5 g of a urolithin (or salts, prodrugs, metabolites, or derivatives thereof). or derivatives) and 10 mg to 5 g of a urolithin (or a salt, prodrug, metabolite, or derivative thereof); for example, 20 mg to 2000 mg of a ketone body (or a salt, prodrug, metabolite, or derivative thereof) and 10 mg to 5 g of a urolithin (or a salt, prodrug, metabolite, or derivative thereof); for example, 150 mg to 3000 mg of a ketone body (or a salt, prodrug, metabolite, or derivative thereof) and 10 mg to 5 g of a urolithin (or a salt, prodrug, metabolite, or derivative thereof); for example, 50 mg to 500 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and 50 mg to 500 mg of urolithin (or salts, prodrugs, metabolites, or derivatives thereof); for example, 250 mg to 2000 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and 10 mg to 3000 mg of urolithin (or salts, prodrugs, metabolites, or derivatives thereof); for example, 300 mg to 3000 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and 10 mg to 3000 mg of urolithin (or a salt, prodrug, metabolite, or derivative thereof); for example, 200 mg to 600 mg of ketone bodies (or a salt, prodrug, metabolite, or derivative thereof) and 20 mg to 2500 mg of urolithin (or a salt, prodrug, metabolite, or derivative thereof); for example, 250 mg to 2500 mg of ketone bodies (or a salt, prodrug, metabolite, or derivative thereof) and 100 mg to 2000 mg of urolithin (or a salt, prodrug, metabolite, or derivative thereof);For example, the composition may contain 200 mg to 400 mg of ketone bodies (or their salts, prodrugs, metabolites, or derivatives) and 20 mg to 2500 mg of urolithin (or their salts, prodrugs, metabolites, or derivatives); 200 mg to 300 mg of ketone bodies (or their salts, prodrugs, metabolites, or derivatives) and 50 mg to 1000 mg of urolithin (or their salts, prodrugs, metabolites, or derivatives); 200 mg to 300 mg of ketone bodies (or their salts, prodrugs, metabolites, or derivatives) and 50 mg to 500 mg of urolithin (or their salts, prodrugs, metabolites, or derivatives); or 200 mg to 300 mg of ketone bodies (or their salts, prodrugs, metabolites, or derivatives) and 100 mg to 500 mg of urolithin (or their salts, prodrugs, metabolites, or derivatives). The composition may further contain, for example, proteins, carbohydrates, vitamins, and minerals. ;

[0108] The dosage of compound is referred to herein on a daily dosage basis.In many cases, the beneficial effect of the present composition is most pronounced when the composition is taken for a long period of time, for example, for example, 2 weeks or more, for example, 4 weeks or more, for example, 6 weeks or more, for example, 8 weeks or more, for example, 12 weeks or more, for example, 16 weeks or more, for example, 20 weeks or more, for example, 24 weeks or more.

[0109] In the methods of the invention, it is not necessary for the urolithin (or its salt, prodrug, metabolite, or derivative) and ketone bodies (or its salt, prodrug, metabolite, or derivative) to be administered simultaneously as part of a single composition. The urolithin (or its salt, prodrug, metabolite, or derivative) and ketone bodies (or its salt, prodrug, metabolite, or derivative) can be administered simultaneously or at intervals. The urolithin (or its salt, prodrug, metabolite, or derivative) can be administered first, followed by the ketone bodies (or their salts, prodrugs, metabolites, or derivatives). Alternatively, the ketone bodies (or their salts, prodrugs, metabolites, or derivatives) can be administered first, followed by the urolithin (or its salts, prodrugs, metabolites, or derivatives). The invention further provides kits comprising a urolithin (or its salt, prodrug, metabolite, or derivative) and ketone bodies (or their salts, prodrugs, metabolites, or derivatives), e.g., for use in the methods of the invention. The urolithin (or its salt, prodrug, metabolite, or derivative) and the ketone body (or its salt, prodrug, metabolite, or derivative) may be in different physical forms.

[0110] Preferably, the urolithin (or salt, prodrug, metabolite, or derivative thereof) and ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) are administered simultaneously as part of a single composition. Accordingly, the present invention also provides compositions comprising a urolithin (or salt, prodrug, metabolite, or derivative thereof) and a ketone body (or salt, prodrug, metabolite, or derivative thereof). The compositions of the present invention can be used in place of the combinations of the present invention in all of the embodiments described herein. Furthermore, all disclosures regarding urolithin (or salt, prodrug, metabolite, or derivative thereof) and ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and combinations thereof (e.g., dosages) apply to both the combinations of the present invention and the compositions of the present invention.

[0111] In some embodiments, compositions of the invention comprise a naturally occurring urolithin (or a salt, prodrug, metabolite, or derivative thereof). In some embodiments, compositions of the invention comprise a compound of formula (I) or a salt, prodrug, metabolite, or derivative thereof. In some embodiments, compositions of the invention comprise a compound of formula (I) selected from compounds of formula (I) or a salt, prodrug, metabolite, or derivative thereof, wherein: -Z is OH, W, X, and Y are all H; -A, B, W, X, and Y are all H, and C, D, and Z are all OH (i.e., urolithin C); -A, B, C, W, X, and Y are all H, and D and Z are both OH (i.e., urolithin A).

[0112] In some embodiments, the compositions of the invention comprise urolithin A, urolithin B, urolithin C and / or urolithin D, or a salt, prodrug, metabolite, or derivative thereof.

[0113] In some embodiments, a composition of the invention comprises urolithin A or a salt, prodrug, metabolite, or derivative thereof.

[0114] In some embodiments, the compositions of the invention comprise urolithin A.

[0115] In some embodiments, compositions of the invention comprise two or more urolithins (or salts, prodrugs, metabolites, or derivatives thereof).

[0116] In some embodiments, the compositions of the present invention comprise naturally occurring ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof). In some embodiments, the compositions of the present invention comprise acetoacetic acid, or a salt, prodrug, metabolite, or derivative thereof. In some embodiments, the compositions of the present invention comprise β-hydroxybutyric acid, or a salt, prodrug, metabolite, or derivative thereof. In some embodiments, the compositions of the present invention comprise acetone, or a salt, prodrug, metabolite, or derivative thereof. In some embodiments, the compositions of the present invention comprise acetoacetic acid (or a salt, prodrug, metabolite, or derivative thereof) and β-hydroxybutyric acid (or a salt, prodrug, metabolite, or derivative thereof). In some embodiments, the compositions of the present invention comprise acetoacetic acid (or a salt, prodrug, metabolite, or derivative thereof) and acetone (or a salt, prodrug, metabolite, or derivative thereof). In some embodiments, the compositions of the present invention comprise β-hydroxybutyric acid (or a salt, prodrug, metabolite, or derivative thereof) and acetone (or a salt, prodrug, metabolite, or derivative thereof). In some embodiments, the compositions of the present invention comprise acetoacetic acid (or a salt, prodrug, metabolite, or derivative thereof), beta-hydroxybutyric acid (or a salt, prodrug, metabolite, or derivative thereof), and acetone (or a salt, prodrug, metabolite, or derivative thereof). In some embodiments, the compositions of the present invention comprise acetoacetic acid. In some embodiments, the compositions of the present invention comprise beta-hydroxybutyric acid. In some embodiments, the compositions of the present invention comprise acetone. In some embodiments, the compositions of the present invention comprise acetoacetic acid and beta-hydroxybutyric acid. In some embodiments, the compositions of the present invention comprise acetoacetic acid and acetone. In some embodiments, the compositions of the present invention comprise beta-hydroxybutyric acid and acetone. In some embodiments, the compositions of the present invention comprise acetoacetic acid, beta-hydroxybutyric acid, and acetone.

[0117] In some embodiments, the compositions of the present invention comprise two or more ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof).

[0118] Form of composition: The compositions of the present invention can take any suitable physical form. They may be in the form of a solid (e.g., a tablet or bar), a semi-solid (e.g., a softgel, a capsule (e.g., a hard capsule), or a dragee), a powder, or a liquid (including an emulsion). The compositions of the present invention may be nutritional compositions. The compositions of the present invention may be pharmaceutical compositions. The compositions may be in the form of a dietary supplement, as a functional food, functional drink, or as a medical food or medical nutritional product.

[0119] The tablet-form compositions may be of any suitable type and may contain conventional excipients in the art. The excipients may, for example, provide the desired hardness, shelf life, and flavor so that the composition has an acceptable taste, an attractive appearance, and good storage stability. The bar may be of any suitable type and may contain ingredients conventionally used for preparing snack bars.

[0120] The snack bar may be a unit dose of the composition of the invention, and a snack bar weighing 25 g to 150 g, for example, 40 g to 100 g, may contain the required amount of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) as described above, and the required amount of urolithin (or salts, prodrugs, metabolites, or derivatives thereof) as described above.

[0121] The semi-solid form may also contain conventional excipients in the art. The excipients can, for example, provide the desired hardness, shelf life, and flavor so that the composition has an acceptable taste, an attractive appearance, and good storage stability. The semi-solid form may be provided for oral or topical administration.

[0122] Powders are commonly used to deliver nutritional and medical compositions. Powders have the advantage that multiple doses can be provided in a single container, allowing various dose sizes to be used from the same delivery container. Powders generally have good storage properties. Powder compositions may also contain excipients conventional in the art. Excipients can provide, for example, shelf life, flavor, and moisture resistance so that the composition has acceptable taste, attractive appearance, and good storage stability. The present invention can take the form of a kit comprising a composition containing ketone bodies (or a salt, prodrug, metabolite, or derivative thereof) together with a separate composition containing a urolithin (or a salt, prodrug, metabolite, or derivative thereof), e.g., a ketone body powder composition together with a separate solid or liquid composition containing a urolithin (or a salt, prodrug, metabolite, or derivative thereof). A solid or liquid composition containing a urolithin (or a salt, prodrug, metabolite, or derivative thereof), such as a tablet or beverage, or other form described herein, may be provided with instructions for use with the ketone powder. For example, both the ketone body (or a salt, prodrug, metabolite, or derivative thereof) and the urolithin (or a salt, prodrug, metabolite, or derivative thereof) may be in powder form.

[0123] The liquid composition may be in the form of a medicine or a drink. The liquid preparation may be a solution, emulsion, slurry, or other semi-liquid. The excipients in the liquid composition may provide, for example, shelf life, appearance, flavor, and texture so that the composition has acceptable taste, attractive appearance, and good storage stability. The liquid composition may be provided for oral administration. The liquid composition may be provided for topical application in the form of, for example, a cream, ointment, or lotion.

[0124] The beverage may be a unit dose of the composition of the present invention and may be provided, for example, in a container (e.g., a pouch or bottle) of a volume suitable for a single dose (e.g., 50 mL to 500 mL, e.g., 100 mL to 300 mL). A 100 mL to 300 mL beverage may contain the required amount of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) as described above, and the required amount of urolithin (or salts, prodrugs, metabolites, or derivatives thereof) as described above.

[0125] Alternatively, the powder to be reconstituted into a beverage may be a unit dose of the composition of the invention, for example, a quantity of powder suitable for a single dose (e.g., 5 g to 10 g of powder containing 200 mg to 300 mg of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof)). A 100 mL to 500 mL reconstituted beverage may contain the required amount of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) as described above, and the required amount of urolithin (or salts, prodrugs, metabolites, or derivatives thereof) as described above.

[0126] For some applications, the compositions of the present invention may also be in the form of a solution suitable for injection or intravenous administration.

[0127] Additional ingredients in the compositions of the present invention: Compositions according to the present invention may contain additional ingredients in addition to a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof). The additional ingredients may be compounds that provide health benefits, for example, selected from medium-chain triglycerides, phospholipids (e.g., phosphatidylcholine), vitamins, minerals, polyunsaturated fatty acids, functional amino acids, and other compounds.

[0128] The European Pharmacopoeia describes medium-chain triglycerides as solidified oils extracted from the hard, dry parts of the endosperm of Cocos nucifera L. (coconut palm) or from the dried endosperm of Elaeis guineenis Jacq. (African oil palm). Both the European Pharmacopoeia and USPNF have specifications for medium-chain triglycerides that require the presence of certain fatty acids as follows: caproic acid (C6) ≤ 2.0%, caprylic acid (C8) 50.0-80.0%, capric acid (C10) 20.0-50.0%, lauric acid (C12) ≤ 3.0%, and myristic acid (C14) ≤ 1%.

[0129] Medium chain triglycerides for use in preferred compositions include a mixture of fatty acid chains and triglycerides present in the following ratios: C6≦5%, C8 50-70%, C10 30-50%, and C12≦12%, for example, C6≦0.5%, C8 55-65%, C10 35-45%, and C12≦1.5%.

[0130] The medium chain triglycerides used in the preferred compositions may be derived from any known or otherwise suitable source.

[0131] Among the vitamins, specific mention may be made of vitamin A, vitamin C, vitamin D, vitamin E, vitamin B12, and vitamin K2. As used herein, "vitamin D" refers to any of the known forms of vitamin D, specifically including vitamin D2 (ergocalciferol), vitamin D3 (cholecalciferol), vitamin D precursors, metabolites, and other analogs, and combinations thereof, as well as various active and inactive forms of vitamin D. For example, vitamin D3 may be provided in its non-hydroxylated, inactive form as cholecalciferol, or in its hydroxylated, active form as calcitriol.

[0132] Creatine has been described as having beneficial effects in the treatment of muscle disorders and can be included in the compositions of the present invention. β-Hydroxyl-β-methylbutyrate (HMB) has been described as having beneficial effects in the treatment of muscle disorders and can be included in the compositions of the present invention.

[0133] Among the minerals, particular mention may be made of calcium salts (for example calcium phosphate), selenium, zinc salts, magnesium salts, and iron salts.

[0134] Polyunsaturated fatty acids are fatty acids that contain two or more double bonds in the backbone. This class includes many important compounds, such as essential fatty acids, e.g., omega-3 and omega-6 fatty acids. Long-chain polyunsaturated fatty acids, preferably those with at least 20 carbon atoms in the molecule, are preferred. Such long-chain omega-3 fatty acids include cis-11,14,17-eicosatrienoic acid (ETE) C20:3, cis-8,11,14,17-eicosatetraenoic acid (ETA) C20:4, cis-5,8,II,14,17-eicosapentaenoic acid (EPA) C20:5, cis-7,10,13,16,19-docosapentaenoic acid (DPA, clupanodonic acid) C22:5, cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) C22:6, cis-9,12,15,18,21-tetracosapentaenoic acid C24:5, and cis-6,9,12,15,18,21-tetracosahexaenoic acid (Nisinic acid) C24:6. Long-chain omega-6 fatty acids having at least 20 carbon atoms include cis-11,14-eicosadienoic acid C20:2, cis-8,11,14-eicosatrienoic acid (dihomo-γ-linolenic acid) (DGLA) C20:3, cis-5,8,11,14-eicosatetraenoic acid (arachidonic acid) (AA) C20:4, cis-13,16-docosadienoic acid C22:2, cis-7,10,13,16-docosatetraenoic acid (adrenic acid) C22:4, and cis-4,7,10,13,16-docosapentaenoic acid (osbondoic acid) C22:5. The compositions according to the present invention preferably contain EPA, DHA, or a combination thereof, for example, in an amount of 10 mg to 1000 mg per serving, e.g., 25 mg to 250 mg per serving. For many muscle growth and / or muscle-promoting treatments, it is beneficial to provide certain specific amino acids. For example, L-arginine, L-glutamine, lysine, and branched-chain amino acids are considered important. These amino acids are sometimes known as "functional amino acids." The compositions of the present invention may include one or more branched-chain amino acids (leucine, isoleucine, and valine). The compositions of the present invention may include one or both of L-arginine and L-glutamine. The compositions of the present invention may also include lysine.

[0135] The pharmaceutical compositions of the invention may include an additional pharmaceutically active compound, such as a statin. The invention may be provided as a kit comprising a composition of a urolithin (or a salt, prodrug, metabolite, or derivative thereof), a ketone body (or a salt, prodrug, metabolite, or derivative thereof), and a pharmaceutically active compound, such as a statin.

[0136] The compositions of the present invention may also contain one or more additional agents useful in the treatment of mitochondrial biogenesis or mitochondrial disorders, including, but not limited to, resveratrol, pyrroloquinoline quinone, ubiquinone, sulforaphane, coenzyme Q10, genistein, hydroxyl tyrosol, quercetin, L-carnitine, alpha-lipoic acid, and folinic acid (e.g., leucovorin).

[0137] Additional compounds may also (or alternatively) be included in the compositions of the present invention, including, for example, tomatidine, ursolic acid, curcumin, capsaicin, menthol, trolamine salicylate, and methyl salicylate.

[0138] The additional compound may be an additional active ingredient or supplement, such as carnitine or its salt. The term carnitine includes L-carnitine and its derivatives, including acetyl-L-carnitine (ALCAR) and propionyl-L-carnitine. Carnitine salts include tartrates, such as L-carnitine L-tartrate (LCLT), and glycine salts, such as glycine propionyl-L-carnitine (GPLC). When used, carnitine may be administered by any suitable means or dosage form, but typically carnitine is administered orally and daily.

[0139] In some exemplary embodiments, compositions of the present disclosure may include one or more ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) and urolithins (or salts, prodrugs, metabolites, or derivatives thereof), as well as one or more additional macronutrients, typically protein, fat, or carbohydrate, or two or more of protein, fat, and carbohydrate.

[0140] Any suitable fat or oil source of the type commonly used in food and pharmaceutical preparations can be used in the compositions of the present invention.Non-limiting examples of fat sources suitable for use in the compositions described herein include coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm oil and palm kernel oil, palm olein, canola oil, fish oil, cottonseed oil, polyunsaturated fatty acids such as docosahexaenoic acid (DHA), arachidonic acid (ARA), eicosapentaenoic acid (EPA), and combinations thereof.

[0141] Non-limiting examples of carbohydrates or their sources suitable for use in the compositions described herein may include maltodextrin, hydrolyzed or modified starch or corn starch, glucose polymers, corn syrup, corn syrup solids, carbohydrates derived from rice, glucose, fructose, lactose, trehalose, high fructose corn syrup, tapioca dextrin, isomaltulose, sucromalt, maltitol powder, glycerin, fructooligosaccharides, soy fiber, corn fiber, guar gum, konjac flour, polydextrose, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), and combinations thereof.Maltodextrin, sucrose, and fructose are particularly preferred.

[0142] Non-limiting examples of proteins or sources thereof suitable for use in the compositions described herein may include hydrolyzed, partially hydrolyzed, or non-hydrolyzed proteins or protein sources. They may be derived from any known or otherwise suitable source, such as milk (e.g., casein, whey), animal (e.g., meat, fish), grain (e.g., rice, corn), or plant (e.g., soy, pea) sources. Combinations of protein sources or types may also be used. Non-limiting examples of proteins or sources thereof include intact pea protein, intact pea protein isolate, intact pea protein concentrate, milk protein isolate, milk protein concentrate, casein protein isolate, casein protein concentrate, whey protein isolate, sodium or calcium casemate, whole milk, partially or fully skimmed milk, yogurt, soy protein isolate, and soy protein concentrate, as well as combinations thereof. Combinations of protein sources or types may also be used. For example, Greek and Icelandic yogurts are generally known to have particularly high protein contents, making them particularly suitable for use in the formulations of the present invention. Yogurts for use in the compositions of the present invention may contain, for example, 2 g to 15 g of protein per 100 g. Particularly preferred are yogurts with a high protein content, for example, 6 g to 15 g per 100 g, e.g., 7 g to 15 g per 100 g, e.g., 8 g to 15 g per 100 g. Optionally, supplemental proteins may also be added to the yogurt formulation to increase the protein content of the formulation. Yogurts of the present invention may also contain live cultures, such as S. thermophilus, L. bulgaricus, L. acidophilus, or L. lactis.

[0143] The total concentration or amount of protein, fat, carbohydrates, and other ingredients will vary depending on the nutritional needs of the intended user.

[0144] The additional components in the compositions of the present invention may be compounds that do not provide a health benefit to the subject, but instead improve the composition in some other way, for example, its taste, texture, or shelf life, as described above. Thus, the compositions of the present invention may further contain one or more compounds selected from emulsifiers, colorants, preservatives, gums, hardening agents, thickeners, sweeteners, and flavorings.

[0145] Suitable emulsifiers, colorants, preservatives, gums, hardeners, and thickeners are well known in the art of emulsion and other semi-liquid manufacturing. For example, preservatives such as benzoic acid, sorbic acid, phosphoric acid, lactic acid, acetic acid, hydrochloric acid, and their soluble salts may be used.

[0146] The emulsifier may include one or more of phosphatidylcholine, lecithin, polysorbate such as polysorbate 60 or polysorbate 80 (Tween-60 and Tween-80), and glycerol monostearate (GMS), which is also known as glyceryl monostearate.

[0147] Stabilizers may be used in the compositions described herein. Many compositions are stable suspensions without the need for additional stabilizers. A stable suspension is one that does not undergo phase separation over time. For certain compositions, the inclusion of additional stabilizers may improve stability. Stabilizers suitable for use in the compositions of the present invention include glycerol monostearate (GMS), silicon dioxide, and vegetable shortening. An exemplary stabilizer is GMS, and preferred compositions of the present invention contain GMS. Due to its properties, GMS also serves as an excellent solvent for phospholipids, such as those found in lecithin. GMS exists in two polymorphic forms: the α-form, which is dispersible and foamy and is useful as an emulsifier or preservative; the β-form, which is suitable for wax matrices; the α-form is converted to the β-form when heated at 50°C.

[0148] GMS is classified into two distinct grades: 40-55 percent monoglyceride and 90 percent monoglyceride. The 40-55 percent monoglyceride, as defined by the European Pharmacopoeia, describes GMS as a mixture of monoacylglycerols and primarily monostearoylglycerol, along with some di- and triglycerols. Specifically, the 40-55 grade contains 40-55% monoacylglycerol, 30-45% diacylglycerol, and 5-15% triacylglycerol. The 99 percent grade contains over 90% monoglycerides. The monoglycerides in commercially available GMS products are mixtures of various ratios of glyceryl monostearate and glyceryl monopalmitate. The European Pharmacopoeia further divides glyceryl monostearate 40-55 into three types depending on the proportion of stearic acid esters in the mixture. Type 1 contains 40.0-60.0% stearic acid, and the sum of palmitic acid and stearic acid is ≦90%. Type 2 contains 60.0-80.0% stearic acid, and the sum of palmitic acid and stearic acid is ≦90%. Type 3 contains 90.0-99.0% stearic acid, and the sum of palmitic acid and stearic acid is ≦96%. Any form of GMS can be used in the composition.

[0149] In some embodiments, compositions of the invention comprise a urolithin (e.g., urolithin A) or a salt, prodrug, metabolite, or derivative thereof, one or more ketone bodies or a salt, prodrug, metabolite, or derivative thereof, a medium-chain triglyceride, and a stabilizer, such as glycerol monostearate. In some embodiments, compositions of the invention comprise a urolithin (e.g., urolithin A) or a salt, prodrug, metabolite, or derivative thereof, one or more ketone bodies or a salt, prodrug, metabolite, or derivative thereof, a medium-chain triglyceride, an emulsifier, and a stabilizer.

[0150] Metal chelating or sequestrant agents, such as the sodium calcium salt of ethylenediaminetetraacetic acid (EDTA), may also be used in the compositions of the present invention. Other components that may be included in the formulations of the present invention may include polyethylene glycol, silicon dioxide, vegetable shortening, and beeswax.

[0151] Sweeteners may be particularly useful in the compositions of the present invention. For example, strong non-nutritive carbohydrate sweeteners may be used, such as aspartame, sucrose, acesulfame potassium, saccharin, cyclamate, stevia, thaumatin, and mixtures thereof. Aspartame is particularly suitable.

[0152] Flavorings can be particularly useful in the compositions of the present invention.In liquid or semi-liquid compositions, fruit flavors can be provided by including fruit sauce or puree.Typical flavorings include strawberry, raspberry, blueberry, apricot, pomegranate, peach, pineapple, lemon, orange, and apple.Generally, fruit flavorings include fruit extracts, fruit jams, or fruit purees, together with sweeteners, starches, stabilizers, natural and / or artificial flavors, colorants, preservatives, water, and citric acid or other suitable acid combinations for pH control.

[0153] For oral preparations, the compositions can be used alone or in combination with suitable additives to prepare tablets, powders, granules, or capsules, for example, in combination with conventional additives such as lactose, mannitol, corn starch, or potato starch; binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch, or gelatin; disintegrants such as corn starch, potato starch, or sodium carboxymethylcellulose; lubricants such as talc or magnesium stearate; and, if necessary, diluents, buffers, humectants, preservatives, and flavoring agents.

[0154] The compositions can be formulated into liquid preparations by dissolving, suspending, or emulsifying them in aqueous or non-aqueous solvents such as vegetable oils or other similar oils, synthetic fatty acid glycerides, esters of higher fatty acids, or propylene glycol, and can be formulated into liquid preparations using conventional additives such as solubilizers, isotonicity agents, suspending agents, emulsifiers, stabilizers, and preservatives, as needed.The compositions can be used in aerosol preparations administered via inhalation.They can be made into suppositories by mixing with various bases, such as emulsifying bases or water-soluble bases.

[0155] Unit dosage forms for oral administration, such as syrups, elixirs, and suspensions, may be provided, with each dosage unit, e.g., a teaspoon, tablespoon, tablet, or capsule, containing a predetermined amount of the composition of the present invention. Similarly, unit dosage forms for injection or intravenous administration may comprise a compound of the present invention in a composition as a solution in sterile water, saline, or another pharmaceutically acceptable carrier, with each dosage unit, e.g., mL or L, containing a predetermined amount of the composition of the present invention. [Table 2] [Table 3]

[0156] Bulk powders are usually provided with instructions informing the subject how much powder to use per serving. For example, bulk powders may be provided in containers with measuring spoons of the required size that allow the correct amount of powder to be measured out. The powders may be taken as is, mixed with food, or added to water or juice to make a beverage. [Table 4] [Table 5] [Table 6] [Table 7] [Table 8]

[0157] Representative Softgel Capsule Formulation (i.e., a softgel capsule containing a gelatin shell and a fill containing urolithin A) [Table 9] [Table 10]

[0158] The composition of the present invention can be administered as a single treatment, or more generally as a series of treatments.In one example, the subject is administered a dose before or after exercise.For subjects who cannot exercise, the dose of the composition can be administered, for example, once, twice, or three times per day, or once, twice, three, four, five, or six times per week.It will also be understood that the effective dosage of the compound can increase or decrease during the course of a particular treatment. [Example]

[0159] The following examples illustrate the invention.

[0160] Example 1: Effect of the combination of the ketone bodies 3-β-hydroxybutyrate and acetoacetate, and urolithin A, on cellular energy production 3-β-hydroxybutyrate and acetoacetate are ketone bodies that are produced by the liver during ketogenesis and can be used by peripheral tissues to generate ATP. To do this, 3-β-hydroxybutyrate and acetoacetate are imported into the mitochondria and converted to acetyl-CoA. Acetyl-CoA then enters the tricarboxylic acid cycle and can be converted to citrate by citrate synthase. NAD + and reduction of the FADH cofactor, allowing the production of ATP by oxidative phosphorylation.

[0161] C2C12 cells are incubated with 20 μM urolithin A or vehicle, with or without either 3-β-hydroxybutyrate, acetoacetate, or a combination of the two, for 24 hours to assess the effects of 3-β-hydroxybutyrate and acetoacetate, alone or in combination with UA, on mitochondrial function.

[0162] ATP and citrate synthase Total ATP content and citrate synthase (CS) enzyme activity are measured as described in Mouchiroud, L. et al., Cell 154, 430-441 (2013). Briefly, total ATP content is measured by the CellTiter-Glo® Luminescent Cell Viability Assay (Promega), and CS is determined using a CS assay kit (Sigma).

[0163] breathing ability Cells are incubated with test compounds in high glucose medium (normal cell culture conditions) and low glucose conditions using continuous infusion of oligomycin, FCCP (carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone), and rotenone / antimycin, allowing determination of maximal respiratory capacity and uncoupled respiration.

[0164] To distinguish between anaerobic glycolytic ATP and oxidative phosphorylated ATP, the protocol Seahorse XF ATP is also used. (https: / / www.agilent.com / en / products / cell-analysis / seahorse-xf-atp).

[0165] fatty acid oxidation Measurements are performed according to a standard protocol for fatty acid oxidation (https: / / www.agilent.com / cs / pubimages / misc / XF_FAO_Assay_Technical_Brief.pdf). Briefly, once fully differentiated in 96-well plates, C2C12 myotubes are treated for 24 hours, 45 minutes prior to the assay, in a DMEM-based, substrate-defined medium containing 0.5 mM glucose, 1 mM GlutaMAX™, and 0.5 mM carnitine. Cells are washed twice on the day of the assay with assay medium (111 mM NaCl, 4.7 mM KCl, 1.25 mM CaCl, 2 mM MgSO, 1.2 mM NaHPO, 2.5 mM glucose, 0.5 mM carnitine, and 5 mM HEPES, adjusted to pH 7.4 at 37°C). The cells are then incubated for 30 minutes in a non-CO2 incubator. Immediately before starting the assay, the cells are supplemented with XF palmitate-BSA FAO substrate (Seahorse Bioscience) at a final concentration of 166 μM palmitate. Oxygen consumption rates are determined at basal levels and after the addition of FCCP.

[0166] Example 2: Effects of the combination of the ketone bodies 3-β-hydroxybutyrate and acetoacetate, and urolithin A, on gene expression of markers of mitochondrial function C2C12 cells are incubated with 20 μM urolithin A or vehicle (with or without 3-β-hydroxybutyrate, acetoacetate, or a combination of the two) for 24 hours to assess the effects of 3-β-hydroxybutyrate and acetoacetate, alone or in combination with urolithin A, on the expression of genes encoding mitochondrial proteins.

[0167] Gene expression measured by RNA-seq Total RNA from C2C12 cells was extracted using TRIzol (Thermo Fisher Scientific) and purified by column chromatography using the RNeasy® Mini Kit (QIAGEN, 74104). The Agilent 2100 Bioanalyzer (Agilent RNA 6000 Nano Kit) was used to detect total RNA sample concentration, RIN, 28S / 18S, and size. Sample purity was tested using NanoDrop™. RNA-seq library preparation was performed as described in D'Amico et al., Mol Cell 2019. RNA-seq runs were performed using an Illumina HiSeq 4000 sequencing platform with single reads (1 × 50 bp) and 30 million reads (±3%). The resulting reads were first trimmed to remove adapter sequences and then mapped to the human genome (GRCh37). Perform gene set enrichment analysis using the GSEA tool by calculating differential gene expression (DGE) and selecting genes with a nominal p-value less than 0.05. Gene sets are retrieved from the MSIGDB C5 GO collection. Gene sets related to mitochondrial biology with a false discovery rate (FDR) of <0.25 were selected.

[0168] Example 3: Synergistic Anti-inflammatory Effects of Urolithin A and Sodium β DL-β-Hydroxybutyrate (BHB) method C2C12 myoblasts purchased from the American Type Culture Collection (ATCC) (reference number CRL-1772) were seeded at 8,000 cells per well in 96-well plates (Greiner, Kremsmunster, Austria) using DMEM Glutamax 4.5g D-glucose (Gibco, reference number 31966021) supplemented with 10% fetal bovine serum (FBS) (PanBiotech, reference number P190902), 1% PenStrep (Biowest), and 1% HEPES (Biowest). Cells were then differentiated for 6 days using differentiation medium (DMEM Glutamax 4.5g D-glucose (Gibco, reference number 31966021), 2% horse serum (Thermofisher, reference number 160501229), 1% PenStrep, and 1% HEPES). All media were sterilized using a 0.45 μm filter unit (VWR). After 4 days of differentiation, cells were treated with 1) urolithin A (UA), DL-β-hydroxybutyric acid sodium salt (BHB) (Sigma reference number: H6501-5GA) resuspended in DMSO, or a combination of the two at the indicated doses for 24 hours. A cytokine mixture ("cytomix") was added on day 5 for an additional 24 hours in addition to the previous treatment. Each cytokine was used at 5 ng (mIFN-γ (Roche reference number 11276905001), TNF-α (Peprotech reference number 300-01A), and IL-β (Proteintech reference number 200-01B)). The media was collected, and an interleukin-6 ELISA was performed according to the supplier's instructions (Proteintech reference number KE10007). Finally, the absorbance was measured at a wavelength of 450 nm using a microplate plate reader (Fluostar Optima, BMGLabtech).

[0169] result Exposure of C2C12 cells to a mixture of pro-inflammatory cytokines (cytomix) resulted in a significant increase in interleukin-6 (IL-6) secretion, measured as the concentration of IL-6 in the cell culture medium. Urolithin A pretreatment dose-dependently reduced IL-6 secretion, with a 25% dose-dependent reduction at the low dose of 6.25 μM and a 60% reduction at the high dose of 12.5 μM (Figure 1).

[0170] To explore the potential synergistic effects of urolithin A (UA) and β-hydroxybutyrate (BHB), cells were treated with both compounds alone or in combination using a fixed sub-functional UA dose of 6.25 μM and either 5 μM or 25 μM BHB.

[0171] Low doses of BHB (5 μM) alone have no effect on IL-6 secretion (-0.007%). Surprisingly, the combination of low doses of UA (6.25 μM) and BHB (5 μM) results in a significant 56% reduction in inflammatory markers. This is also statistically significant compared to the 25% reduction achieved with UA 6.25 μM alone (Table 1).

[0172] High doses of BHB (2 μM) significantly reduced IL-6 secretion by 32%. The combination of low doses of UA (6.25 μM) and high doses of BHB reduced IL-6 secretion by 58%. This demonstrates the ability of UA at a fixed dose to act synergistically with BHB across a range of doses (Figure 2 and Table 1).

[0173] Ketone bodies such as beta-hydroxybutyrate have been shown to reduce inflammation in certain cell-based models. However, this biological effect of BHB was only observed at high concentrations above 2 mM (PMID:25686106) or 250 μM (PMC6402511). The results of combining 6.25 μM urolithin A with 5 μM BHB demonstrate a specific and surprising dose window in which UA can unlock the potential of otherwise ineffective low BHB doses. [Table 11]

Claims

1. A combination comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

2. A combination consisting of a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

3. A composition comprising a urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

4. The urolithin is a compound of formula (I): 【Chemistry 1】 4. The combination of claim 1 or 2, or the composition of claim 3, wherein A, B, C, D, W, X, Y, and Z are each independently selected from H and OH.

5. 5. The combination or composition of claim 4, wherein the urolithin is selected from urolithin A, urolithin B, urolithin C, and / or urolithin D.

6. 5. The combination or composition of claim 4, wherein the urolithin is urolithin A.

7. 7. The combination or composition of any one of claims 1 to 6, wherein the ketone body is a naturally occurring ketone body (or a salt, prodrug, metabolite, or derivative thereof).

8. 8. The combination or composition of any one of claims 1 to 7, wherein the ketone body is acetoacetic acid, or a salt, prodrug, metabolite, or derivative thereof, and / or β-hydroxybutyric acid, or a salt, prodrug, metabolite, or derivative thereof, and / or acetone, or a salt, prodrug, metabolite, or derivative thereof.

9. The combination or composition according to any one of claims 1 to 8, wherein the ketone bodies are acetoacetic acid and / or β-hydroxybutyric acid and / or acetone.

10. 10. The combination or composition of any one of claims 1 to 9, wherein the ketone body is β-hydroxybutyric acid (or a salt, prodrug, metabolite, or derivative thereof).

11. The combination or composition according to any one of claims 1 to 10, wherein the ketone body is β-hydroxybutyric acid.

12. 12. A method for providing a beneficial health effect to a subject, said method comprising administering to the subject a combination or composition according to any one of claims 1 to 11.

13. 12. A combination or composition according to any one of claims 1 to 11 for use in providing a beneficial health effect to a subject.

14. 12. Use of a combination or composition according to any one of claims 1 to 11 to provide a beneficial health effect to a subject.

15. 12. Use of a combination or composition according to any one of claims 1 to 11 in the manufacture of a medicament for providing a beneficial health effect to a subject.

16. A combination or composition according to any one of claims 1 to 11 for use as a medicament.

17. 12. A combination or composition according to any one of claims 1 to 11 for use in improving muscle performance, maintaining or improving muscle function, preventing the decline of muscle function, increasing muscle mass and / or reducing muscle wasting.

18. 12. A combination or composition according to any one of claims 1 to 11 for use in improving physical endurance, inhibiting or delaying physical fatigue, promoting work capacity, promoting endurance and / or reducing muscle fatigue.

19. A combination or composition according to any one of claims 1 to 11 for use in promoting physical performance.

20. A combination or composition according to any one of claims 1 to 11 for use in reducing inflammation and / or improving resistance to inflammation.

21. A kit comprising: (i) a composition comprising a ketone body, or a salt, prodrug, metabolite, or derivative thereof; (ii) a separate composition comprising a urolithin, or a salt, prodrug, metabolite, or derivative thereof.

22. 22. The kit of claim 21, wherein the urolithin, or salt, prodrug, metabolite, or derivative thereof, is as defined in any one of claims 4 to 6, and / or the ketone body, or salt, prodrug, metabolite, or derivative thereof, is as defined in any one of claims 7 to 11.

23. 23. The kit of claim 21 or 22, wherein the urolithin is urolithin A.

24. The kit according to any one of claims 21 to 23, wherein the ketone body is selected from acetoacetic acid and / or β-hydroxybutyric acid and / or acetone.

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