Combination of urolithin and ketone bodies

Abstract

The present invention relates to methods that involve administering a combination of urolithin and one or more ketones to provide beneficial health effects, such as improved resistance to inflammation, mitochondrial function, and cellular metabolism. The methods may, for example, be used to improve the health and wellness of subjects, particularly elderly or weak persons; and for improving physical fitness, muscle performance and / or endurance of a person engaged in exercise. The methods may also be used to treat or prevent various conditions, such as conditions associated with insufficient mitochondrial activity, excessive inflammation, and / or muscle-related conditions. The invention further relates to compositions comprising urolithin and one or more ketone bodies.

Description

TECHNICAL FIELD

[0001] The present invention relates to methods involving administration of a combination of urolithin and one or more ketone bodies, thereby providing a health benefit, such as improved resistance to inflammation, mitochondrial function and cellular metabolism. The methods can for example be used to improve the health and well-being of a subject, in particular an elderly person or a frail person; and to improve the physical fitness, muscle performance and / or endurance of a person engaged in exercise. The methods can also be used to treat or prevent various conditions, such as conditions associated with insufficient mitochondrial activity, excessive inflammation and / or muscle-related disorders. The invention further relates to compositions comprising a combination of urolithin and one or more ketone bodies. BACKGROUND

[0002] Urolithins have been proposed for use in the treatment of various conditions associated with insufficient mitochondrial activity, including obesity, decreased metabolic rate, metabolic syndrome, diabetes, cardiovascular disease, hyperlipidemia, neurodegenerative disease, cognitive impairment, mood disorders, stress and anxiety; for weight management or for improving muscle performance or mental performance. See WO2012 / 088519 (Amazentis SA). In WO2007 / 127263 (The Regents of the University of California), the use of urolithins for the treatment of various neoplastic diseases is described.

[0003] International Patent Publication WO 2014 / 004902 (originating from application PCT / US2013 / 48310) discloses a method of increasing autophagy, including mitochondrial autophagy, in a cell, the method comprising contacting the cell with an effective amount of a urolithin or a pharmaceutically acceptable salt thereof, thereby increasing autophagy, including mitochondrial autophagy, in the cell. The subject can be administered who has a disease or condition selected from the group consisting of metabolic stress, cardiovascular disease, endothelial cell dysfunction, muscle wasting, 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, growth failure, loss of muscle coordination, muscle weakness, vision problems, hearing problems, heart disease, liver disease, kidney disease, gastrointestinal disorders, respiratory distress, neurological problems, autonomic dysfunction (sometimes learning disabilities), and dementia (caused by mitochondrial disease), muscle disease, cancer, cognitive impairment, stress, and mood disorders.

[0004] In particular, urolithins have been proposed to treat muscle-related pathological conditions. Muscle-related pathological conditions include myopathies and neuromuscular diseases. Examples of such conditions include Duchenne muscular dystrophy, acute muscle wasting, e.g., muscle atrophy and / or cachexia associated with, e.g., burns, bed rest, limb immobilization, or major thoracic, abdominal, neck, and / or orthopedic surgery, amyotrophic lateral sclerosis, and multiple sclerosis. Age-related muscle loss is a particularly prevalent condition. Cachexia due to long-term immobility or other diseases, such as cancer, is another condition often characterized by poor muscle performance.

[0005] Another biological function of urolithin A is to attenuate harmful inflammatory responses. This is particularly important in the clinic because aging and most age-related diseases are associated with chronic, low-grade inflammation. This process has recently been named inflammatory aging, which contributes to age-related decline in cellular and organismal function.

[0006] Effective muscle function and physical performance is important for healthy individuals as well as individuals with disease, 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, shortening the muscle reaction time between stimulation and contraction, are all beneficial to individuals, especially athletes. For the elderly with age-related decline in muscle function, including muscle loss / wasting, or individuals with cachexia muscle wasting, improvement in muscle and physical performance is important for basic aspects of daily functioning, such as walking speed and the distance they walk without assistance.

[0007] Ketones are a class of carbonyl compounds, including acetoacetate, beta-hydroxybutyrate (BHB) and acetone, which are produced by the liver from fatty acids under certain conditions. The ketones are known to induce mitochondrial biogenesis and are believed to be beneficial for muscle performance, metabolism, neuroprotection, healthy aging, cardiovascular health, cognitive performance and mental health. Ketones are also believed to be useful in the treatment of cancer. SUMMARY

[0008] The present invention provides a combination comprising or consisting of urolithin (or a salt, prodrug, metabolite or derivative thereof) and a ketone (or a salt, prodrug, metabolite or derivative thereof).

[0009] The present invention provides a method for providing a beneficial health effect in a subject, the method comprising administering to the subject a combination comprising or consisting of urolithin (or a salt, prodrug, metabolite or derivative thereof) and a ketone (or a salt, prodrug, metabolite or derivative thereof).

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

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

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

[0013] The present invention provides a method of therapy, the method comprising administering a combination comprising or consisting of urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0014] The present invention provides a combination for use as a medicament, the combination comprising or consisting of 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 use of a combination for the manufacture of a medicament, the combination comprising or consisting of urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0016] The present invention provides a composition comprising 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 a method for providing a health benefit to a subject, the method comprising administering to the subject a composition comprising urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0018] The present invention provides a composition for use in providing a health benefit to a subject, the composition comprising 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 use of a composition for providing a health benefit to a subject, the composition comprising urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0020] The present invention provides use of a composition for the manufacture of a medicament for providing a health benefit to a subject, the composition comprising urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0021] The present invention provides a method of therapy, the method comprising administering a composition comprising urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0022] The present invention provides a composition for use as a medicament, the composition comprising 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 use of a composition comprising 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.

[0024] The present invention provides a kit comprising a composition comprising a ketone body (or a salt, prodrug, metabolite, or derivative thereof) and a separate composition comprising urolithin (or a salt, prodrug, metabolite, or derivative thereof). DETAILED DESCRIPTION

[0025] The present invention provides a combination comprising or consisting of urolithin (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof).

[0026] Ketone body

[0027] Ketone bodies are a class of carbonyl-containing compounds produced by the liver from fatty acids under certain conditions, including acetoacetate, beta-hydroxybutyrate, and acetone. Ketone bodies are produced when fatty acid levels in the body are elevated and are metabolized for energy by the body.

[0028] Preferably, the ketone body used in the present invention is selected from the group consisting of acetoacetate or a salt, prodrug, metabolite, or derivative thereof; beta-hydroxybutyrate or a salt, prodrug, metabolite, or derivative thereof (e.g., (R)-beta-hydroxybutyrate or a salt, prodrug, metabolite, or derivative thereof or (S)-beta-hydroxybutyrate or a salt, prodrug, metabolite, or derivative thereof); and acetone or a salt, prodrug, metabolite, or derivative thereof or mixtures thereof. The ketone body is selected from the group consisting of beta-ketopentanoic acid and beta-hydroxyvaleric acid or a salt, prodrug, metabolite, or derivative thereof.

[0029] In some combinations and compositions of the present invention, the ketone body is acetoacetate or a salt, prodrug, metabolite, or derivative thereof or mixtures thereof. In some embodiments of the present invention, the ketone body is beta-hydroxybutyrate or a salt, prodrug, metabolite, or derivative thereof or mixtures thereof. In some embodiments of the present invention, the ketone body is acetone or a salt, prodrug, metabolite, or derivative thereof or mixtures thereof.

[0030] In some combinations and compositions of the present invention, the ketone body is acetoacetate. In some combinations and compositions of the present invention, the ketone body is beta-hydroxybutyrate. In some combinations and compositions of the present invention, the ketone body is acetone.

[0031] In some embodiments of the application, the combination or composition can include more than one ketone body (or salt, prodrug, metabolite, or derivative thereof). In some embodiments of the application, the combination and composition comprises acetoacetate and beta-hydroxybutyrate. In some embodiments of the application, the combination and composition comprises acetoacetate and acetone. In some embodiments of the application, the combination and composition comprises beta-hydroxybutyrate and acetone. In some embodiments of the application, the combination and composition comprises acetoacetate, beta-hydroxybutyrate, and acetone.

[0032] The present application also encompasses the use of suitable salts (e.g., pharmaceutically acceptable salts) of ketone bodies. Suitable salts according to the present application include those formed from organic or inorganic bases. Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts, such as potassium and sodium; alkaline earth metal salts, such as calcium and magnesium; 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 dimethyl-propylamine; or mono-, di- or trihydroxy lower alkylamines, such as mono-, di- or triethanolamine.

[0033] The present application also encompasses the use of prodrugs and derivatives of ketone bodies. A ketone body derivative or ketone body prodrug is a compound or substance that can be converted or metabolized into a ketone body. Suitable ketone body derivatives and ketone body prodrugs include, for example, ketone body esters (e.g., [(3R)-3-hydroxybutyl](3S)-3-hydroxybutyrate, [(3S)-3-hydroxybutyl](3R)-3-hydroxybutyrate, [(3S)-3-hydroxybutyl](3S)-3-hydroxybutyrate, or esters formed using butanediol (particularly butane-1,3-diol), arabinose, arabitol, dextrose, erythrose, fructose, galactose, glucose, glycerol, gulose, idose, lactose, lyxose, mannose, ribitol, ribose, ribulose, sucrose, talose, threose, xylitol, xylose, galactosamine, glucosamine, mannosamine, N-acetylglucosamine, mannitol, sorbitol, threitol, and (S)-1,2-propanediol), oligomers of ketone bodies or esters thereof, (R)-1,3-butanediol, triacetin, free fatty acids, and triglycerides. A preferred ketone body derivative and ketone body prodrug is beta-hydroxybutyrate-(R)-1,3-butanediol monoester.

[0034] The present application also encompasses the use of ketone body metabolites.

[0035] Those skilled in the art of organic chemistry will appreciate that many organic compounds can exist in solvated forms, in which solvents such as water, methanol, ethanol, and acetic acid are incorporated in the molecular structure of the compound. Hydrates are a form of solvate. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present application. Moreover, those skilled in the art will appreciate that the compounds of the application can exist in zwitterionic forms depending on the conditions which they are prepared, and that these forms are also encompassed within the scope of the present application. Further, those skilled in the art will appreciate that the compounds of the application can exist in different polymorphic forms or crystal forms, and that these forms are also encompassed within the scope of the present application. Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates". Those skilled in the art will appreciate that the application also encompasses solvates of the ketone bodies as well as solvates of salts, prodrugs, metabolites and derivatives thereof. Solvates include those in which the relevant solvent is pharmaceutically acceptable. Hydrates, in which the relevant solvent is water, are an example of solvates.

[0036] Urolithins:

[0037] Urolithins are metabolites of ellagitannins and ellagic acid produced by the action of the gut microbiota in mammals, including humans. Ellagitannins and ellagic acid are compounds commonly found in foods such as pomegranates, nuts and berries. Ellagitannins are minimally absorbed by the gut. Urolithins are a class of compounds having a representative formula (I) shown below. Some particularly common urolithins are described in the table below, with reference to formula (I).

[0038]

[0039] wherein A, B, C, D, W, X, Y and Z are each independently selected from H and OH.

[0040]

[0041] In practice, for commercial scale products, it is convenient to synthesize urolithins. Synthetic routes are described in, for example, WO2014 / 004902 and WO2019 / 0168972.

[0042] Particularly suitable compounds for the present application 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, and A and B are both H, and C, D and Z are all OH, then the compound is urolithin C. When W, X and Y are all H, and A, B and C are all H, and D and Z are both OH, then the compound is urolithin A. Preferably, the urolithin used in the formulations of the present application is urolithin A, urolithin B, urolithin C and / or urolithin D. More preferably, the urolithin used in the formulations of the present application is urolithin A or urolithin B. Most preferably, the urolithin used in the formulations of the present application is urolithin A.

[0043]

[0044] Iso-urolithins A and B can also be mentioned. When W, X and Y are all H, and A, B and D are all H, and C and Z are both OH, then the compound is iso-urolithin A. When W, X, Y and Z are all H, and A, B and D are all H, and C is OH, then the compound is iso-urolithin B.

[0045] The present application also encompasses the use of suitable salts (e.g. pharmaceutically acceptable salts) of urolithin. Suitable salts according to the present application include those formed from organic or inorganic bases. Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts, such as potassium and sodium; alkaline earth metal salts, such as calcium and magnesium; 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 dimethyl-propylamine; or mono-, di- or trihydroxy lower alkylamines, such as mono-, di- or triethanolamine.

[0046] The present application also encompasses the use of prodrugs of urolithin (e.g. amino acid derivatives of urolithin). One group of preferred urolithin prodrugs for use in the present application are compounds of formula (II)

[0047]

[0048] wherein A, B, C, D, W, X, Y and Z are each independently selected from H and OR 1 , provided that at least one of A, B, C, D, W, X, Y and Z is OR 1 ;

[0049] each R 1 is independently H or C(=O)R 2 , and at least one R 1 group is C(=O)R 2 ;

[0050] each R 2 is selected from:

[0051] CHR 3 NHR 4 , wherein 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,

[0052] wherein said R 3 group can optionally be substituted by one or more groups selected from halo, cyano, nitro, OR A or C 1- C4alkyl;

[0053] or R 3 and R 4together with the C and N atoms to which they are attached form a 5-membered heteroalkyl ring,

[0054] wherein the heteroalkyl ring can be optionally substituted with one or more groups selected from halogen, cyano, nitro, OR A or C 1- C3alkyl,

[0055] wherein R A is C 1- C4alkyl optionally substituted with one or more halogen, cyano or nitro. Compounds of formula (II) are disclosed in WO2015 / 097231.

[0056] In embodiments of the application, urolithin is administered as a metabolite of 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.

[0057] Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents, they are reacted in solvents or they are precipitated or crystallized from solvents. These complexes are known as “solvates”. Those skilled in the art will appreciate that the application also encompasses solvates of urolithin and solvates of salts, prodrugs, metabolites and derivatives thereof. Solvates include those in which the relevant solvent is pharmaceutically acceptable. Hydrates, in which the relevant solvent is water, are examples of solvates.

[0058] Preferably, the urolithin (or salt, prodrug, metabolite or derivative thereof) used in the compositions of the application is micronized. It has been found that micronized urolithin can dissolve or suspend faster and more efficiently than non-micronized urolithin. The micronized urolithin (or salt, prodrug, metabolite or derivative thereof) preferably has a D 50 size - that is, 50% by mass of the urolithin (or salt, prodrug, metabolite or derivative thereof) has a particle size size of less than 100 pm. More preferably, the urolithin (or salt, prodrug, metabolite or derivative thereof) has a D 50 size of less than 75 pm, for example less than 50 pm, for example less than 25 pm, for example less than 20 pm, for example less than 10 pm. More preferably, the urolithin (or salt, prodrug, metabolite or derivative thereof) has a D 50 size in the range 0.5-50 pm, for example 0.5 pm to 20 pm, for example 0.5 pm to 10 pm, for example 1 pm to 10 pm. Preferably, the urolithin (or salt, prodrug, metabolite or derivative thereof) has a D 90Size - that is, 90% of the urolithin (or salt, prodrug, metabolite or derivative thereof) by mass has a particle size of less than 100 pm. More preferably, the urolithin (or salt, prodrug, metabolite or derivative thereof) has a D50 of less than 75 pm, for example less than 50 pm, for example less than 25 pm, for example less than 20 pm, for example less than 15 pm. 90 Size. The urolithin (or salt, prodrug, metabolite or derivative thereof) preferably has a D50 in the range 5 pm to 100 pm, for example 5 pm to 50 pm, for example 5 pm to 20 pm. Micronisation can be achieved by methods established in the art, for example compressed force milling, hammer milling, general or pin milling, or jet milling (for example spiral jet milling or fluid bed jet milling) can be used. Jet milling is particularly suitable. 90 Size. The urolithin (or salt, prodrug, metabolite or derivative thereof) preferably has a D50 in the range 5 pm to 100 pm, for example 5 pm to 50 pm, for example 5 pm to 20 pm. Micronisation can be achieved by methods established in the art, for example compressed force milling, hammer milling, general or pin milling, or jet milling (for example spiral jet milling or fluid bed jet milling) can be used. Jet milling is particularly suitable.

[0059] In some embodiments, the present application relates 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 application.

[0060] In some embodiments, the present application relates to a combination or composition of the present application for use in providing a beneficial health effect to a subject.

[0061] In some embodiments, the present application relates to use of a combination or composition of the present application for providing a beneficial health effect to a subject.

[0062] In some embodiments, the present application relates to use of a combination or composition of the present application for the manufacture of a medicament for providing a beneficial health effect to a subject.

[0063] In some embodiments, the present application relates to a method of therapy, the method comprising administering to a subject a combination or composition of the present application.

[0064] In some embodiments, the present application relates to a combination or composition of the present application for use as a medicament.

[0065] In some embodiments, the present application relates to use of a combination or composition of the present application for the manufacture of a medicament.

[0066] In some embodiments, the methods, medicaments or combinations of the present application reduce muscle glycogen and / or protein breakdown during exercise or aid muscle recovery after exercise. In some embodiments, the subject is a healthy human. In other embodiments, the subject has a muscle wasting condition.

[0067] In some embodiments, the methods, medicaments or combinations of the present application reduce muscle wasting in a subject susceptible to muscle wasting. In some embodiments, the subject is a healthy human. In other embodiments, the subject has a muscle wasting condition.

[0068] In some embodiments, the reduction in muscle glycogen breakdown during exercise can be determined by the loss of glycogen in the subject. In some embodiments, the loss of glycogen in the subject is reduced by at least 10% after administration of the ketone body or ketone body ester compared to the loss of glycogen in the subject that has not been administered a ketone body or ketone body ester.

[0069] In some embodiments, the reduction in muscle glycogen breakdown during exercise can be determined by the loss of glycogen in the subject. In some embodiments, the loss of glycogen in the subject is reduced by at least 10% after administration of the ketone body or ketone body ester compared to the loss of glycogen in the subject that has not been administered a ketone body or ketone body ester.

[0070] Alternatively, the reduction in muscle glycogen and / or protein breakdown during exercise can be determined by the rate of loss of glycogen or protein. In some embodiments, the rate of loss of glycogen or protein during exercise is reduced compared to the glycogen levels in the absence of administration of a ketone body or ketone body ester.

[0071] In some embodiments, the methods, medicaments, or combinations of the present application reduce glycogen and / or protein breakdown in the subject.

[0072] In some embodiments, the methods, medicaments, or combinations of the present application reduce muscle breakdown or delay muscle wasting in a subject susceptible to muscle breakdown or muscle wasting. In some embodiments, the amount of reduction in muscle breakdown is at least 50% less than the amount of reduction in muscle breakdown in the same subject under comparable conditions not using the methods, medicaments, or combinations of the present application.

[0073] In some embodiments, the methods, medicaments, or combinations of the present application maintain or improve muscle force output in the subject. In some embodiments, the methods, medicaments, or combinations of the present application maintain or improve skeletal muscle force output. In some embodiments, the methods, medicaments, or combinations of the present application maintain or improve force output in other muscle types, such as cardiac muscle.

[0074] In some embodiments, the methods, medicaments, or combinations of the present application maintain or improve muscle force output in the subject, wherein the subject is a healthy human.

[0075] In some embodiments, the methods, medicaments, or combinations of the present application improve muscle force output in the subject, wherein the improved force output is at least 0.25% relative to placebo when measured in a 30 minute control test.

[0076] In some embodiments, the methods, medicaments, or combinations of the invention improve muscle force output in a subject, wherein the increased force output relative to placebo is at least 1 Watt when measured in a 30-minute controlled test.

[0077] In some embodiments, the methods, drugs or combinations of the present invention can be used to improve muscle performance, maintain or improve muscle function, prevent muscle function decline, increase muscle mass and / or reduce muscle wasting. Improving muscle performance, improving or maintaining muscle function, increasing muscle mass and / or reducing muscle wasting can be part of a medical treatment, or can be for personal preference ("lifestyle") or cosmetic reasons. The compositions of the present invention can also be used to reduce muscle inflammation after exercise and / or enhance muscle recovery after exercise.

[0078] The method, medicament or combination of the present invention can be used as a medicament. The method, medicament or combination can be used as a dietary supplement, as a functional food, as a functional drink or as a medical food.

[0079] The methods, medicaments, or compositions of the present invention can be used to enhance muscle performance. Thus, the present invention provides methods, medicaments, or compositions of the present invention for enhancing muscle performance. The present invention also provides a method for enhancing muscle performance by administering an effective amount of a composition of the present invention to a subject. Administration can be self-administered.

[0080] Enhanced muscle performance can be one or more of improved muscle function, reduced muscle function decline, improved muscle strength, improved muscle endurance, and improved muscle recovery.

[0081] Thus, the compositions of the present invention can be used in methods of improving physical endurance (e.g., the ability to perform physical tasks such as exercise, physical labor, sports activities, etc.), inhibiting or delaying physical fatigue, enhancing work capacity, increasing endurance and / or reducing muscle fatigue.

[0082] Improved muscle function may be particularly beneficial for elderly subjects whose muscle function decreases due to old age or age-related conditions. For example, subjects who may benefit from improved muscle function may have experienced a decrease in muscle function, which then leads to pre-frailty and frailty. In addition to their decreased muscle function, such subjects do not necessarily experience muscle wasting. Some subjects do experience muscle wasting and decreased muscle function, such as subjects with sarcopenia. The compositions of the present invention can be used to enhance muscle performance by administering the compositions of the present invention to frail or pre-frail subjects.

[0083] In further embodiments, in the method of enhancing muscle performance, the subject has age-related decline in muscle function, age-related sarcopenia, age-related muscle wasting, physical fatigue, muscle fatigue, and / or is frail or pre-frail. In further embodiments of the method of enhancing muscle performance, the subject suffers from physical fatigue or muscle fatigue, and / or wherein the subject is frail or pre-frail.

[0084] Muscle performance can be athletic performance, that is, the ability of an athlete to perform muscle performance when participating in a physical activity. Enhancement of athletic performance, strength, speed, and endurance is measured by an increase in muscle contraction strength, an increase in muscle contraction amplitude, or a decrease in muscle reaction time between stimulation and contraction. Athletes refer to individuals who participate in sports at any level and seek to improve the level of strength, speed, or endurance in their performance, for example, body builders, cyclists, long-distance runners, and sprinters. Enhanced athletic performance is manifested in the ability to overcome muscle fatigue, the ability to maintain activity over long periods, and more effective training.

[0085] The methods, medicaments, or combinations of the present application can improve physical performance in individuals with a disease, including young and old individuals. The compositions of the present application can improve physical performance, for example, short-term performance or long-term performance in healthy individuals, including athletes, non-athletic individuals, sedentary individuals, and the elderly. This improvement in performance can be measured by the time taken to walk or run a specific distance, for example, improved performance during a 6-minute walk test (MWT), improvement in the time to run a specific distance, improvement in the IPAQ score on the International Physical Activity Questionnaire, increase in the number of times standing from a chair in a specific time, or another test designed to measure physical performance.

[0086] According to further embodiments of the present application, there is provided a composition of the present application for use in a method of enhancing physical performance in a subject, for example, an elite athlete or a sub-elite athlete.

[0087] In certain embodiments, enhancing physical performance comprises at least one effect selected from the group consisting of: enhancing athletic performance, enhancing running performance, enhancing muscle performance, enhancing aerobic endurance, enhancing perceived exertion level, reducing post-exercise fatigue, enhancing muscle recovery, reducing exercise-induced muscle damage, reducing muscle soreness, and enhancing exercise-induced muscle damage repair. In further embodiments, enhancing physical performance comprises enhancing muscle performance during high intensity aerobic activity. In still further embodiments, enhancing physical performance comprises improving aerobic endurance during high intensity aerobic activity. In still further embodiments, enhancing physical performance comprises improving resting metabolic rate (RMR). In certain embodiments, enhancing physical performance results in improved athletic performance. In further embodiments, enhancing physical performance results in improved race completion time. In still further embodiments, enhancing physical performance results in reduced perceived exertion level (RPE).

[0088] In one embodiment, enhancing physical performance comprises enhancing muscle performance during high intensity aerobic activity.

[0089] In one embodiment, enhancing physical performance comprises improving aerobic endurance during high intensity aerobic activity.

[0090] In one embodiment, enhancing physical performance comprises improving resting metabolic rate (RMR).

[0091] In one embodiment, enhancing physical performance results in improved athletic performance.

[0092] In one embodiment, enhancing physical performance results in improved race completion time.

[0093] In one embodiment, enhancing physical performance results in reduced perceived exertion level (RPE).

[0094] According to further embodiments of the present application, there is provided a composition of the present application for use in a method of enhancing physical recovery in a subject, e.g., an elite athlete or a sub-elite athlete.

[0095] In certain embodiments, the enhancement of physical recovery after high intensity aerobic activity. In further embodiments, the enhancing physical recovery comprises at least one effect selected from the group consisting of: enhancing muscle recovery, enhancing athletic performance, enhancing running performance, enhancing muscle performance, enhancing aerobic endurance, enhancing self-perceived exertion, reducing post-exercise fatigue, enhancing muscle recovery, reducing exercise-induced muscle damage, reducing muscle soreness, and enhancing exercise-induced muscle damage repair. In still further embodiments, the enhancing physical recovery comprises enhancing muscle recovery after high intensity aerobic activity. In still further embodiments, the enhancing physical recovery comprises reducing muscle soreness after high intensity aerobic activity. In certain embodiments, the enhancing physical recovery comprises reducing creatine kinase (CK) levels in the subject after aerobic activity (as compared to baseline), as measured by the area under the plasma concentration-time curve (AUC CK ) of CK. CRP ) of C-reactive protein (CRP).

[0096] In one embodiment, the enhancement of physical recovery is after high intensity aerobic exercise.

[0097] In one embodiment, the enhancing physical recovery comprises enhancing muscle recovery after high intensity aerobic activity.

[0098] In one embodiment, the enhancing physical recovery comprises reducing muscle soreness after high intensity aerobic activity.

[0099] In one embodiment, the enhancing physical recovery comprises reducing creatine kinase (CK) levels in the subject after aerobic activity (as compared to baseline), as measured by the area under the plasma concentration-time curve (AUC CK ) of CK.

[0100] In one embodiment, the enhancing recovery comprises reducing C-reactive protein (CRP) levels in the subject after aerobic activity (as compared to baseline), as measured by the area under the plasma concentration-time curve (AUC CRP ) of CRP.

[0101] According to further embodiments of the application, there is provided the composition of the application for use in a method of enhancing endurance in a subject (e.g., elite athlete or sub-elite athlete).

[0102] The compositions of the present invention further provide for improvements in the composition of the compositions. Endurance refers to the time to reach fatigue when exercising at a constant work load (typically intensity <80% VO2max). The compositions of the present invention can improve endurance in individuals with disease, including young and older individuals. The compositions of the present invention can improve endurance in healthy individuals, including athletes, non-athletic individuals, sedentary individuals, and older adults. The present invention provides a method to increase the time to reach fatigue when performing a specific activity (e.g., weight training, walking, running, swimming, or cycling). This improvement in endurance can be assessed by objective measures (e.g., speed, oxygen consumption, or heart rate) or can be assessed by self-reported measures (e.g., using validated questionnaires).

[0103] In certain embodiments, the methods comprise enhancing physical endurance during high intensity aerobic activity. In further embodiments, enhancing physical endurance comprises at least one effect selected from the group consisting of: enhancing muscle recovery, enhancing athletic performance, enhancing running performance, enhancing muscle performance, enhancing aerobic endurance, enhancing perceived exertion, reducing post-exercise fatigue, enhancing muscle recovery, reducing exercise-induced muscle damage, reducing muscle soreness, and enhancing exercise-induced muscle damage repair. In yet further embodiments, enhancing physical endurance comprises enhancing muscle endurance. In still further embodiments, enhancing physical endurance comprises increasing the maximal oxygen consumption (VO 2max ) of the subject.

[0104] In one embodiment, the enhancement in physical recovery is during high intensity aerobic exercise.

[0105] In one embodiment, enhancing physical endurance comprises enhancing muscle endurance.

[0106] In one embodiment, enhancing physical endurance comprises increasing the maximal oxygen consumption (VO2max) of the subject.

[0107] In certain embodiments, the methods of the present disclosure comprise administering to a subject in need thereof an effective amount of a composition of the present invention, 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 VO 2max max greater than 65 mL kg-1min-1. In further embodiments, the subject has a 3 km run personal best of less than 9 minutes. In other embodiments, the subject is a sub-elite athlete. In certain such embodiments, the subject has a VO 2maxIn further embodiments, the subject has a 3 km run personal best of about 9 minutes to about 10 minutes. In certain embodiments, the subject is about 18 years old to about 45 years old.

[0108] In some embodiments, the methods, medicaments, or combinations of the present application are used to reduce inflammation and / or improve resistance to inflammation.

[0109] In some embodiments, the methods, medicaments, or combinations of the present application are used to treat cognitive dysfunction. In some embodiments, the methods, medicaments, or combinations of the present application improve cardiac efficiency. In some embodiments, the methods, medicaments, or combinations of the present application improve brain metabolic efficiency. In some embodiments, the methods, medicaments, or combinations of the present application reduce the effects of neurodegenerative disease.

[0110] In some embodiments, the methods, medicaments, or combinations of the present application are used to treat Alzheimer’s disease or Parkinson’s disease.

[0111] In some embodiments, the methods, medicaments, or combinations of the present application are used to treat muscle injury or fatigue.

[0112] In some embodiments, the methods, medicaments, or combinations of the present application maintain or improve cognitive function under fatigue or reduce the adverse effects on cognitive function under fatigue.

[0113] Dosage:

[0114] The effective amount of the combination components to be taken will vary depending on the mode of administration, age, body weight, and general health of the subject. Factors such as the subject’s disease state, age, and body weight can be important and dosage regimens can be adjusted to provide an optimum response.

[0115] The daily dosage of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) provided by the compositions of the present application can range from 20 mg to 5000 mg, such as 20 mg to 4000 mg, such as 200 mg to 4000 mg, such as 20 mg to 3000 mg, such as 20 mg to 2000 mg, such as 100 mg to 1000 mg, such as 100 mg to 800 mg, such as 200 mg to 600 mg, such as 200 mg to 400 mg, such as 200 mg to 300 mg, such as 250 mg. The daily intake of ketone bodies (or salts, prodrugs, metabolites, or derivatives thereof) can be provided as a single portion or can be divided into multiple portions.

[0116] Alternatively, the compositions of the present application can provide a daily dose of ketone bodies (or salts, prodrugs, metabolites or derivatives thereof) in the range of 2g to 50g, for example 5g to 30g, for example 10g to 20g, for example 2g, for example 3g, for example 4g, for example 5g, for example 6g, for example 7g, for example 8g, for example 9g, for example 10g, for example 1 lg, for example 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, for example 21g, for example 22g, for example 23g, for example 24g, for example 25g, for example 26g, for example 27g, for example 28g, for example 29g, for example 30g, for example 31g, for example 32g, for example 33g, for example 34g, for example 35g, for example 36g, for example 37g, for example 38g, for example 39g, for example 40g, for example 41g, for example 42g, for example 43g, for example 44g, for example 45g, for example 46g, for example 47g, for example 48g, for example 49g, for example 50g. The daily intake of ketone bodies (or salts, prodrugs, metabolites or derivatives thereof) can be provided as a single portion or can be divided into multiple portions.

[0117] The unit dose can be in the form of a snack bar; a snack bar having a weight of 25g to 150g, for example 40g to 100g. The snack bar can contain the required amount of ketone bodies (or salts, prodrugs, metabolites or derivatives thereof) (such as 200mg to 300mg ketone bodies (or salts, prodrugs, metabolites or derivatives thereof) or other amounts mentioned above). Alternatively, the unit dose composition can be in the form of a beverage, for example provided in a container (such as a sachet or bottle) suitable for a single dose (for example 50mL to 500mL, for example 100mL to 300mL). A 100mL to 300mL beverage can contain the required amount of ketone bodies (or salts, prodrugs, metabolites or derivatives thereof). Alternatively, the unit dose composition can be in the form of a powder to be reconstituted into a beverage, for example an appropriate amount of powder for a single dose (for example 5g to 10g powder containing 200mg to 300mg ketone bodies (or salts, prodrugs, metabolites or derivatives thereof)). A 100mL to 500mL reconstituted beverage can contain the required amount of ketone bodies (or salts, prodrugs, metabolites or derivatives thereof). As mentioned below, the compositions for use in the present application can take any suitable physical form. The compositions can be in the form of a solid (such as a tablet or bar), a semi-solid (such as a soft capsule, a capsule (such as a hard capsule) or a dragee), a powder or a liquid (including an emulsion). The compositions of the present application can be a nutritional composition. The compositions of the present application can be a pharmaceutical composition. The compositions can be in the form of a dietary supplement, as a functional food, a functional beverage or as a medical food or a medical nutritional product.

[0118] The daily intake of the urolithin (or salt, prodrug, metabolite, or derivative thereof, e.g., urolithin A) component is typically in the range of 10 mg to 5 g per day, e.g., 20 mg to 2500 mg per day, e.g., 25 mg to 250 mg, e.g., 25 mg to 500 mg, e.g., 50 mg to 1500 mg per day, e.g., 250 mg to 2000 mg, e.g., 250 mg to 1500 mg per day, e.g., 50 mg to 1000 mg per day, e.g., 20 mg to 250 mg per day, e.g., 250 mg to 1000 mg per day, e.g., 500 mg to 1000 mg per day, e.g., 750 mg to 1000 mg per day. In one embodiment, the composition is taken in an amount to provide a urolithin (or salt, prodrug, metabolite, or derivative thereof) dose in the range of about 0.2 mg / kg / day to greater than about 100 mg / kg / day.For example, the dosage of urolithin (or salt, prodrug, metabolite, or derivative thereof) can 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 to 25 mg / kg / day, 0.25 mg / kg / day to 10 mg / kg / day, 0.25 mg / kg / day to 7.5 mg / kg / day, 0.25 mg / kg / day to 5 mg / kg / day, 0.5 mg / kg / day to 50 mg / kg / day, 0.5 mg / kg / day to 25 mg / kg / day, 0.5 mg / kg / day to 20 mg / kg / day, 0.5 mg / kg / day to 15 mg / kg / day, 0.5 mg / kg / day to 10 mg / kg / day, 0.5 mg / kg / day to 7.5 mg / kg / day, 0.5 mg / kg / day to 5 mg / kg / day, 0.75 mg / kg / day to 50 mg / kg / day, 0.75 mg / kg / day to 25 mg / kg / day, 0.75 mg / kg / day to 20 mg / kg / day, 0.75 mg / kg / day to 15 mg / kg / day, 0.75 mg / kg / day to 10 mg / kg / day, 0.75 mg / kg / day to 7.5 mg / kg / day, 0.75 mg / kg / day to 5 mg / kg / day, 1.0 mg / kg / day to 50 mg / kg / day, 1 mg / kg / day to 25 mg / kg / day, 1 mg / kg / day to 20 mg / kg / day, 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.

[0119] A unit dose can be in the form of a snack bar; a snack bar having a weight of 25g to 150g (e.g. 40g to 100g) can contain the required amount of urolithin (or salt, prodrug, metabolite or derivative thereof). Alternatively, a unit dose composition can be in the form of a beverage, for example provided in a container (e.g. sachet) suitable for a single dose volume (e.g. 100mL to 300mL). A 50mL to 500mL (e.g. 100mL to 300mL) beverage can contain the required amount of urolithin (or salt, prodrug, metabolite or derivative thereof). A beverage providing a composition of the application can contain urolithin (or salt, prodrug, metabolite or derivative thereof) at a concentration of 0.1 mg / mL to 50 mg / mL, for example 0.5 mg / mL to 10 mg / mL, for example 1 mg / mL to 5 mg / mL. Alternatively, a unit dose can be in the form of one or more solids, such as a compressed tablet. A single compressed tablet can contain a dose of urolithin (or salt, prodrug, metabolite or derivative thereof), for example 25mg, 50mg, 75mg, 100mg, 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 600mg, 700mg, 800mg, 900mg or 1000mg. Alternatively, a unit dose can be in the form of one or more semi-solid doses, such as a soft gel capsule or a paste. A single soft gel capsule can contain a dose of urolithin (or salt, prodrug, metabolite or derivative thereof), for example 25mg, 50mg, 75mg, 100mg, 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg or 500mg, for example 250mg.

[0120] The weight ratio between ketone body (or salt, prodrug, metabolite or derivative thereof) and urolithin (or salt, prodrug, metabolite or derivative thereof) is typically in the range of 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 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 can 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.

[0121] Thus, the compositions of the present application can contain 20 mg to 5000 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 10 mg to 5 g of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 20 mg to 4000 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 10 mg to 5 g of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 20 mg to 3000 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 10 mg to 5 g of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 20 mg to 2000 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 10 mg to 5 g of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 150 mg to 3000 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 10 mg to 5 g of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 50 mg to 500 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 50 mg to 500 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 250 mg to 2000 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 10 mg to 3000 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 300 mg to 3000 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 10 mg to 3000 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 200 mg to 600 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 20 mg to 2500 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 250 mg to 2500 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 100 mg to 2000 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 200 mg to 400 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 20 mg to 2500 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); for example, 200 mg to 300 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 50 mg to 1000 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); 200 mg to 300 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 50 mg to 500 mg of urolithin (or salt, prodrug, metabolite or derivative thereof); 200 mg to 300 mg of ketone body (or salt, prodrug, metabolite or derivative thereof) and 100 mg to 500 mg of urolithin (or salt, prodrug, metabolite or derivative thereof). The compositions can further contain, for example, proteins, carbohydrates, vitamins, and minerals.

[0122] The dosages of the compounds mentioned herein are on a daily dose basis. In many cases, the beneficial effects of the compositions of the invention are most apparent when the compositions are taken for an extended period of time, e.g., 2 weeks or longer, e.g., 4 weeks or longer, e.g., 6 weeks or longer, e.g., 8 weeks, e.g., 12 weeks or longer, e.g., 16 weeks or longer, e.g., 20 weeks or longer, e.g., 24 weeks or longer.

[0123] In the methods of the present invention, the urolithin (or its salt, prodrug, metabolite, or derivative) and the ketone body (or its salt, prodrug, metabolite, or derivative) do not need to be administered simultaneously as part of a single composition. The urolithin (or its salt, prodrug, metabolite, or derivative) and the ketone body (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 body (or its salt, prodrug, metabolite, or derivative). Alternatively, the ketone body (or its salt, prodrug, metabolite, or derivative) can be administered first, followed by the urolithin (or its salt, prodrug, metabolite, or derivative). The present invention further provides a kit, for example, for use in the methods of the present invention, comprising the urolithin (or its salt, prodrug, metabolite, or derivative) and the ketone body (or its salt, prodrug, metabolite, or derivative). The urolithin (or its salt, prodrug, metabolite, or derivative) and the ketone body (or its salt, prodrug, metabolite, or derivative) can be in different physical forms.

[0124] Preferably, the urolithin (or its salt, prodrug, metabolite or derivative) and the ketone body (or its salt, prodrug, metabolite or derivative) are administered simultaneously as part of a single composition. Thus, the present invention also provides a composition comprising a urolithin (or its salt, prodrug, metabolite or derivative) and a ketone body (or its salt, prodrug, metabolite or derivative). The composition of the present invention can be used to replace the combination of the present invention in all embodiments described herein. In addition, all disclosures (e.g., dosages) relating to urolithins (or their salts, prodrugs, metabolites or derivatives) and ketone bodies (or their salts, prodrugs, metabolites or derivatives), and combinations thereof, apply to the combinations of the present invention and the compositions of the present invention.

[0125] In some embodiments, the compositions of the present invention comprise a naturally occurring urolithin (or a salt, prodrug, metabolite, or derivative thereof). In some embodiments, the compositions of the present invention comprise a compound of formula (I) or a salt, prodrug, metabolite, or derivative thereof. In some embodiments, the compositions of the present invention comprise a compound of formula (I) selected from a compound of formula (I) or a salt, prodrug, metabolite, or derivative thereof, wherein:

[0126] -Z is OH and W, X and Y are all H;

[0127] - A, B, W, X, and Y are all H, and C, D, and Z are all OH (i.e., urolithin C);

[0128] - A, B, C, W, X, and Y are all H, and D and Z are both OH (i.e., urolithin A).

[0129] In some embodiments, the compositions of the present application comprise urolithin A, urolithin B, urolithin C, and / or urolithin D, or salts, prodrugs, metabolites, or derivatives thereof.

[0130] In some embodiments, the compositions of the present application comprise urolithin A, or salts, prodrugs, metabolites, or derivatives thereof.

[0131] In some embodiments, the compositions of the present application comprise urolithin A.

[0132] In some embodiments, the compositions of the present application comprise two or more urolithins (or salts, prodrugs, metabolites, or derivatives thereof).

[0133] In some embodiments, the compositions of the present application comprise a naturally occurring ketone (or salts, prodrugs, metabolites, or derivatives thereof). In some embodiments, the compositions of the present application comprise acetoacetate or salts, prodrugs, metabolites, or derivatives thereof. In some embodiments, the compositions of the present application comprise beta-hydroxybutyrate or salts, prodrugs, metabolites, or derivatives thereof. In some embodiments, the compositions of the present application comprise acetone or salts, prodrugs, metabolites, or derivatives thereof. In some embodiments, the compositions of the present application comprise acetoacetate (or salts, prodrugs, metabolites, or derivatives thereof) and beta-hydroxybutyrate (or salts, prodrugs, metabolites, or derivatives thereof). In some embodiments, the compositions of the present application comprise acetoacetate (or salts, prodrugs, metabolites, or derivatives thereof) and acetone (or salts, prodrugs, metabolites, or derivatives thereof). In some embodiments, the compositions of the present application comprise beta-hydroxybutyrate (or salts, prodrugs, metabolites, or derivatives thereof) and acetone (or salts, prodrugs, metabolites, or derivatives thereof). In some embodiments, the compositions of the present application comprise acetoacetate (or salts, prodrugs, metabolites, or derivatives thereof), beta-hydroxybutyrate (or salts, prodrugs, metabolites, or derivatives thereof), and acetone (or salts, prodrugs, metabolites, or derivatives thereof). In some embodiments, the compositions of the present application comprise acetoacetate. In some embodiments, the compositions of the present application comprise beta-hydroxybutyrate. In some embodiments, the compositions of the present application comprise acetone. In some embodiments, the compositions of the present application comprise acetoacetate and beta-hydroxybutyrate. In some embodiments, the compositions of the present application comprise acetoacetate and acetone. In some embodiments, the compositions of the present application comprise beta-hydroxybutyrate and acetone. In some embodiments, the compositions of the present application comprise acetoacetate, beta-hydroxybutyrate, and acetone.

[0134] In some embodiments, the compositions of the present application comprise more than one ketone body (or salt, prodrug, metabolite or derivative thereof).

[0135] Form of composition:

[0136] The compositions of the present application can take any suitable physical form. The compositions can be in the form of a solid (e.g. tablet or bar), semi-solid (e.g. soft gelatin capsule, capsule (e.g. hard gelatin capsule) or dragee), powder or liquid (including emulsion). The compositions of the present application can be a nutritional composition. The compositions of the present application can be a pharmaceutical composition. The compositions can be in the form of a dietary supplement, as a functional food, functional beverage or as a medical food or medical nutritional product.

[0137] The compositions in the form of tablets can be of any suitable type and they can contain excipients that are conventional in the art. For example, the excipients can provide the desired hardness, shelf life and flavour, so that the composition has an acceptable taste, an attractive appearance and good storage stability. The bars can be of any suitable type and they can contain ingredients that are conventionally used to make snack bars.

[0138] The snack bars can be unit doses of the compositions of the present application; snack bars having a weight of 25g to 150g, for example 40g to 100g, can contain the required amount of ketone body (or salt, prodrug, metabolite or derivative thereof) as mentioned above as well as the required amount of urolithin (or salt, prodrug, metabolite or derivative thereof) as mentioned above.

[0139] The semi-solid forms can likewise contain excipients that are conventional in the art. For example, the excipients can provide the desired hardness, shelf life and flavour, so that the composition has an acceptable taste, an attractive appearance and good storage stability. The semi-solid forms can be provided for oral administration or for topical administration.

[0140] Powders are often used to supply nutritional and pharmaceutical compositions. Powders have the advantage that multiple doses can be provided in a single container, and doses of various sizes can be used from the same supply container. Powders generally have good storage characteristics. Powder compositions can also contain excipients that are conventional in the art. For example, excipients can provide shelf life, flavor, and moisture resistance, so that the composition has an acceptable taste, an appealing appearance, and good storage stability. The present invention can take the form of a kit comprising a composition comprising a ketone body (or salt, prodrug, metabolite, or derivative thereof) and a separate composition comprising a urolithin (or salt, prodrug, metabolite, or derivative thereof), for example a ketone body powder composition and a separate solid or liquid composition containing a urolithin (or salt, prodrug, metabolite, or derivative thereof). A solid or liquid composition containing a urolithin (or salt, prodrug, metabolite, or derivative thereof), for example a tablet or beverage, or other form described herein, can be provided with instructions for use and a ketone powder. For example, both the ketone body (or salt, prodrug, metabolite, or derivative thereof) and the urolithin (or salt, prodrug, metabolite, or derivative thereof) can be in powder form.

[0141] Liquid compositions can be in the form of a medicament, a beverage. Liquid formulations can be a solution, an emulsion, a slurry, or other semi-liquid. For example, excipients in a liquid composition can provide shelf life, visual appearance, flavor, and mouthfeel, so that the composition has an acceptable taste, an appealing appearance, and good storage stability. Liquid compositions can be provided for oral administration. Liquid compositions can be provided for topical application, for example in the form of a cream, ointment, or lotion.

[0142] A beverage can be a unit dose of a composition of the present invention, for example 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, for example 100 mL to 300 mL). A 100 mL to 300 mL beverage can contain a ketone body (or salt, prodrug, metabolite, or derivative thereof) in the required amount as mentioned above and a urolithin (or salt, prodrug, metabolite, or derivative thereof) in the required amount as mentioned above.

[0143] Alternatively, a powder to be reconstituted into a beverage can be a unit dose of a composition of the present invention, for example a suitable amount of powder for a single dose (e.g. 5 g to 10 g of powder containing 200 mg to 300 mg of a ketone body (or salt, prodrug, metabolite, or derivative thereof) and). A 100 mL to 500 mL reconstituted beverage can contain a ketone body (or salt, prodrug, metabolite, or derivative thereof) in the required amount as mentioned above and a urolithin (or salt, prodrug, metabolite, or derivative thereof) in the required amount as mentioned above.

[0144] For some uses, the compositions of the present application can also be in the form of a solution suitable for injection or intravenous administration.

[0145] Additional components in the compositions of the present application:

[0146] The compositions according to the present application can contain additional components other than urolithin (or salts, prodrugs, metabolites or derivatives thereof) and ketone body (or salts, prodrugs, metabolites or derivatives thereof). The additional components can 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.

[0147] European Pharmacopoeia describes medium chain triglycerides as fixed oils extracted from the hard dry part of the endosperm of Cocos nucifera L. (coconut) or from the dried endosperm of Elaeis guineenis Jacq. (African oil palm). Both the European Pharmacopoeia and the USP NF have specifications for medium chain triglycerides which require the presence of specific 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%.

[0148] The medium chain triglycerides for use in the preferred compositions comprise a mixture of triglycerides with fatty acid chains present in the following proportions: 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%.

[0149] The medium chain triglycerides for use in the preferred compositions can be from any known or otherwise suitable source.

[0150] Among the vitamins, mention can be made in particular of vitamin A, vitamin C, vitamin D, vitamin E, vitamin B12 and vitamin K2. As used herein, “vitamin D” means any known form of vitamin D and includes in particular vitamin D2 (ergocalciferol), vitamin D3 (cholecalciferol), precursors, metabolites and other analogues of vitamin D and combinations thereof, as well as various active and inactive forms of vitamin D. For example, vitamin D3 can be provided in its non-hydroxylated, inactive form (e.g. cholecalciferol) or can be provided in its hydroxylated, active form (e.g. calcitriol).

[0151] Creatine has been described as having beneficial effects in the treatment of muscle disorders. The creatine can be included in the compositions of the present application. β-hydroxy-β-methylbutyrate (HMB) has been described as having beneficial effects in the treatment of muscle disorders. It can be included in the compositions of the present application.

[0152] Among the minerals, mention can be made in particular of calcium salts (for example calcium phosphate), selenium, zinc salts, magnesium salts and iron salts.

[0153] Polyunsaturated fatty acids are fatty acids containing more than one double bond in the main chain. These include many important compounds, such as essential fatty acids, for example omega-3 and omega-6 fatty acids. Long-chain polyunsaturated fatty acids are suitable, and are preferably fatty acids containing at least 20 carbon atoms in the molecule. 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-5,8,11,14,17-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-tetracosa- pentaenoic acid C24:5; cis-6,9,12,15,18,21-tetracosahexaenoic acid (gadoleic 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, cis-4,7,10,13,16-docosapentaenoic acid (Osbond acid) C22:5. The compositions according to the application preferably contain EPA, DHA or a combination thereof, for example from 10 mg to 1000 mg per serving; for example from 25 mg to 250 mg per serving. It is beneficial for many muscle growth and / or muscle strengthening therapies 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 referred to as "functional amino acids". The compositions of the present application can include one or more branched-chain amino acids (leucine, isoleucine and valine). The compositions of the present application can include one or both of L-arginine and L-glutamine. The compositions of the present application can include lysine.

[0154] The pharmaceutical compositions of the present invention can include additional pharmaceutically active compounds. For example, a statin can be included. The present invention can be provided as a kit comprising a composition of urapidil (or a salt, prodrug, metabolite, or derivative thereof) and a ketone body (or a salt, prodrug, metabolite, or derivative thereof); and a pharmaceutically active compound, such as a statin.

[0155] The compositions of the present invention can include one or more additional agents useful for mitochondrial biosynthesis or treatment of mitochondrial disorders. Such compounds include, but are not limited to, resveratrol, pyrroloquinoline quinone, ubiquinone, sulforaphane, coenzyme Q10, genistein, hydroxytyrosol, quercetin, L-carnitine, alpha-lipoic acid, and folate (e.g., folinic acid).

[0156] The compositions of the present invention can further (or alternatively) include additional compounds, including, for example, tomatidine, ursolic acid, curcumin, capsaicin, menthol, triethanolamine salicylate, and methyl salicylate.

[0157] The additional compound can be an additional active ingredient or supplement, such as carnitine or a salt thereof. The term carnitine encompasses L-carnitine and derivatives thereof, including acetyl-L-carnitine (ALCAR) and propionyl L-carnitine. Salts of carnitine include tartrate salts, such as in the case of L-carnitine L-tartrate (LCLT), and glycine salts, such as glycine propionyl-L-carnitine (GPLC). When used, carnitine can be administered by any suitable means or dosage form, but typically carnitine is administered orally, and dosed daily.

[0158] In some exemplary embodiments, the compositions of the present disclosure can comprise one or more additional macronutrients, typically protein, fat, or carbohydrate, or two or more of protein, fat, and carbohydrate, in addition to one or more ketone bodies (or a salt, prodrug, metabolite, or derivative thereof) and urapidil (or a salt, prodrug, metabolite, or derivative thereof).

[0159] Any suitable fat or oil source typically used in the preparation of foods and pharmaceuticals 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; rapeseed oil; marine oil; cottonseed oil; polyunsaturated fatty acids such as docosahexaenoic acid (DHA), arachidonic acid (ARA), eicosapentaenoic acid (EPA), and combinations thereof.

[0160] Non-limiting examples of carbohydrates or sources thereof suitable for use in the compositions described herein can include maltodextrin, hydrolyzed or modified starch or corn starch, glucose polymers, corn syrup, corn syrup solids, rice-derived carbohydrates, glucose, fructose, lactose, trehalose, high fructose corn syrup, tapioca dextrin, isomaltulose, sucromalt, maltitol powder, glycerol, 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.

[0161] Non-limiting examples of proteins or sources thereof suitable for use in the compositions described herein can include hydrolyzed, partially hydrolyzed, or non-hydrolyzed proteins or protein sources. They can be from any known or otherwise suitable source, such as milk (e.g., casein, whey), animal (e.g., meat, fish), cereal (e.g., rice, corn), or vegetable (e.g., soy, pea) sources. Combinations of various sources or types of proteins can 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 concentrate, whey protein isolate, sodium or calcium caseinate, whole milk, partially or completely skimmed milk, yogurt, soy protein isolate, and soy protein concentrate, and combinations thereof. Combinations of various sources or types of proteins can be used. For example, Greek and Icelandic style yogurts are known to typically have a particularly high protein content, which makes them particularly suitable for use in the formulations of the present application. Yogurts for use in the compositions of the present application can contain, for example, 2 g to 15 g protein / 100 g. Particularly preferred are yogurts with a high protein content, for example, 6 g to 15 g / 100 g, for example, 7 g to 15 g / 100 g, for example, 8 g to 15 g / 100 g. Optionally, supplemental protein can also be added to the yogurt formulation to increase the protein content in the formulation. Yogurts of the present application can contain live cultures, such as S. thermophilus, L. bulgaricus, L. acidophilus, or L. lactis.

[0162] The total concentration or amount of protein, fat, carbohydrate, and other components varies depending on the nutritional needs of the target user.

[0163] The additional components in the compositions of the present application can be compounds that do not provide a health benefit to the subject, but can improve the composition in some other way, such as the taste, texture, or shelf life thereof, mentioned above. Thus, the compositions of the present application can further contain one or more compounds selected from emulsifiers, colorants, preservatives, gums, coagulants, thickening agents, sweeteners, and flavorings.

[0164] Suitable emulsifiers, colorants, preservatives, gums, coagulants, and thickening agents are well known in the art of emulsion and other semi-liquid manufacture. For example, preservatives such as benzoic acid, sorbic acid, phosphoric acid, lactic acid, acetic acid, hydrochloric acid, and soluble salts thereof can be used.

[0165] Emulsifiers can include one or more of phosphatidylcholine, lecithin, polysorbate such as polysorbate 60 or polysorbate 80 (Tween-60 and Tween-80), and glycerol monostearate (GMS). Glycerol monostearate is also known as glycerin monostearate.

[0166] Stabilizers can be used in the compositions described herein. Many compositions are stable suspensions that do not require the addition of a stabilizer. A stable suspension is one that does not phase separate over time. For some compositions, the stability can be improved by including an added stabilizer. Suitable stabilizers for use in the compositions of the present application include glycerol monostearate (GMS), silicon dioxide, and vegetable shortening. One exemplary stabilizer is GMS and the preferred compositions of the present application contain GMS. GMS is also a good solvent for phospholipids such as those found in lecithin. GMS has two crystalline forms: the alpha form is dispersible and foamy and can be used as an emulsifier or preservative. The beta form is suitable for use in wax bases. The alpha form will convert to the beta form when heated to 50°C.

[0167] GMS is available in two different grades: 40-55% glycerol monoesters and 90% glycerol monoesters. The 40-55% glycerol monoesters as defined by the European Pharmacopoeia describe GMS as a mixture of monoacylglycerols (mainly monostearoylglycerol) with amounts of di- and triglycerols. Specifically, the 40-55 grade contains 40-55% monoacylglycerols, 30-45% diacylglycerols, and 5-15% triacylglycerols. The 99% grade contains not less than 90% glycerol monoesters. The glycerol monoesters in commercial GMS products are a mixture of glycerol monostearate and glycerol monopalmitate in varying proportions. The European Pharmacopoeia further classifies monostearoylglycerol 40-55 into three types based on the proportion of stearate esters in the mixture. Type 1 contains 40.0-60.0% stearic acid, and the sum of palmitic and stearic acids is < 90%. Type 2 contains 60.0-80.0% stearic acid, and the sum of palmitic and stearic acids is < 90%. Type 3 contains 90.0-99.0% stearic acid, and the sum of palmitic and stearic acids is < 96%. Any form of GMS can be used in the composition.

[0168] In some embodiments, the compositions of the present application comprise urolithin (e.g., urolithin A) or a salt, prodrug, metabolite, or derivative thereof; one or more ketone body or a salt, prodrug, metabolite, or derivative thereof; medium chain triglyceride; and a stabilizer, e.g., glycerol monostearate. In some embodiments, the compositions of the present application comprise urolithin (e.g., urolithin A) or a salt, prodrug, metabolite, or derivative thereof; one or more ketone body or a salt, prodrug, metabolite, or derivative thereof; medium chain triglyceride; an emulsifier; and a stabilizer.

[0169] Metal chelators or sequestering agents such as the sodium calcium salt of ethylenediaminetetraacetic acid (EDTA) can also be used in the compositions of the present application. Other components that can be included in the formulations of the present application include polyethylene glycol, silicon dioxide, vegetable shortening, and beeswax.

[0170] Sweeteners can be particularly beneficial in the compositions of the present application. High potency non-nutritive carbohydrate sweeteners can be used, e.g., selected from the group consisting of aspartame, sucralose, acesulfame potassium, saccharin, cyclamate, stevia, thaumatin, and mixtures thereof. Aspartame is particularly suitable.

[0171] Flavorings can be particularly beneficial for the compositions of the present application. In liquid or semi-liquid compositions, fruit flavors can be provided by including a fruit jam or puree. Typical flavorings include strawberry, raspberry, blueberry, apricot, pomegranate, peach, pineapple, lemon, orange, and apple. Generally, fruit flavorings include any combination of fruit extract, fruit preserve or puree, and sweetener, starch, stabilizer, natural and / or artificial flavor, color, preservative, water, and citric acid or other suitable acid for pH control.

[0172] For oral preparations, the compositions can be used alone or in combination with appropriate additives to prepare tablets, powders, granules or capsules, e.g., with conventional additives, such as lactose, mannitol, corn starch or potato starch; with binders, such as crystalline cellulose, cellulose derivative, acacia, corn starch or gelatins; with disintegrators, such as corn starch, potato starch or sodium carboxymethyl cellulose; with lubricants, such as talc or magnesium stearate; and if desired, with diluents, buffering, moistening or flavoring agents.

[0173] The compositions can be formulated into a liquid preparation by dissolving, suspending or emulsifying the compositions in an aqueous or non-aqueous solvent, such as vegetable or other similar oils, synthetic aliphatic, aldehyde, or ester solvents, or glycerol, and if desired, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives. The compositions can be used in the form of aerosol preparations, e.g., for inhalation. The compositions can be made into suppositories by mixing with a variety of excipients, such as emulsifiers or wetting agents, e.g., as mono- or di-glycerides, or stearic acid salts.

[0174] Unit dosage forms for oral administration can be provided, such as syrups, elixirs, and suspensions, each containing a predetermined amount of the composition of the present application. Similarly, unit dosage forms for injection or intravenous administration can include a composition containing a compound of the present application in a solution, in which each dosage unit, for example, mL or L, contains a predetermined amount of the composition of the present application.

[0175] Table 1: Representative powder composition I:

[0176] Composition per 100 g Protein 10-80g Carbohydrate 20-40g Fat 0-20g Polyunsaturated fatty acids 0-5g Fiber 0-5g Vitamins 0-100% of the respective RDA Minerals 0-100% of the respective RDA Folic acid 0-1000 μg Nicotinic acid 0-100 mg Creatine 0-20g Ketones (e.g. beta-hydroxybutyric acid) 0.025-5g Urolithin A 0.025-5g

[0177] Table 2: Representative powder composition II:

[0178] Composition per 20 g Vitamins 0-100% of the respective RDA Minerals 0-100% of the respective RDA Folic acid 0-1000 μg Nicotinic acid 0-100 mg Creatine 0-2g Excipient filler 0-4.5g Ketones (e.g. beta-hydroxybutyric acid) 0.025-5g Urolithin A 0.025-5g

[0179] Bulk powders are usually supplied with instructions telling the subject how much powder to take in one dose. For example, a bulk powder can be supplied in a container with a scoop of the required size attached so that the correct amount of powder can be measured out. The powder can be taken directly, mixed with food, or added to water or fruit juice to make a drink.

[0180] Table 3: Representative beverage composition I:

[0181]

[0182] Table 4: Representative beverage composition II:

[0183] Composition per 30 mL Vitamins 0-100% of the respective RDA Minerals 0-100% of the respective RDA Folic acid 0-500 μg Nicotinic acid 0-20 mg Ketones (e.g. beta-hydroxybutyric acid) 0.025-5g Urolithin A 10-5000 mg

[0184] Table 5: Representative Stick Compositions I:

[0185]

[0186]

[0187] Table 6: Representative Stick Compositions II:

[0188] Composition per 35 g bar Vitamins 0-100% of the respective RDA Minerals 0-100% of the respective RDA Nicotinic acid 0-100 mg L-carnitine 0-500 mg Ketones (e.g. beta-hydroxybutyric acid) 0.025-5g Urolithin A 10-5000 mg

[0189] Table 7: Representative Yogurt Compositions

[0190]

[0191] Representative Soft Capsule Formulations (i.e., soft capsules containing a gelatin shell and a fill containing Urolithin A)

[0192] Table 8: Representative Fills for Soft Capsules

[0193]

[0194]

[0195] Table 9: Representative Shells for Soft Capsules

[0196]

[0197] The compositions of the present application can be administered as a single treatment, or more commonly, as a series of treatments. In one example, a subject takes a dose before or after exercise. For subjects who are unable to exercise, a dose of the composition can be taken once, twice, or three times a day, or once, twice, three, four, five, or six times a week, for example. It will also be appreciated that the effective dose of the compound can be increased or decreased over the course of a particular treatment.

[0198] Examples

[0199] The following examples illustrate the present application.

[0200] Example 1: Effect of the combination of ketone bodies 3-β-hydroxybutyric acid and acetoacetate with urolithin A on cellular energy production

[0201] 3-β-hydroxybutyrate and acetoacetate are ketone bodies produced by the liver during ketogenesis and can be utilized by peripheral tissues to produce ATP. To achieve this, 3-β-hydroxybutyrate and acetoacetate are imported into mitochondria and converted to acetyl-CoA. Acetyl-CoA can then enter the tricarboxylic acid cycle and be converted to citrate by citrate synthase. NAD + and FADH co-factors are then reduced, which will allow ATP production through oxidative phosphorylation.

[0202] C2C12 cells were incubated with 20 mM Urolithin A or vehicle (with or without 3-β- hydroxybutyrate, acetoacetate or a combination of both) for 24 hours to assess the effect of 3-β-hydroxybutyrate and acetoacetate alone or in combination with UA on mitochondrial function.

[0203] ATP and citrate synthase

[0204] Total ATP content and citrate synthase (CS) enzyme activity were measured as described in Mouchiroud, L. et al. Cell 154, 430-441 (2013). Briefly, total ATP content was measured by Luminescent cell viability assay (Promega) was measured and CS using CS assay kit (Sigma).

[0205] Respiratory capacity

[0206] Cells were incubated with test compounds under high glucose media (regular cell culture conditions) and low glucose conditions using sequential injections of oligomycin, FCCP (carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone) and rotenone / antimycin. This determines maximum respiratory capacity and uncoupled respiration.

[0207] To distinguish between anaerobic glycolytic ATP and oxidative phosphorylation ATP, Seahorse XF ATP protocol was also employed (https: / / www.agilent.com / en / products / cell-analysis / seahorse-xf-atp).

[0208] Fatty acid oxidation

[0209] Measurements were performed following standard protocol for fatty acid oxidation (https: / / www.agilent.com / cs / pubimages / misc / XF_FAO_Assay_Technical_Brief.pdf). Briefly, once C2C12 myotubes were fully differentiated in 96-well plates, they were incubated in 0.5 mM glucose, 1 mM GlutaMAX TM, 0.5mM carnitine based on the substrate restriction culture medium of DMEM, process 24 hours 45 minutes.On the mensuration day, cell is measured culture medium (111mM NaCl, 4.7mM KCl, 1.25mM CaCl , 2mM MgSO , 1.2mM NaH PO , 2.5mM glucose, 0.5mM carnitine and 5mM HEPES, at 37 ℃, be adjusted to pH 7.4) washed twice.Then cell is put into non-CO incubator and incubated 30 minutes.Before starting to analyze, with the ultimate density of 166 μ M palmitates, cell is supplemented with XF palmitate-BSA FAO substrate (seahorse biological science company (Seahorse Bioscience)).Oxygen consumption rate is measured at basal level and after adding FCCP.

[0210] Example 2: Effects of the combination of ketone bodies 3-β-hydroxybutyrate and acetoacetate with urolithin A on the expression of marker genes for mitochondrial function

[0211] C2C12 cells were incubated with 20 μM urolithin A or vehicle (with or without 3-β-hydroxybutyrate, acetoacetate, or a combination of both) 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.

[0212] Measuring gene expression by RNA-seq

[0213] Total RNA was extracted from C2C12 cells using TRIzol (Thermo Fisher Scientific) and expressed using The micro kit (QIAGEN, 74104) was purified by column. The total RNA sample concentration, RIN, 28S / 18S and size were detected using an Agilent Bioanalyzer 2100 (Agilent RNA 6000Nano kit). The purity of the sample was tested by NanoDropTM. RNA-seq library preparation was performed as described in D'Amico et al., Mol Cell 2019. RNA-seq runs were performed using an Illumina HiSeq4000 sequencing platform with a single read (1×50bp) and 30 million reads (+ / -3%). The obtained reads were first trimmed to remove adapter sequences and then mapped to the human genome (GRCh37). Differential gene expression (DGE) was calculated and genes with a nominal p-value <0.05 were selected, and gene set enrichment analysis was run using the GSEA tool. The gene set was taken from the MSIGDB C5 GO collection. Gene sets related to mitochondrial biology were selected with a false discovery rate (FDR) < 0.25.

[0214] Example 3: Synergistic anti-inflammatory effect of Urolithin A with betaDL-beta-hydroxybutyric acid sodium salt (BHB)

[0215] Methods

[0216] C2C12 myoblast cells (reference CRL-1772) purchased from the American Type Culture Collection (ATCC) were seeded in 96-well plates (Greiner, Kremsmünster, Austria) at 8000 cells per well using DMEM Glutamax 4.5g D-glucose (Gibco, reference 31966021) supplemented with 10% fetal bovine serum (FBS) (PanBiotech, reference 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 31966021), 2% horse serum (Thermo Fisher, reference 160501229), 1% PenStrep and 1% HEPES). All media were sterilized with a 0.45 pm filtration unit (VWR). After 4 days of differentiation, cells were treated for 24 hours with the indicated doses of: 1) Urolithin A (UA), DL-beta-hydroxybutyric acid sodium salt (BHB) (Sigma, reference: H6501-5GA) or a combination of both suspended in DMSO. On day 5, in addition to the previous treatment, a cytokine mix (“cytomix”) was added for 24 hours using 5 ng of each cytokine (mIFN-g (Roche, reference 11276905001), TNF-a (Peprotech, reference 300-01A) and IL-b (Proteintech, reference 200-01B) respectively). Media were collected and ELISA for interleukin-6 was performed according to the instructions provided by the supplier (Proteintech, reference KE10007). Finally, absorbance at 450 nm wavelength was measured using a microplate reader (Fluostar Optima, BMG Labtech).

[0217] Results

[0218] C2C12 cells exposed to a pro-inflammatory cytokine cocktail (cytomix) resulted in a significant increase in interleukin 6 (IL-6) secretion, measured as IL-6 concentration in cell culture media. Urolithin A pre-treatment dose-dependently reduced IL-6 secretion, with a 25% dose-dependent reduction at low dose 6.25 uM and 60% reduction at high dose 12.5 uM Figure 1

[0219] To explore the potential synergistic effect of Urolithin A (UA) and beta-hydroxybutyrate (BHB), cells were treated with these two compounds alone or in combination, using a fixed sub-functional UA dose of 6.25 uM and 5uM or 25uM BHB.

[0220] Low dose BHB alone (5 uM) had no effect on IL-6 secretion (-0.007%). Surprisingly, the combination of low dose UA (6.25 uM) and low dose BHB (5 uM) significantly reduced the inflammatory marker by 56%. This was also statistically significant compared to the 25% reduction observed with UA 6.25 uM alone (Table 1).

[0221] Higher dose of BHB (2 uM) significantly reduced IL-6 secretion by 32%. The combination of low dose UA (6.25 uM) and high dose BHB reduced IL-6 secretion to 58%. This suggests that a fixed dose of UA is able to produce a synergistic effect with BHB within a certain dose range Figure 2 and Table 1).

[0222] Ketones 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 of 2 mM (PMID: 25686106) or 250 uM (PMC6402511) and above. The combination of Urolithin A 6.25 uM and BHB 5 uM results suggest that UA is able to unlock the potential of otherwise ineffective low BHB doses within a specific and surprising dose window.

[0223] Table 1.

[0224]

[0225] BHB - beta-hydroxybutyrate

[0226] CTX - cytomix

[0227] UA - Urolithin A​

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 combination according to claim 1 or claim 2 or the composition according to claim 3, wherein the urolithin is a compound of formula (I) wherein A, B, C, D, W, X, Y and Z are each independently selected from H and OH (or a salt, prodrug, metabolite or derivative thereof).

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

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

7. A combination or composition according to 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. The combination or composition according to 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 body is acetoacetic acid and / or β-hydroxybutyric acid and / or acetone.

10. The combination or composition according to any one of claims 1 to 9, wherein the ketone body is β-hydroxybutyrate (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 beta-hydroxybutyrate.

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

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

15. Use of a combination or composition according to any one of claims 1 to 11 for 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. 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 muscle function decline, increasing muscle mass and / or reducing muscle wasting.

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

19. A combination or composition according to any one of claims 1 to 11 for use in enhancing 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; and (ii) A separate composition comprising a urolithin or a salt, prodrug, metabolite or derivative thereof.

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

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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