Compositions Comprising Urolithins for Treating Muscle Wasting and Renal Dysfunction

Abstract

The present disclosure generally relates to methods and compositions comprising a urolithin for treating or preventing diseases or conditions associated with muscle wasting and / or impaired kidney function. Additionally, methods and compositions comprising a urolithin for increasing urinary creatinine and / or decreasing the albumin / creatinine ratio and / or increasing albumin reabsorption. Additionally, methods and compositions comprising a urolithin for improving muscle lean mass and / or improving muscle fiber size and / or improving the levels of at least one amino acid are disclosed.

Description

[Technical field]

[0001] The present disclosure generally relates to methods and compositions comprising urolithins for treating or preventing diseases or conditions associated with muscle wasting and / or renal dysfunction. Additionally disclosed are methods and compositions comprising urolithins for increasing urinary creatinine, and / or decreasing albumin / creatinine ratio, and / or increasing albumin reabsorption. Additionally disclosed are methods and compositions comprising urolithins for improving muscle lean mass, and / or improving muscle fiber size, and / or improving the level of at least one amino acid. Also disclosed are methods and compositions comprising urolithins for improving the level of at least one metabolic product, and / or improving the level of at least one nucleotide, and / or improving the level of at least one nicotinamide adenine dinucleotide, and / or increasing the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increasing the level of succinate and / or malate, and / or increasing the level of phosphocreatine. [Background technology]

[0002] The loss of muscle mass that occurs during muscle wasting can be characterized by catabolic muscle protein breakdown, which, whether caused by high rates of protein degradation or low rates of protein synthesis, leads to loss of muscle mass and muscle wasting.

[0003] Muscle wasting due to muscle wasting is associated with chronic, neurological, genetic or infectious pathologies, disorders, diseases or conditions, particularly in chronic kidney disease (CKD), end-stage renal disease (ESRD), metabolic dysfunction-induced muscle wasting, muscle weakness and / or renal failure due to dialysis, diabetes, intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, protein-energy wasting, among others.

[0004] Muscle wasting, characterized by a decline in muscle mass, strength and function, is a common complication of CKD and significantly increases the risk of morbidity and mortality in this population. Many complications associated with reduced renal function and lifestyle factors activate catabolic pathways and reduce muscle regeneration, resulting in substantial protein wasting.

[0005] In CKD, there is a gradual and progressive decline in the kidney's ability to excrete waste products, concentrate urine, reabsorb proteins and amino acids, and conserve electrolytes. Unlike acute renal failure, where renal function is sudden but reversible, in chronic kidney disease renal function irreversibly progresses and deteriorates toward end-stage renal disease (ESRD). CKD results from heterogeneous disease pathways that irreversibly alter kidney function and structure over months or years.

[0006] Diabetes and hypertension are the main causes of CKD in all high- and middle-income countries, as well as in many low-income countries. The incidence, prevalence, and progression of CKD are likely epigenetic influenced, and vary within countries, by ethnic group, and by social determinants of health. Many people are asymptomatic or have nonspecific symptoms such as lethargy, itching, or loss of appetite. Diagnosis is generally made after incidental detection from screening tests such as urine dipsticks or blood tests, or when symptoms become severe. The best available indicator of general kidney function is the glomerular filtration rate (GFR). Disease and management are classified according to stages of disease severity, assessed by GFR and albuminuria, and clinical diagnosis (cause and pathology). The presence of proteinuria is associated with an increased risk of progression of CKD. Diagnosis of CKD is based on the chronic decline in kidney function and confirmation of structural kidney damage.

[0007] CKD is diagnosed using a staging system that documents the amount of available kidney function (stage 1 = normal kidney function) and patients often have no symptoms in the early stages. Stage 5 of CKD is ESRD, where the kidneys are in complete or near complete failure, and usually occurs when kidney function is less than 10% of baseline.

[0008] [Summary of the invention] In a first aspect, the disclosure provides a method of treating and / or preventing a disease or condition associated with muscle wasting and / or kidney dysfunction. The method comprises administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, muscle wasting and / or kidney dysfunction is treated and / or prevented at early stages of chronic kidney disease and / or at later stages of chronic kidney disease.

[0009] In a second aspect, the disclosure provides a method for improving lean muscle mass in a subject in need thereof comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, lean muscle mass is improved in early stages of chronic renal disease and / or in later stages of chronic renal disease.

[0010] In some embodiments of the foregoing aspects, the muscle is the tibialis anterior and / or the quadriceps.

[0011] In a third aspect, the disclosure provides a method for increasing albumin reabsorption in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, albumin reabsorption is increased in early stages of chronic kidney disease and / or in later stages of chronic kidney disease.

[0012] In a fourth aspect, the disclosure provides a method for increasing urinary creatinine in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, urinary creatinine is increased in early stages of chronic kidney disease and / or in late stages of chronic kidney disease.

[0013] In a fifth aspect, the disclosure provides a method for reducing the albumin / creatinine ratio in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, the albumin / creatinine ratio is reduced in early stages of chronic kidney disease and / or in late stages of chronic kidney disease.

[0014] In a sixth aspect, the disclosure provides a method for increasing urinary carnosine in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, urinary carnosine is increased in early chronic kidney disease and / or in late chronic kidney disease.

[0015] In a seventh aspect, the disclosure provides a method for increasing urinary anserine in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, urinary anserine is increased in early chronic kidney disease and / or in late chronic kidney disease.

[0016] In an eighth aspect, the disclosure provides a method for increasing urinary S-adenosylmethionine in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, urinary S-adenosylmethionine is increased in early chronic kidney disease and / or late chronic kidney disease.

[0017] In a ninth aspect, the disclosure provides a method for improving femoral bone mass in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In one embodiment, femoral bone mass is improved in early stages of chronic renal disease and / or in late stages of chronic renal disease.

[0018] In some embodiments of the foregoing aspects, the muscle is the tibialis anterior and / or the quadriceps.

[0019] In a tenth aspect, the disclosure provides a method for improving muscle fiber size in a subject in need thereof comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

[0020] In an eleventh aspect, the disclosure provides a method for improving the level of at least one amino acid in muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving the level of at least one amino acid in muscle of a subject in need thereof.

[0021] In a twelfth aspect, the disclosure provides a method for improving the level of at least one metabolic product in muscle of a subject in need thereof, comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

[0022] In a thirteenth aspect, the disclosure provides a method for improving levels of at least one nucleotide in muscle of a subject in need thereof, comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

[0023] In a fourteenth aspect, the disclosure provides a method for improving levels of at least one nicotinamide adenine dinucleotide in muscle of a subject in need thereof, comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

[0024] In a fifteenth aspect, the present disclosure provides a method for increasing glutathione (GSH) and / or glutathione disulfide (GSSG) levels in muscle of a subject in need thereof, comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

[0025] In a sixteenth aspect, the disclosure provides a method for increasing succinate and / or maleate levels in muscle of a subject in need thereof, comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

[0026] In a seventeenth aspect, the disclosure provides a method for increasing the level of phosphocreatine in muscle of a subject in need thereof, comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

[0027] In some embodiments of the foregoing aspects, the subject has a disease or condition selected from chronic kidney disease, metabolic induced muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

[0028] In other embodiments, the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

[0029] In further embodiments, the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

[0030] In some embodiments, the urolithin is micronized.

[0031] In other embodiments, the urolithin is urolithin A.

[0032] In an eighteenth aspect, the present disclosure provides a nutritional composition for use in the treatment and / or prevention of a disease or condition associated with muscle wasting and / or kidney dysfunction. The nutritional composition comprises a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof. Preferably, the nutritional composition contains an amount of urolithin effective to treat and / or prevent a disease or condition associated with muscle wasting and / or kidney dysfunction in a subject in need thereof.

[0033] In one embodiment, the composition is selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low volume liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

[0034] In another embodiment, the composition is formulated for oral administration.

[0035] In a nineteenth aspect, the disclosure provides a unit dosage form for use in a method for treating and / or preventing a disease or condition associated with muscle wasting and / or impaired kidney function. The unit dosage form comprises a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. Preferably, the unit dosage form contains an amount of urolithin effective to treat and / or prevent a disease or condition associated with muscle wasting and / or impaired kidney function in a subject in need thereof.

[0036] In one embodiment, the unit dosage form is selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low volume liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

[0037] In another embodiment, the unit dosage form is formulated for enteral administration.

[0038] In some embodiments of the eighteenth and nineteenth aspects, the composition and / or unit dosage form contains an amount of urolithin effective to increase urinary creatinine, and / or decrease the albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve muscle lean mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

[0039] In other embodiments of the eighteenth and nineteenth aspects, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to improve lean mass of muscle and / or improve muscle fiber size and / or improve levels of at least one amino acid.

[0040] In further embodiments of the eighteenth and nineteenth aspects, the composition comprising a urolithin or the amount of a urolithin or a pharmacologically acceptable salt thereof is effective to improve the level of at least one metabolic product, and / or improve the level of at least one nucleotide, and / or improve the level of at least one nicotinamide adenine dinucleotide, and / or increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increase the level of succinate and / or malate, and / or increase the level of phosphocreatine.

[0041] An advantage of one or more aspects and embodiments provided by the present disclosure is that it aids in maintaining healthy muscle mass.

[0042] Another advantage of one or more aspects and embodiments provided by the present disclosure is that it helps stabilize renal dysfunction.

[0043] Yet another advantage of one or more aspects and embodiments provided by the present disclosure is that it aids in restoring kidney function.

[0044] An advantage of one or more aspects and embodiments provided by the present disclosure is that it helps improve lean muscle mass.

[0045] Another advantage of one or more aspects and embodiments provided by the present disclosure is that they help increase albumin reabsorption.

[0046] Yet another advantage of one or more aspects and embodiments provided by the present disclosure is that it aids in increasing urinary creatinine.

[0047] An advantage of one or more aspects and embodiments provided by the present disclosure is that they aid in reducing the albumin / creatinine ratio.

[0048] Another advantage of one or more aspects and embodiments provided by the present disclosure is that they help increase urinary carnosine.

[0049] Yet another advantage of one or more aspects and embodiments provided by the present disclosure is that it helps increase urinary anserine.

[0050] An advantage of one or more aspects and embodiments provided by the present disclosure is that they help increase urinary S-adenosylmethionine.

[0051] Another advantage of one or more aspects and embodiments provided by the present disclosure is that they aid in improving femoral mass.

[0052] An advantage of one or more aspects and embodiments provided by the present disclosure is that it improves muscle endurance and / or efficiency.

[0053] Another advantage of one or more aspects and embodiments provided by the present disclosure is that it improves muscle fiber size.

[0054] Yet another advantage of one or more aspects and embodiments provided by the present disclosure is improving amino acid levels in muscle.

[0055] An advantage of one or more aspects and embodiments provided by the present disclosure is that it reduces muscle atrophy.

[0056] Another advantage of one or more aspects and embodiments provided by the present disclosure is that it increases nucleotides for muscle bioenergetics and nucleotides important for cell signaling.

[0057] Additional features and advantages are described herein, and will be apparent from the drawings and detailed description that follow. [Brief description of the drawings]

[0058] [Figure 1a] Albumin / creatine ratio measured in mice after 5 weeks (midpoint) of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day, compared to vehicle (diabetic, db / db+V group) and control (control, db / m+V group) after 5 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Creatinine was measured using a Cobas Integra autoanalyzer. Urinary albumin excretion was measured by ELISA kit. [Figure 1b] Albumin / creatine ratio measured in mice after 10 weeks (endpoint) of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle group (diabetic, db / db+V group) and control group (control, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Creatinine was measured using a Cobas Integra autoanalyzer. Urinary albumin excretion was measured by ELISA kit. [Diagram 2]Urinary creatinine measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) is shown compared to the vehicle group (diabetic, db / db+V group) and the control group (control, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Urinary creatinine was measured using a Cobas Integra automated analyzer. [Diagram 3] The progression of albuminuria measured in mice at baseline (time zero), after 5 weeks (midpoint) and after 10 weeks (endpoint) of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to the vehicle group (diabetic, db / db+V group) and the control group (control, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Urinary albumin excretion was measured by ELISA kit. [Figure 4] Figure 1 shows muscle lean mass (g) measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (control, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Muscle lean mass was measured using EchoMRI. [Figure 5a] The muscle mass (mg) of the tibialis anterior muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) is shown compared to the vehicle group (diabetic, db / db+V group) and the control group (control, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. The muscle mass of the tibialis anterior muscle was measured by weighing on a precision balance. [Figure 5b]Quadriceps muscle mass (mg) measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (control, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Quadriceps muscle mass was measured by weighing on a precision balance. [Figure 6]Urinary carnosine measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day is shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Urinary carnosine was measured by a semi-quantitative method using cold methanol:water:chloroform (5:3:5 (v / v)) extraction to separate polar and non-polar metabolites. Furthermore, a protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4° C. and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH) in H2O (pH ∼9.3), and solvent B was acetonitrile (ACN). Elution of metabolites, including carnosine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on-the-fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 7]Urinary anserine measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day is shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Urinary anserine was measured by a semi-quantitative method using cold methanol:water:chloroform (5:3:5 (v / v)) extraction to separate polar and non-polar metabolites. Furthermore, a protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4° C. and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH) in H2O (pH ∼9.3), and solvent B was acetonitrile (ACN). Elution of metabolites, including anserine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 8]Urinary S-adenosylmethionine measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day is shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Urinary S-adenosylmethionine was measured by a semi-quantitative method using cold methanol:water:chloroform (5:3:5 (v / v)) extraction to separate polar and non-polar metabolites. Furthermore, a protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4° C. and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including S-adenosylmethionine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 9]Femur mass measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) is shown compared to vehicle (diabetic, db / db+V group) and control (control, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Femur mass was measured by weighing on a precision balance. [Figure 10] The size distribution of fibers measured within the tibia in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) is shown compared to vehicle (diabetic, db / db+V group) and control (control, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Fiber size distribution was assessed after tibialis anterior muscle cryosections and stained for laminin protein and myonuclei. All slides were acquired on an Olympus VS120 slide scanner microscope. Myofiber size was calculated in Min Feret using an automated image processing algorithm developed in-house using QuPath software and the Fiji tool open-CSAM. [Figure 11a]Isoleucine levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Isoleucine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of isoleucine-containing metabolites was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 11b]Methionine levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Methionine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of methionine-containing metabolites was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 11c]Lysine levels measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Lysine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). The elution of lysine-containing metabolites was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 11d]Tyrosine levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Tyrosine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of tyrosine-containing metabolites was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 11e]Proline levels measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Proline levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of proline-containing metabolites was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 11f]Alanine levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Alanine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of alanine-containing metabolites was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 11g]Glycine levels measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Fiber size distribution was measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of glycine-containing metabolites was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 12a]N-acetyl-DL-serine levels measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. N-acetyl-DL-serine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including N-acetyl-DL-serine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 12b]N-acetyl-DL-arginine levels measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. N-acetyl-L-arginine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including N-acetyl-L-arginine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 12c]N-acetylglutamic acid levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. N-acetylglutamic acid levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including N-acetylglutamic acid, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 13a]N,N-dimethylglycine levels measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. N,N-dimethylglycine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including N,N-dimethylglycine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 13b]S-adenosylmethionine (SAM) levels measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. NS-adenosylmethionine (SAM) levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including S-adenosylmethionine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 14a]2-Hydroxybutyrate levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) groups after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. 2-Hydroxybutyrate levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including 2-hydroxybutyrate, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 14b]The trans-urocanic acid levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to the vehicle group (diabetic, db / db+V group) and the control group (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. The trans-urocanic acid levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including trans-urocanic acid, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 15a]Anserine levels in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Anserine levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites including anserine in muscle was analyzed with an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 15b]Figure 1 shows muscle carnosine levels measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Muscle carnosine levels were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including carnosine levels in muscle, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 16a]Adenosine triphosphate (ATP) levels in muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Adenosine triphosphate (ATP) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). The elution of metabolites containing adenosine triphosphate was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 16b]Adenosine diphosphate (ADP) levels in muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Adenosine diphosphate (ADP) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of adenosine diphosphate-containing metabolites in muscle was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 16c]Guanosine triphosphate (GTP) levels in muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Guanosine triphosphate (GTP) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of guanosine triphosphate-containing metabolites in muscle was analyzed with an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 16d]Guanosine diphosphate (GDP) levels in muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Guanosine diphosphate (GDP) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites including guanosine diphosphate in muscle was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 16e]Uridine triphosphate (UTP) levels in muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Uridine triphosphate (UTP) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites including uridine triphosphate in muscle was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Fig. 16f]Cytidine triphosphate (CTP) levels in muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Cytidine triphosphate (CTP) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites containing cytidine triphosphate in muscle was analyzed with an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 16g]Flavin adenine dinucleotide (FAD) levels in muscle measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Flavin adenine dinucleotide (FAD) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites including flavin adenine dinucleotide in muscle was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 17a]NAD+ levels in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. NAD+ levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites including NAD+ in muscle was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 17b]NADP+ levels in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. NADP+ levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites including NADP+ was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 18a]Glutathione (GSH) in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Glutathione (GSH) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including GSH, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 18b]Glutathione disulfide (GSSG) in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Glutathione disulfide (GSSG) levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites including GSSG was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV) alternated on the fly were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 19a]Succinate in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day is shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Succinate levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including succinates, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 19b]Maleate levels in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Maleate levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including maleate, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. [Figure 20]Phosphocreatine levels in muscle measured in mice after 10 weeks of treatment with Urolithin A (+UA, db / db+UA group) at a dose of 50 mg / kg / day are shown compared to vehicle (diabetic, db / db+V group) and control (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose. Phosphocreatine levels in muscle were measured using liquid-liquid extraction with 13C-yeast as an internal standard. Muscle extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min. Furthermore, the protein layer remained in the middle between the two phases. The polar phase was dried overnight in a vacuum centrifuge at 4°C and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations. 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was H2O (pH ~ 9.3) containing 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH), and solvent B was acetonitrile (ACN). Elution of metabolites, including phosphocreatine, was analyzed on an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. Negative ion mode (3 kV) and positive ion mode (3.5 kV), alternated on the fly, were used for ionization. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control, data acquisition, and processing. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0059] definition Some definitions are provided below. However, definitions may be found in the "Embodiments" section below, and the heading "Definitions" above does not imply that such disclosure in the "Embodiments" section is not a definition.

[0060] All percentages stated herein are by total weight of the composition unless otherwise indicated. As used herein, "about," "approximately," and "substantially" are understood to refer to numbers within a certain range of numerical values, for example, within the range of -10% to +10% of the referenced number, preferably within the range of -5% to +5% of the referenced number, more preferably within the range of -1% to +1% of the referenced number, and most preferably within the range of -0.1% to +0.1% of the referenced number. All numerical ranges herein should be understood to include all integers or fractions within the range. Furthermore, these numerical ranges should be interpreted as supporting claims directed to any number or subset of numbers within the range. For example, a disclosure of 1 to 10 should be interpreted as corresponding to ranges of 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, 3.5 to 9.9, etc.

[0061] As used in this disclosure and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context dictates otherwise. Thus, for example, reference to "a component" or "the component" includes two or more components.

[0062] The terms "comprise", "comprises", and "comprising" should be construed as inclusive rather than exclusive. Similarly, the terms "include", "including", and "or" should all be construed as inclusive unless such interpretation is clearly prevented by the context. However, the compositions disclosed herein may not include elements not specifically disclosed herein. Thus, disclosure of an embodiment using the term "comprising" includes disclosure of embodiments "consisting essentially of" and "consisting of" the specified components. A composition or unit dosage form "consisting essentially of" means that the referenced component comprises at least 50% by weight, preferably at least 75% by weight, more preferably at least 85% by weight, and most preferably at least 95% by weight of the referenced component.

[0063] The term "and / or" used in the context of "X and / or Y" should be interpreted as "X" or "Y" or "X and Y". Similarly, "at least one of X or Y" should be interpreted as "X" or "Y" or "X and Y". For example, "muscle attenuation and / or renal dysfunction" should be interpreted as "muscle attenuation" or "renal dysfunction" or "both muscle attenuation and renal dysfunction".

[0064] As used herein, the terms "examples" and "such as," especially when followed by a list of terms, are merely exemplary and illustrative and should not be considered as exclusive or inclusive. As used herein, a condition "associated with" or "linked with" another condition means that the conditions occur simultaneously, preferably that the conditions are caused by the same underlying condition, and most preferably that one of the specified conditions is caused by the other specified condition.

[0065] "Prevention" includes reducing the risk and / or severity of a condition or disease. The terms "treatment", "treat" and "to alleviate" include both prophylactic or preventive treatments (which prevent and / or delay the onset of the targeted condition or disorder) and curative, therapeutic or disease-modifying treatments, including therapeutic measures that cure, delay, attenuate the symptoms and / or halt the progression of a diagnosed condition or disease; as well as treatment of patients who are ill or diagnosed as suffering from a disease or medical condition, in addition to treatment of patients at risk of or suspected of having a disease. The terms do not necessarily imply that a subject is treated until cured. The terms "treatment" and "treat" also refer to maintaining and / or promoting the health of individuals who are not afflicted with a disease but who may be susceptible to an unhealthy condition. The terms "treatment / therapy", "treat / treat" and "alleviating" are also intended to include the synergistic or otherwise potentiating action of one or more primary preventative or therapeutic measures. The terms "treatment / therapy", "treat / treat" and "alleviating" are further intended to include the dietary management of a disease or condition, or dietary management for the prophylaxis or prevention of a disease or condition. Treatment may be patient-related or physician-related.

[0066] A "subject" or "individual" is a mammal, preferably a human. The term mammal may include, but is not limited to, pets or livestock. In particular, the term "livestock" may include, but is not limited to, horses (e.g., pets or horses undergoing medical treatment), or cattle or poultry (e.g., cattle or poultry used in agriculture). In particular, the term "pet" may include, but is not limited to, senior pets (e.g., cats over 10 years old or dogs over 7 years old), obese pets, diabetic pets, and CKD pets.

[0067] As used herein, an "effective amount" is an amount that prevents a deficiency, treats a disease or medical condition in an individual, or, more generally, alleviates symptoms, manages the progression of a disease, or provides a nutritional, physiological, or medical benefit to an individual. The relative terms "improved," "increased," "enhanced," and the like refer to the effect of the compositions disclosed herein, i.e., compositions comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof.

[0068] The terms "food," "food product," and "food composition" refer to a product or composition intended for consumption by an individual, such as a human, and providing at least one nutrient to such an individual. Food products typically contain at least one of protein, lipid, carbohydrate, and optionally one or more vitamins and minerals. The compositions of the present disclosure, including the many embodiments described herein, may comprise, consist of, or essentially comprise the elements disclosed herein, as well as any additional or optional ingredients, components, or elements described or not described herein that are useful in the diet.

[0069] As used herein, the term "unit dosage form" refers to physically discrete units suitable as dosage units for human and animal subjects, each unit containing a predetermined amount of a composition disclosed herein, in an amount sufficient to produce the desired effect, together with a pharma- ceutically acceptable diluent, carrier, or vehicle. The specifications for the unit dosage forms depend on the particular compound and / or composition used, the effect to be achieved, and the pharmacodynamics associated with each compound and / or composition in the host.

[0070] "Chronic kidney disease (CKD)" refers to renal impairment (i.e., albuminuria) or reduced renal function (i.e., glomerular filtration rate <60 mL / min / 1.73 m), regardless of clinical diagnosis. 2CKD is classified into five stages based on GFR: >90 mL / min / 1.73 m 2 Ultra (Stage 1), 60-89 mL / min / 1.73 m 2 (Stage 2), 30-59 mL / min / 1.73 m 2 (Stage 3), 15–29 mL / min / 1.73 m 2 (Stage 4), and 15 mL / min / 1.73 m 2 Less than (stage 5).

[0071] "Early stage of chronic kidney disease" is 60-89mL / min / 1.73m 2 (Stage 2), 30-59 mL / min / 1.73 m 2 (Stage 3) includes stage 2 and stage 3 chronic kidney disease based on the above. Preferably, "early stage chronic kidney disease" includes a blood glucose level of 60 to 89 mL / min / 1.73 m 2 This includes stage 2 chronic kidney disease based on

[0072] "Late stage chronic kidney disease" is 15-29mL / min / 1.73m 2 (Stage 4), and 15 mL / min / 1.73 m 2 (Stage 5) includes stage 4 and stage 5 chronic kidney disease based on a blood glucose level of 15-29 mL / min / 1.73 m 2 This includes stage 4 chronic kidney disease based on

[0073] "End stage renal disease (ESRD)" includes the condition of an individual with CKD who requires renal replacement therapy. Preferably, ESRD is defined as a condition in which the GFR is below 15 mL / min / 1.73 m 2 Includes stage 5 chronic kidney disease based on less than

[0074] "Metabolic muscle wasting" involves a prolonged catabolic state in which protein breakdown in muscle exceeds the rate of protein synthesis.

[0075] "Diabetes" includes both types I and II of the disease. Non-limiting examples of risk factors for diabetes include a waistline greater than 40 inches for men or greater than 35 inches for women, blood pressure equal to or greater than 130 / 85 mmHg, triglycerides greater than 150 mg / dL, fasting blood glucose greater than 100 mg / dL, or high density lipoprotein less than 40 mg / dL for men or less than 50 mg / dL for women.

[0076] "Pompe disease" includes classical infantile, non-classical infantile, and late-onset Pompe disease. In children with classical or non-classical infantile-onset Pompe disease, acid α-glucosidase activity is generally less than about 1% of normal. In individuals with late-onset Pompe disease, acid α-glucosidase activity is generally less than about 40% of normal.

[0077] "Metabolic acidosis" encompasses a decrease in serum pH and an abnormal serum bicarbonate concentration of <22 mEq / L, below the normal range of 22 to 29 mEq / L.

[0078] "Methylmalonic aciduria" refers to an inherited disorder in which the body is unable to properly digest certain fats and proteins, as well as the amino acids methionine, threonine, isoleucine and valine, thus resulting in the buildup of toxic amounts of methylmalonic acid in the blood.

[0079] "Disuse atrophy" refers to a temporary condition in which unused muscles are properly exercised after a cast is removed from a limb, or when a person regains enough strength to move after being bedridden for a period of time.

[0080] "Protein-energy wasting" refers to the reduction in body protein mass and fuel reserves in a subject due to maladaptive metabolic conditions, including non-specific inflammatory processes, transient intercurrent catabolic disorders, nutrient loss into the dialysate, acidemia, insulin resistance, endocrine disorders such as growth hormone and insulin-like growth factor-1, hyperglucagonemia, hyperparathyroidism, and blood loss into the hemodialyzer, into the feces, or through blood sampling.

[0081] "Metabolic product" or "metabolite" refers to a metabolic intermediate or end product. The term "metabolic product" or "metabolite" may relate to a metabolic intermediate or end product related to muscle metabolism and / or growth. In particular, the term "metabolic product" or "metabolite" may relate to a metabolic intermediate or end product in muscle.

[0082] Embodiment In a first aspect, the disclosure provides a method of treating and / or preventing a disease or condition associated with muscle wasting and / or impaired kidney function. The method comprises administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method of treating and / or preventing a disease or condition associated with muscle wasting and / or impaired kidney function.

[0083] In one embodiment, the method is a method of treating at least one disease or condition associated with muscle wasting and / or impaired renal function, the method comprising administering a composition comprising a urolithin, or a composition comprising a prophylactically effective amount of a urolithin or a pharmacologically acceptable salt thereof, to a subject having at least one disease or condition associated with muscle wasting and / or impaired renal function.

[0084] In another embodiment, the method is a method of preventing at least one disease or condition associated with muscle wasting and / or impaired renal function, the method comprising administering a composition comprising a urolithin, or a composition comprising a prophylactically effective amount of a urolithin or a pharmacologically acceptable salt thereof, to a subject at risk of at least one disease or condition associated with muscle wasting and / or impaired renal function.

[0085] In yet another embodiment, the muscle is the tibialis anterior and / or the quadriceps.

[0086] In one embodiment, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to increase urinary creatinine, and / or decrease the albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve muscle lean mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

[0087] In certain embodiments, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to increase urinary creatinine and improve lean muscle mass.

[0088] In another embodiment, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to improve lean mass of muscle and / or improve muscle fiber size and / or improve the level of at least one amino acid.

[0089] In further embodiments, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to improve the level of at least one metabolic product, and / or improve the level of at least one nucleotide, and / or improve the level of at least one nicotinamide adenine dinucleotide, and / or increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increase the level of succinate and / or malate, and / or increase the level of phosphocreatine.

[0090] In a second aspect, the disclosure provides a method for improving lean muscle mass in a subject in need thereof, the method comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving lean muscle mass in a subject in need thereof. In related embodiments, the muscle is the tibialis anterior and / or the quadriceps.

[0091] In a third aspect, the disclosure provides a method for increasing albumin reabsorption in a subject in need thereof. The method comprises administering to a subject in need thereof an effective amount of a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof for use in a method for increasing albumin reabsorption in a subject in need thereof. Increasing albumin reabsorption means that the amount of albumin in the urine is reduced.

[0092] In a fourth aspect, the disclosure provides a method for increasing urinary creatinine in a subject in need thereof. The method comprises administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for increasing urinary creatinine in a subject in need thereof.

[0093] In a fifth aspect, the disclosure provides a method for reducing the albumin / creatinine ratio in a subject in need thereof. The method comprises administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for reducing the albumin / creatinine ratio in a subject in need thereof.

[0094] In a sixth aspect, the disclosure provides a method for increasing urinary carnosine in a subject in need thereof. The method comprises administering to a subject in need thereof an effective amount of a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof for use in a method for increasing urinary carnosine in a subject in need thereof.

[0095] In a seventh aspect, the present disclosure provides a method for increasing urinary anserine in a subject in need thereof. The method comprises administering to a subject in need thereof an effective amount of a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof. In other words, the present disclosure provides a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof for use in a method for increasing urinary anserine in a subject in need thereof.

[0096] In an eighth aspect, the disclosure provides a method for increasing urinary S-adenosylmethionine in a subject in need thereof. The method comprises administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for increasing urinary S-adenosylmethionine in a subject in need thereof.

[0097] In a ninth aspect, the disclosure provides a method for improving femoral bone mass in a subject in need thereof. The method comprises administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving femoral bone mass in a subject in need thereof.

[0098] The present disclosure provides methods for improving and / or maintaining muscle mass in a subject in need thereof, comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In other words, the present disclosure provides compositions comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in methods for improving and / or maintaining muscle mass in a subject in need thereof. In particular, the amount of the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is effective to improve lean mass of muscle and / or improve muscle fiber size and / or improve the level of at least one amino acid.

[0099] In a tenth aspect, the disclosure provides a method for improving muscle fiber size in a subject in need thereof, the method comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving muscle fiber size in a subject in need thereof.

[0100] Reductions in muscle mass and fiber size can lead to muscle atrophy, and thus, in one embodiment, administering a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof, to a subject in need thereof reduces muscle atrophy.

[0101] In an eleventh aspect, the disclosure provides a method for improving the level of at least one amino acid in muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving the level of at least one amino acid in muscle of a subject in need thereof.

[0102] In one embodiment, the amino acids whose levels are increased are selected from the group consisting of isoleucine, methionine, lysine, tyrosine, proline, alanine and glycine, hi one embodiment, the amino acids whose levels are increased are isoleucine and / or methionine and / or lysine and / or tyrosine and / or proline and / or alanine and / or glycine.

[0103] The present disclosure provides a method for improving muscular endurance and / or efficiency in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof. In other words, the present disclosure provides a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof for use in a method for improving muscular endurance and / or efficiency in a subject in need thereof. In particular, the composition comprising a urolithin or the amount of a urolithin or a pharmacologically acceptable salt thereof is effective to improve the level of at least one metabolic product, and / or improve the level of at least one nucleotide, and / or improve the level of at least one nicotinamide adenine dinucleotide, and / or increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increase the level of succinate and / or malate, and / or increase the level of phosphocreatine.

[0104] The endurance and / or efficiency of a muscle can be improved by improving its metabolism.

[0105] Accordingly, in a twelfth aspect, the disclosure provides a method for improving the level of at least one metabolic product in muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving the level of at least one metabolic product in muscle of a subject in need thereof.

[0106] In any embodiment, improving the level of a metabolic product may also include increasing and / or decreasing the level of a particular metabolic product.

[0107] In one embodiment, the at least one metabolic product is selected from the group consisting of N,N-dimethylglycine, S-adenosylmethionine (SAM), N-acetyl-DL-serine, N-acetyl-L-arginine, and N-acetylglutamic acid.

[0108] In another embodiment, the metabolic product is N-acetyl-DL-serine and / or N-acetyl-L-arginine and / or N-acetylglutamic acid.

[0109] In a further embodiment, the metabolic product is N,N-dimethylglycine and / or S-adenosylmethionine (SAM).

[0110] The method for improving the level of at least one metabolic product may also increase the level of ketone bodies and / or molecules associated with ketone bodies. In one embodiment, the metabolic product is 2-hydroxybutyrate.

[0111] In another embodiment, the level of at least one metabolic product is improved by decreasing the level of trans-urocanic acid.

[0112] In a further embodiment, the level of at least one metabolite is improved by increasing the level of anserine and / or carnosine in muscle.

[0113] Anserine and carnosine are associated with muscle efficiency and endurance. In particular, carnosine functions as a physiological buffer, has antioxidant properties, influences enzyme regulation, and acts on sarcoplasmic reticulum calcium regulation. In one embodiment, administering to a subject in need thereof a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof improves muscle efficiency and endurance.

[0114] In a thirteenth aspect, the disclosure provides a method for improving the level of at least one nucleotide in muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving the level of at least one nucleotide in muscle of a subject in need thereof.

[0115] In one embodiment, the at least one nucleotide is selected from the group consisting of adenosine triphosphate (ATP), adenosine diphosphate (ADP), guanosine triphosphate (GTP), guanosine diphosphate (GDP), uridine triphosphate (UTP), cytidine triphosphate (CTP), and flavin adenine dinucleotide (FAD).

[0116] In another embodiment, the nucleotides are adenosine triphosphate (ATP) and / or adenosine diphosphate (ADP) and / or guanosine triphosphate (GTP) and / or guanosine diphosphate (GDP) and / or uridine triphosphate (UTP) and / or cytidine triphosphate (CTP) and / or flavin adenine dinucleotide (FAD).

[0117] Nucleotides are a source of energy biomolecules and bioenergetics. In addition, nucleotides are important in cell signaling, for example participating in the purinergic signaling pathway. Thus, in a further embodiment, administering a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof to a subject in need thereof increases nucleotides of muscle bioenergetics and nucleotides important for cell signaling.

[0118] In a fourteenth aspect, the disclosure provides a method for improving the level of at least one nicotinamide adenine dinucleotide in a muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for improving the level of at least one nicotinamide adenine dinucleotide in a muscle of a subject in need thereof.

[0119] In one embodiment, the nicotinamide adenine dinucleotide is an oxidized form (NAD+) or a reduced form (NADH) of nicotinamide adenine dinucleotide. Preferably, the adenine dinucleotide is an oxidized form (NAD+) of nicotinamide adenine dinucleotide.

[0120] In another embodiment, the nicotinamide adenine dinucleotide is nicotinamide adenine dinucleotide phosphate (NADP). In particular, the nicotinamide adenine dinucleotide is the oxidized form (NADP+) or the reduced form (NADPH) of nicotinamide adenine dinucleotide phosphate. Preferably, the adenine dinucleotide is the oxidized form (NAD+) of nicotinamide adenine dinucleotide phosphate.

[0121] In a fifteenth aspect, the disclosure provides a method for increasing glutathione (GSH) and / or glutathione disulfide (GSSG) levels in muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for increasing glutathione (GSH) and / or glutathione disulfide (GSSG) levels in muscle of a subject in need thereof.

[0122] In one embodiment, the level of glutathione (GSH) is increased. In another embodiment, the level of glutathione disulfide (GSSG) is increased.

[0123] In one embodiment, administering a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof, to a subject in need thereof improves antioxidant buffering capacity in muscle.

[0124] In a sixteenth aspect, the disclosure provides a method for increasing succinate and / or maleate levels in muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for increasing succinate and / or maleate levels in muscle of a subject in need thereof.

[0125] In one embodiment, the level of succinate in muscle is increased, hi another embodiment, the level of malate in muscle is increased.

[0126] Succinate and maleate are metabolites of the tricarboxylic acid cycle (TCA), also known as the citric acid cycle. The metabolites of TCA are important for bioenergetics. Thus, in one embodiment, administering a composition comprising a urolithin or an effective amount of a urolithin or a pharmacologically acceptable salt thereof to a subject in need thereof improves muscle bioenergetics.

[0127] In a seventeenth aspect, the disclosure provides a method for increasing the level of phosphocreatine in muscle of a subject in need thereof, comprising administering to a subject in need thereof an effective amount of a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof. In other words, the disclosure provides a composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, for use in a method for increasing the level of phosphocreatine in muscle of a subject in need thereof.

[0128] Phosphocreatine functions as a rapidly available store of high-energy phosphate, and thus, in one embodiment, administering a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof, to a subject in need thereof improves muscle energy utilization.

[0129] In one embodiment, the methods disclosed herein may also be effective in treating a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

[0130] In another embodiment, the disease or condition is selected from the group consisting of chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, and PEW. Preferably, the disease is chronic kidney disease.

[0131] In further embodiments, the methods disclosed herein may also be effective in treating early stages of chronic kidney disease and / or late stages of chronic kidney disease. Preferably, early stages of chronic kidney disease are stages 2 and 3. In certain embodiments, early stages of chronic kidney disease are stages 2. Preferably, late stages of chronic kidney disease are stages 4 and 5. In certain embodiments, late stages of chronic kidney disease are stage 4.

[0132] In one embodiment, the methods disclosed herein may also be effective in treating early stages of chronic kidney disease.

[0133] In another embodiment, the methods disclosed herein may also be effective in the later stages of chronic kidney disease.

[0134] In one embodiment, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to increase urinary creatinine, and / or decrease the albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve muscle lean mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

[0135] In certain embodiments, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to increase urinary creatinine and improve lean muscle mass.

[0136] In another embodiment, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to improve lean mass of muscle and / or improve muscle fiber size and / or improve the level of at least one amino acid.

[0137] In further embodiments, the composition comprising a urolithin, or the amount of a urolithin or a pharmacologically acceptable salt thereof, is effective to improve the level of at least one metabolic product, and / or improve the level of at least one nucleotide, and / or improve the level of at least one nicotinamide adenine dinucleotide, and / or increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increase the level of succinate and / or malate, and / or increase the level of phosphocreatine.

[0138] In one embodiment, the composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof of the methods disclosed herein is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof. In particular, the composition may include additional dietary supplements, such as minerals, vitamins, and further biologically active substances, such as N-acetylglucosamine or N-acetylmuramic acid.

[0139] Thus, in an eighteenth aspect, the present disclosure provides a nutritional composition for use in the treatment and / or prevention of a disease or condition associated with muscle wasting and / or impaired kidney function, the nutritional composition comprising a urolithin or a composition comprising a urolithin or a pharmacologically acceptable salt thereof.

[0140] In one embodiment, the nutritional composition contains an amount of urolithin effective to treat and / or prevent a disease or condition associated with muscle wasting and / or kidney dysfunction in a subject in need thereof, hi a related embodiment, the nutritional composition is formulated for oral administration.

[0141] In a nineteenth aspect, the disclosure provides a unit dosage form for use in the treatment and / or prevention of a disease or condition associated with muscle wasting and / or impaired kidney function, the unit dosage form comprising a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof.

[0142] In one embodiment, the unit dosage form contains an amount of urolithin effective to treat and / or prevent a disease or condition associated with muscle wasting and / or impaired kidney function in a subject in need thereof. In a related embodiment, the unit dosage form is formulated for enteral administration.

[0143] In one embodiment, the amount of nutritional composition and / or unit dosage form is effective to increase urinary creatinine, and / or decrease albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve muscle lean mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

[0144] In another embodiment, the amount of nutritional composition and / or unit dosage form is effective to improve lean muscle mass and / or improve muscle fiber size and / or improve the level of at least one amino acid.

[0145] In further embodiments, the amount of the nutritional composition and / or unit dosage form is effective to improve the level of at least one metabolic product, and / or improve the level of at least one nucleotide, and / or improve the level of at least one nicotinamide adenine dinucleotide, and / or increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increase the level of succinate and / or malate, and / or increase the level of phosphocreatine.

[0146] In certain embodiments, the amount of the nutritional composition and / or unit dosage form is effective to increase urinary creatinine and improve lean muscle mass.

[0147] In one embodiment, the nutritional composition and / or unit dosage form is selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low volume liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

[0148] Food products according to the present invention include, but are not limited to, breads, cakes, cookies, crackers, extruded snacks, potato products, rice products, corn products, wheat products, dairy products, yogurt, confectionery, hard candy, gummy candy, nutritional bars, breakfast cereals or beverages. Food products according to the present invention may also be fruit juices, smoothies, plant-based beverages such as soy milk, rice milk, or almond milk.

[0149] In further embodiments, the nutritional compositions and unit dosage forms disclosed herein may be administered to subjects at early stages of chronic kidney disease and / or at later stages of chronic kidney disease.

[0150] In another embodiment, the nutritional compositions and unit dosage forms disclosed herein can be administered to a subject at an early stage of chronic kidney disease.

[0151] In one embodiment, the nutritional compositions and unit dosage forms disclosed herein can be administered to a subject in the later stages of chronic kidney disease.

[0152] In another embodiment, the urolithin is administered in the form of a composition comprising a urolithin.

[0153] In further embodiments, a urolithin in pure form or a pharmacologically acceptable salt thereof is administered.

[0154] In one embodiment, the compositions disclosed herein further comprise at least one additive.

[0155] In another embodiment, the urolithin is micronized for more rapid dispersion or dissolution. If a micronized urolithin is used, it is preferably 50 More preferably, the urolithin or precursor thereof has a D of less than 75 μm, such as less than 50 μm, for example less than 25 μm, such as less than 20 μm, for example less than 10 μm.50 More preferably, the urolithin or precursor thereof has a D in the range of 0.5 to 50 μm, for example, 0.5 to 20 μm, for example, 0.5 to 10 μm, for example, 1.0 to 10 μm, for example, 1.5 to 7.5 μm, for example, 2.8 to 5.5 μm. 50 Preferably, the urolithin or precursor thereof has a D of less than 100 μm. 90 More preferably, the urolithin or precursor thereof has a D of less than 75 μm, such as less than 50 μm, for example less than 25 μm, such as less than 20 μm, for example less than 15 μm. 90 The urolithin or precursor thereof preferably has a size in the range of 5 to 100 μm, such as 5 to 50 μm, for example 5 to 20 μm, for example 7.5 to 15 μm, for example 8.2 to 16.0 μm. 90 has.

[0156] Preferably, the urolithin has a D in the range of 0.5 to 1.0 μm. 10 Preferably, the urolithin has a D in the range of 8.2 to 16.0 μm. 90 , D in the range of 2.8 to 5.5 μm 50 , and D in the range of 0.5 to 1.0 μm 10 has.

[0157] Micronization can be achieved by a method selected from the group consisting of compression force grinding, hammer mills, universal mills or pin mills, and jet mills such as spiral jet mills or fluidized bed jet mills. Jet mills are particularly preferred.

[0158] Urolithins are metabolic products of food-derived ellagic acid derivatives such as ellagitannins and are produced in the human intestine by gut bacteria.

[0159] Ellagitannins are a class of antioxidant polyphenols present in several fruits, especially pomegranates, strawberries, raspberries, and walnuts. Ellagitannins are very poorly absorbed, but are rapidly metabolized to urolithins by the gut microbiota in the large intestine.

[0160] Due to their superior absorption, urolithins are believed to be bioactive molecules that mediate the effects of ellagitannins. For example, urolithins have previously been shown to have anti-proliferative, antioxidant and anti-inflammatory properties.

[0161] Examples of urolithins include ellagic acid, urolithin A (3,8-dihydroxyurolithin), urolithin B (3-hydroxyurolithin), urolithin C (3,8,9-trihydroxyurolithin), urolithin D (3,4,8,9-tetrahydroxyurolithin), urolithin A glucuronide, and urolithin B glucuronide. Ellagic acid and urolithin A, urolithin B, urolithin C, and urolithin D have the following structures:

[0162] [Table 1]

[0163] The compositions of the invention may comprise urolithins as defined above extracted from or contained in several fruits, such as pomegranate extract, tamarind extract or shilajit (mumijo) extract, strawberry, raspberry and walnut, which typically provide at least one urolithin or at least a portion of its precursors. In general, the compositions of the invention may comprise any of such pomegranate extract, tamarind extract or shilajit extract, strawberry, raspberry and walnut, or more preferably, urolithins extracted from such pomegranate extract, tamarind extract or shilajit extract, strawberry, raspberry and walnut. Alternatively, or additionally, the compositions of the invention may comprise any of the urolithins defined herein in isolated form. The isolated form may be prepared based on natural sources, such as those identified above, or may be provided by chemical synthesis.

[0164] In one embodiment, the urolithin is selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D, glucuronidated forms thereof, methylated forms thereof, sulfated forms thereof, and mixtures thereof. The urolithin may be provided as an isolated urolithin, such as a urolithin isolated from a natural source, or a urolithin prepared by total synthesis. An isolated urolithin may be synthesized de novo.

[0165] In certain embodiments, the urolithin is urolithin A.

[0166] The urolithin may be administered in an amount of about 0.2 to 150 milligrams (mg) of urolithin per kilogram (kg) of the subject's body weight. Preferably, the urolithin is administered at a dose equal to or corresponding to 2 to 120 mg of urolithin per kg of the subject's body weight, more preferably 4 to 90 mg of urolithin per kg of the subject's body weight, particularly preferably 6 to 20 mg of urolithin per kg of the subject's body weight, and most preferably 16 mg of urolithin per kg of the subject's body weight.

[0167] In one embodiment, a urolithin is administered at a dose sufficient to achieve a peak serum level of at least 0.001 micromolar (μM), preferably at least 0.01 μM, more preferably at least 0.1 μM, and most preferably at least 1 μM, at least 5 μM, or at least 10 μM. In one embodiment, a urolithin or precursor thereof is administered at a dose sufficient to achieve a sustained serum level of at least 0.001 micromolar (μM), preferably at least 0.01 μM, more preferably at least 0.1 μM, and most preferably at least 1 μM, at least 5 μM, or at least 10 μM. Sustained serum levels can be measured using any suitable method, for example, high pressure liquid chromatography (HPLC) or HPLC-MS.

[0168] In some embodiments, the urolithin is from 0.1% to 80% by weight of the composition, such as from 0.1% to 60% by weight of the composition, such as from 0.25% to 50% by weight of the composition, such as from 0.5% to 50% by weight of the composition. When the composition is provided as part or all of a meal, the urolithin may be from 0.25 to 5% by weight of the composition, such as from 0.3 to 3% by weight of the composition. When the composition is provided as a single serving of a dietary supplement to a subject's general diet, the urolithin may be from 20% to 80% by weight of the composition, such as from 20% to 40% by weight of the composition, such as from 25% to 35% by weight of the composition.

[0169] Urolithin can be administered in an amount of about 12 mg / day to about 9 g / day, preferably about 12 mg / day to about 7 g / day, more preferably about 12 mg / day to about 5 g / day, and most preferably about 12 mg / day to about 3 g / day, for example, about 12 mg / day to about 900 mg / day, about 12 mg / day to about 700 mg / day, about 12 mg / day to about 500 mg / day, about 12 mg / day to about 250 mg / day, about 12 mg / day to about 100 mg / day, or about 12 mg / day to about 50 mg / day, or about 12 mg / day to about 20 mg / day, or about 12 mg / day to about 18 mg / day. Of course, the daily dose can be administered in smaller doses at various times throughout the day. In any case, however, the amount of compound and / or composition administered will vary depending on factors such as the solubility of the active ingredient, the formulation used, the condition of the subject (e.g., body weight), and / or the route of administration. For example, the daily doses of urolithins disclosed above are non-limiting and may vary in some embodiments, in particular, compositions comprising urolithins or urolithins or pharmacologically acceptable salts thereof disclosed herein may be utilized as foods for acute special medical purposes (FSMPs) and contain up to about 100 mg / day of urolithin.

[0170] In one embodiment, administration of a composition containing a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, during the remission phase may be carried out for at least about 2 or 3 months, preferably at least about 4 or 5 months, more preferably at least about 6 or 7 months, for example, about 2 to 60 months, 2 to 48 months, 2 to 36 months, 2 to 24 months, or 2 to 12 months, preferably, for example, about 4 to 60 months, 4 to 48 months, 4 to 36 months, 4 to 24 months, or 4 to 12 months.

[0171] In another embodiment, a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof is administered for about 1 to 2 weeks to 6 months, more preferably about 2 weeks to 4 months, more preferably about 3 weeks to 3 weeks, and most preferably about 4 weeks to 10 weeks to increase urinary creatinine, and / or decrease the albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve lean muscle mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

[0172] In a further embodiment, a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof is administered for 5 weeks to obtain an increase in urinary creatinine, and / or a decrease in the albumin / creatinine ratio, and / or an increase in albumin reabsorption, and / or an improvement in muscle lean mass.

[0173] In one embodiment, a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof is administered for 10 weeks to increase urinary creatinine, and / or decrease albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve lean muscle mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

[0174] In another embodiment, a composition comprising a urolithin, or the above-mentioned amount of a urolithin or a pharmacologically acceptable salt thereof, is administered for 10 weeks to improve lean mass of muscle, and / or to improve muscle fiber size, and / or to improve levels of at least one amino acid.

[0175] In further embodiments, the composition comprising a urolithin or the above amounts of a urolithin or a pharmacologically acceptable salt thereof is administered for 10 weeks to improve the level of at least one metabolic product, and / or to improve the level of at least one nucleotide, and / or to improve the level of at least one nicotinamide adenine dinucleotide, and / or to increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or to increase the level of succinate and / or malate, and / or to increase the level of phosphocreatine.

[0176] The compositions comprising a urolithin disclosed herein or a urolithin or a pharmacologically acceptable salt thereof can be administered in any of a variety of formulations for therapeutic purposes. More specifically, the pharmaceutical compositions can include suitable pharmacologically acceptable carriers or diluents and can be formulated as solid, semi-solid, liquid or gaseous formulations, such as tablets, capsules, powders, granules, ointments, liquids, suppositories, injections, inhalants, gels, microspheres, and aerosols. Thus, administration of a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof can be achieved in a variety of ways, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, transdermal, and intratracheal administration. The active ingredient urolithin may be systemic after administration or may be localized by the use of topical administration, intramural administration, or the use of an implant that acts to retain an effective dose at the site of implantation.

[0177] In one embodiment, the composition comprising a urolithin or administration of a urolithin or a pharmacologically acceptable salt thereof is administered enterally.

[0178] In oral formulations, the compositions comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof can be used alone or in combination with suitable additives for producing tablets, powders, granules, or capsules, for example, in combination with conventional additives such as lactose, mannitol, corn starch, or potato starch, in combination with binders such as microcrystalline cellulose, functional derivatives of cellulose, gum arabic, corn starch, or gelatin, in combination with disintegrating agents such as corn starch, potato starch, or sodium carboxymethylcellulose, in combination with lubricants such as talc or magnesium stearate; and, if desired, in combination with diluents, buffers, wetting agents, preservatives, and flavoring agents.

[0179] Compositions comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof can be utilized in aerosol formulations to be administered by inhalation. For example, compositions comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof can be formulated into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, and nitrogen.

[0180] Additionally, suppositories can be prepared by mixing a composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof with a variety of bases, such as emulsifying bases or water-soluble bases. A composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof can be administered rectally via a suppository. Suppositories can include vehicles that melt at body temperature but solidify at room temperature, such as cocoa butter, carbowax, and polyethylene glycol.

[0181] Unit dosage forms for oral or rectal administration, such as syrups, elixirs, and suspensions, may be provided, with each dosage unit, e.g., a teaspoon, tablespoon, tablet or suppository, containing a urolithin-containing composition or a defined amount of a urolithin or a pharmacologically acceptable salt thereof.

[0182] In another embodiment, the composition comprising a urolithin or a urolithin or a pharmacologically acceptable salt thereof is administered parenterally. Non-limiting examples of parenteral administration include intravenous, intramuscular, intraperitoneal, subcutaneous, intra-articular, intrasynovial, intraocular, intrathecal, topical, and inhalation. Thus, non-limiting examples of compositions comprising a urolithin or forms of a urolithin or a pharmacologically acceptable salt thereof include whole foods, processed foods, natural fruit juices, concentrates and extracts, injections, microcapsules, nanocapsules, liposomes, plasters, inhalation forms, nasal sprays, nasal drops, eye drops, sublingual tablets, and sustained release formulations.

[0183] Unit dosage forms for injection or intravenous administration may comprise a composition comprising a urolithin or a composition containing a urolithin or a pharmacologically acceptable salt thereof in a solution in sterile water, saline or another pharma- ceutically acceptable carrier, with each dosage unit, e.g., mL or L, containing a defined amount of the composition comprising a urolithin or a composition containing one or more of the urolithin or a pharmacologically acceptable salt thereof. EXAMPLES

[0184] Example 1 The following non-limiting examples discuss experimental data investigated to further support the methods and compositions disclosed herein.

[0185] The renal protective effect of urolithin A was evaluated in a mouse model of type 2 diabetes (db / db mice).

[0186] The diabetic model mouse used was the Diabetic Complications Consortium (DiabComp / AMDCC)-approved obese, insulin-resistant "type 2" db / db mouse model, which has been shown to have mitophagy defects in the kidneys.

[0187] To obtain a sizable cohort for testing groups in tandem, which is important for reliable mitochondrial function studies, mice were purchased from The Jackson Laboratory (Stock No: 000642). Male db / db mice aged 8-9 weeks (n=15 per group) were administered Urolithin A by gavage (50 mg / kg / day) for a period of 10 weeks. db / m (control) or db / db (diabetic) mice were randomly assigned to the following treatments:

[0188] [Table 2]

[0189] Fifteen db / m and 30 db / db mice obtained from the Jackson Laboratory were used. Mice had free access to tap water and standard rodent chow (Specialty Feeds, WA, Australia). All mice were housed in a room with a constant temperature of approximately 25°C and a 12-h day / night cycle.

[0190] 0.5% carboxymethylcellulose (CMC, Sigma#419303) was first prepared. Urolithin A (16.67 mg / mL) was prepared by dissolving in 0.5% CMC solution and kept stirring at room temperature. Urolithin A was administered at 50 mg / kg / day. The dose of Urolithin A administered to each mouse was calculated using the mouse's weight based on weekly weight measurements. Approximately 100 μL of 0.5% CMC was given daily to the mice in the vehicle group.

[0191] At baseline, midpoints, and 1 week prior to euthanasia, mice were housed in metabolic cages for 24 hours to collect urine and measure food and water intake. At baseline, midpoints, and 2 weeks prior to euthanasia, body composition analysis was performed by EchoMRI to measure fat mass and lean body mass. At baseline and at week 9, oral glucose tolerance tests were performed to assess glucose disposal capacity. Fasting plasma glucose was measured at baseline, 15 and 60 min after glucose treatment using a glucose colorimetric assay kit (Cayman, Ann Arbor, USA). Blood pressure measurements were performed at week 10.

[0192] All analyses were performed by ANOVA followed by post hoc pairwise analysis using Tukey's least significant difference procedure and correcting for multiple comparisons. A value of p<0.05 was considered statistically significant. A Z score of 2 was used to exclude outliers.

[0193] One week before harvesting, the mice were housed in metabolic cages for 24 hours, and urine and blood were collected from the submandibular vein. Urinary and plasma creatinine were measured using a Cobas Integra automated analyzer. Urinary albumin was measured by ELISA (Bethyl, USA). Blood pressure was measured by tail cuff plethysmography. As a result, the albumin / creatinine ratio was significantly decreased in the db / db+UA group (UA) compared with the db / db+V group (diabetes) after 5 weeks of treatment (Fig. 1a) and 10 weeks of treatment (Fig. 1b). In addition, the results show that urinary creatinine in UA-treated db / db mice was higher than that in the vehicle-treated group (Fig. 2).

[0194] Urinary albumin excretion during a 24-h period was measured by ELISA kit (Bethyl). At the end of the study, there was a significant increase in albuminuria in db / db mice compared to db / m mice. Treatment with UA affected albuminuria in db / db mice at intermediate time points. Creatinine clearance was calculated from urinary and plasma creatinine measured using the Creatinine plus ver.2 (CREP2) assay on a Cobas Integra 400 plus autoanalyzer (Roche Diagnostics). In db / db mice (diabetic), there was an increase in albuminuria and a decrease in renal function assessed by a decrease in creatinine clearance at 18-19 weeks of age. Treatment with UA delayed the progression of albuminuria in db / db mice. The results show that the progression of albuminuria in UA-treated db / db (UA) mice was significantly delayed compared to the vehicle-treated counterparts (Figure 3).

[0195] Whole body composition, including lean mass and fat mass, was measured using EchoMRI at baseline, intermediate time points, and endpoint. Overall, db / db mice (diabetes) had increased fat mass and total body weight and decreased lean mass compared to db / m (control), regardless of treatment. Lean mass was significantly increased in the db / db+UA (UA) group compared to the db / db+V group (diabetes). The results are shown in Figure 4. In particular, the tibialis anterior and quadriceps muscles of the db / db+UA (UA) group show significantly increased muscle mass compared to the db / db+V group (diabetes). The results are shown in Figures 5a and 5b.

[0196] Urinary levels of carnosine, anserine and S-adenosylmethionine (SAM) were semi-quantitatively measured using HILIC chromatography and ESI-Orbitrap mass spectrometry. The levels of all these metabolites were decreased in the db / db+V group (diabetes) compared to the db / m group (healthy subjects) (Figures 6 to 8). Urinary carnosine levels were significantly increased in the db / db+UA (UA) group compared to the db / db+V (diabetes) group. The results are shown in Figure 6. Urinary anserine levels were significantly increased in the db / db+UA (UA) group compared to the db / db+V (diabetes) group. The results are shown in Figure 7. Urinary S-adenosylmethionine (SAM) levels were significantly increased in the db / db+UA (UA) group compared to the db / db+V (diabetes) group. The results are shown in Figure 8.

[0197] Left and right femurs were harvested at the end of the study after 10 weeks of treatment. Both femurs were washed, flash frozen, weighed, and stored at -80°C. Femoral mass was decreased in db / db+V mice compared to the db / m group (control), but significantly increased in the db / db+UA group (UA) compared to the db / db+V group (diabetes). The results are shown in Figure 9.

[0198] The size distribution of fibers measured within the tibia in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) was compared to the vehicle group (diabetic, db / db+V group) and the control group (control, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose.

[0199] Tibialis anterior, EDL, femoral, soleus and quadriceps muscles from the Monash study were harvested, weighed and stored frozen.

[0200] Fiber size distribution was assessed after tibialis anterior muscle cryosections and stained for laminin protein and myonuclei. All slides were acquired on an Olympus VS120 slide scanner microscope. Myofiber size was calculated in Min Feret using an automated image processing algorithm developed in-house using QuPath software and the Fiji tool open-CSAM. Results are shown in Figure 10.

[0201] Metabolites in muscle tissue were measured in mice after 10 weeks of treatment with Urolithin A at a dose of 50 mg / kg / day (+UA, db / db+UA group) and compared to the vehicle group (diabetic, db / db+V group) and control group (healthy, db / m+V group) after 10 weeks of treatment with 100 μL of 0.5% carboxymethylcellulose.

[0202] Muscle tissue extraction was performed in a tissue mixer (Qiagen TissueLyser II) with metal beads in a pre-cooled rack (-80°C) at 23 Hz for 2 min.

[0203] All samples were agitated for 10 min at 1500 rpm and 4° C. in a shaker (Thermomixer C, Eppendorf) followed by centrifugation for 10 min at 15,000 rpm and 4° C. Of note, for experiments without isotope-labeled substrate, a labeled internal standard (fully labeled 1000 μg / ml) was added during extraction for later data normalization. 13 C The samples were analyzed using a centrifugation step, which included yeast extract and nicotinamide-D4). After the centrifugation step, two phases were obtained: an upper phase containing polar metabolites and a lower phase containing non-polar metabolites. Furthermore, a protein layer remained in between the two phases. The upper phase was dried overnight in a vacuum centrifuge at 4° C. and 5 mbar and dissolved in 60 μL of 60% (v / v) acetonitrile:water before analysis. The protein layer of the muscle samples was quantified by bicinchoninic acid (BCA) assay (ThermoFisher Scientific) and used for subsequent normalization of metabolite concentrations.

[0204] 2 μL of each sample was injected onto a hydrophilic interaction chromatography (HILIC) analytical column (2.1 mm × 150 mm, 5 μm pore size, 200 Å HILICON iHILIC®-Fusion(P)) protected by a precolumn (2.1 mm × 20 mm, 200 Å HILICON iHILIC® Fusion(P) Guard Kit) operated at 35 °C. Separation was achieved by applying a linear solvent gradient. Solvent A as the mobile phase was 10 mM ammonium acetate (NH4Ac) and 0.04% (v / v) ammonium hydroxide (NH4OH) in H2O (pH ∼9.3), and solvent B was acetonitrile (ACN).

[0205] The sheath gas was at 20 AU and the auxiliary gas was maintained at 15 AU. The vaporizer temperature was 280 °C and the ion transport tube temperature was 310 °C. Full scans were measured with positive and negative mode scans alternating on the fly, covering the m / z ranges of 83–830 and 73–900, respectively, with a resolution of 60,000. Eluting metabolites were analyzed with an Orbitrap mass spectrometer (Orbitrap Fusion Lumos Tribrid, Thermo Scientific) equipped with a heated electrospray ionization (H-ESI) source. The software Xcalibur v4.1.31.9 (Thermo Scientific) was used for instrument control and data processing of isotopically labeled and unlabeled metabolites. The results for isoleucine, methionine, lysine, tyrosine, proline, alanine, glycine, N-acetyl-DL-serine, N-acetyl-L-arginine, N-acetylglutamic acid, N,N-dimethylglycine, SAM, 2-hydroxybutyrate, trans-urocanic acid, anserine, carnosine, ATP, ADP, GTP, GDP, UTP, CTP, FAD, NAD+, NADP+, GSH, GSSG, succinate, maleate and phosphocreatine are shown in Figures 11 to 20.

Claims

1. A method for treating and / or preventing a disease or condition associated with muscle wasting and / or impaired kidney function comprising administering to a subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

2. 2. The method of claim 1, wherein the disease or condition is selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, protein-energy wasting (PEW), and combinations thereof.

3. 2. The method of claim 1, wherein said muscle wasting and / or said renal dysfunction is treated and / or prevented in early stages of chronic kidney disease and / or in late stages of chronic kidney disease.

4. The method of claim 1 , wherein the muscle is the tibialis anterior and / or the quadriceps.

5. The method of claim 1, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

6. The method of claim 1, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

7. 2. The method of claim 1, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

8. 2. The method of claim 1, wherein the urolithin is micronized.

9. 2. The method of claim 1, wherein the urolithin is urolithin A.

10. 1. A method for improving lean muscle mass in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

11. 11. The method of claim 10, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

12. 11. The method of claim 10, wherein the lean mass of muscle is improved in early stages of chronic kidney disease and / or in late stages of chronic kidney disease.

13. The method of claim 10, wherein the muscle is the tibialis anterior and / or the quadriceps.

14. The method of claim 10, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

15. The method of claim 10, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

16. 11. The method of claim 10, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

17. 11. The method of claim 10, wherein the urolithin is micronized.

18. The method of claim 10, wherein the urolithin is urolithin A.

19. 1. A method for increasing albumin reabsorption in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

20. 20. The method of claim 19, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

21. 20. The method of claim 19, wherein the albumin reabsorption is increased in early stages of chronic renal disease and / or in late stages of chronic renal disease.

22. 20. The method of claim 19, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

23. 20. The method of claim 19, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

24. 20. The method of claim 19, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

25. 20. The method of claim 19, wherein the urolithin is micronized.

26. 20. The method of claim 19, wherein the urolithin is urolithin A.

27. 1. A method for increasing urinary creatinine in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

28. 28. The method of claim 27, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

29. 28. The method of claim 27, wherein the urinary creatinine is increased in early stages of chronic renal disease and / or in late stages of chronic renal disease.

30. The method of claim 27, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

31. The method of claim 27, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

32. 28. The method of claim 27, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

33. 28. The method of claim 27, wherein the urolithin is micronized.

34. 28. The method of claim 27, wherein the urolithin is urolithin A.

35. 1. A method for reducing albumin / creatinine ratio in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

36. 36. The method of claim 35, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

37. 36. The method of claim 35, wherein the albumin / creatinine ratio is decreased in early stages of chronic renal disease and / or in late stages of chronic renal disease.

38. The method of claim 35, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

39. The method of claim 35, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

40. 36. The method of claim 35, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

41. 36. The method of claim 35, wherein the urolithin is micronized.

42. The method of claim 35, wherein the urolithin is urolithin A.

43. 1. A method for increasing urinary carnosine in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

44. 44. The method of claim 43, wherein the subject has a disease or condition selected from chronic kidney disease, metabolically associated muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

45. 44. The method of claim 43, wherein the urinary carnosine is increased in early stages of chronic renal disease and / or in late stages of chronic renal disease.

46. The method of claim 43, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

47. The method of claim 43, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

48. 44. The method of claim 43, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

49. 44. The method of claim 43, wherein the urolithin is micronized.

50. The method of claim 43, wherein the urolithin is urolithin A.

51. 1. A method for increasing urinary anserine in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

52. 52. The method of claim 51, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

53. 52. The method of claim 51, wherein the urinary anserine is increased in early chronic renal disease and / or late chronic renal disease.

54. The method of claim 51, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

55. The method of claim 51, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

56. 52. The method of claim 51, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

57. 52. The method of claim 51, wherein the urolithin is micronized.

58. 52. The method of claim 51, wherein the urolithin is urolithin A.

59. 1. A method for increasing urinary S-adenosylmethionine in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

60. 60. The method of claim 59, wherein the subject has a disease or condition selected from chronic kidney disease, metabolically associated muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

61. 60. The method of claim 59, wherein the urinary S-adenosylmethionine is increased in early stages of chronic renal disease and / or in late stages of chronic renal disease.

62. 60. The method of claim 59, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

63. 60. The method of claim 59, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

64. 60. The method of claim 59, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

65. 60. The method of claim 59, wherein the urolithin is micronized.

66. 60. The method of claim 59, wherein the urolithin is urolithin A.

67. 1. A method for improving femoral bone mass in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

68. 68. The method of claim 67, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

69. 68. The method of claim 67, wherein the femoral mass is improved in early stages of chronic renal disease and / or in late stages of chronic renal disease.

70. The method of claim 67, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

71. The method of claim 67, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

72. 68. The method of claim 67, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

73. 68. The method of claim 67, wherein the urolithin is micronized.

74. 68. The method of claim 67, wherein the urolithin is urolithin A.

75. 1. A method for improving muscle fiber size in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

76. 76. The method of claim 75, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

77. 76. The method of claim 75, wherein muscle atrophy is reduced.

78. The method of claim 75, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

79. The method of claim 75, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

80. 76. The method of claim 75, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

81. 76. The method of claim 75, wherein the urolithin is micronized.

82. The method of claim 75, wherein the urolithin is urolithin A.

83. 1. A method for improving the level of at least one amino acid in a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

84. 84. The method of claim 83, wherein the subject has a disease or condition selected from chronic kidney disease, metabolically associated muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

85. 84. The method of claim 83, wherein the at least one amino acid is selected from the group consisting of isoleucine, methionine, lysine, tyrosine, proline, alanine, and glycine.

86. The method of claim 83, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

87. The method of claim 83, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

88. 84. The method of claim 83, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

89. 84. The method of claim 83, wherein the urolithin is micronized.

90. The method of claim 83, wherein the urolithin is urolithin A.

91. 1. A method for improving the level of at least one metabolic product in muscle of a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

92. 92. The method of claim 91, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

93. 92. The method of claim 91, wherein the at least one metabolic product is selected from the group consisting of N,N-dimethylglycine, S-adenosylmethionine (SAM), N-acetyl-DL-serine, N-acetyl-L-arginine, and N-acetylglutamic acid.

94. 92. The method of claim 91, wherein said at least one metabolic product is 2-hydroxybutyrate.

95. 92. The method of claim 91, wherein the level of the at least one metabolic product is improved by decreasing the level of trans-urocanic acid.

96. 92. The method of claim 91, wherein the level of at least one metabolic product is improved by increasing the level of anserine and / or carnosine in muscle.

97. 92. The method of claim 91, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

98. The method of claim 91, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

99. 92. The method of claim 91, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

100. 92. The method of claim 91, wherein the urolithin is micronized.

101. 92. The method of claim 91, wherein the urolithin is urolithin A.

102. 1. A method for improving levels of at least one nucleotide in muscle of a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

103. The method of claim 102, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

104. 103. The method of claim 102, wherein the at least one nucleotide is selected from the group consisting of adenosine triphosphate (ATP), adenosine diphosphate (ADP), guanosine triphosphate (GTP), guanosine diphosphate (GDP), uridine triphosphate (UTP), cytidine triphosphate (CTP), and flavin adenine dinucleotide (FAD).

105. The method of claim 102, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

106. The method of claim 102, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

107. 103. The method of claim 102, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

108. The method of claim 102, wherein the urolithin is micronized.

109. The method of claim 102, wherein the urolithin is urolithin A.

110. 1. A method for improving a level of at least one nicotinamide adenine dinucleotide in a muscle of a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

111. The method of claim 110, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

112. 111. The method of claim 110, wherein the at least one nicotinamide adenine dinucleotide is an oxidized form (NAD+) or a reduced form (NADH) of nicotinamide adenine dinucleotide.

113. 111. The method of claim 110, wherein the at least one nicotinamide adenine dinucleotide is an oxidized form (NADP+) or a reduced form (NADPH) of nicotinamide adenine dinucleotide phosphate.

114. The method of claim 110, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

115. The method of claim 110, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

116. 111. The method of claim 110, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

117. The method of claim 110, wherein the urolithin is micronized.

118. The method of claim 110, wherein the urolithin is urolithin A.

119. 1. A method for increasing glutathione (GSH) and / or glutathione disulfide (GSSG) levels in muscle of a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

120. 120. The method of claim 119, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

121. 120. The method of claim 119, wherein the antioxidant buffering capacity in the muscle is improved.

122. The method of claim 119, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

123. The method of claim 119, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

124. 120. The method of claim 119, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

125. The method of claim 119, wherein the urolithin is micronized.

126. The method of claim 119, wherein the urolithin is urolithin A.

127. 1. A method for increasing succinate and / or maleate levels in muscle of a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

128. 128. The method of claim 127, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

129. 128. The method of claim 127, wherein the muscle bioenergetics are improved.

130. The method of claim 127, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

131. The method of claim 127, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

132. 128. The method of claim 127, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

133. The method of claim 127, wherein the urolithin is micronized.

134. The method of claim 127, wherein the urolithin is urolithin A.

135. 1. A method for increasing the level of phosphocreatine in muscle of a subject in need thereof, comprising administering to the subject in need thereof a composition comprising a urolithin, or an effective amount of a urolithin or a pharmacologically acceptable salt thereof.

136. The method of claim 135, wherein the subject has a disease or condition selected from chronic kidney disease, metabolic muscle wasting, end stage renal disease, dialysis, diabetes, muscle weakness and / or renal failure due to intensive care unit admission, Pompe disease, metabolic acidosis, methylmalonic aciduria, disuse atrophy, PEW, and combinations thereof.

137. 136. The method of claim 135, wherein energy utilization in the muscle is improved.

138. The method of claim 135, wherein the composition comprising a urolithin, or a urolithin or a pharmacologically acceptable salt thereof, is administered enterally.

139. The method of claim 135, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered parenterally.

140. 136. The method of claim 135, wherein the composition comprising the urolithin, or the urolithin or a pharmacologically acceptable salt thereof, is administered as a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

141. The method of claim 135, wherein the urolithin is micronized.

142. The method of claim 135, wherein the urolithin is urolithin A.

143. A composition comprising a urolithin, or a nutritional composition comprising a urolithin or a pharmacologically acceptable salt thereof, for use in the treatment and / or prevention of a disease or condition associated with muscle wasting and / or kidney dysfunction, preferably wherein the nutritional composition contains an amount of urolithin effective for treating and / or preventing a disease or condition associated with muscle wasting and / or kidney dysfunction in a subject in need thereof.

144. 144. The composition of claim 143, comprising an amount of a urolithin effective to increase urinary creatinine, and / or decrease the albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve lean muscle mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

145. 144. The composition of claim 143, comprising an amount of a urolithin effective to improve lean mass of muscle and / or improve muscle fiber size and / or improve the level of at least one amino acid.

146. 144. The composition of claim 143, comprising an amount of a urolithin effective to improve the level of at least one metabolic product, and / or improve the level of at least one nucleotide, and / or improve the level of at least one nicotinamide adenine dinucleotide, and / or increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increase the level of succinate and / or malate, and / or increase the level of phosphocreatine.

147. 144. The composition of claim 143, wherein the muscle is the tibialis anterior and / or the quadriceps.

148. 144. The composition of claim 143, selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

149. 144. The composition of claim 143, formulated for oral administration.

150. The composition of claim 143, wherein the urolithin is micronized.

151. The composition of claim 143, wherein the urolithin is urolithin A.

152. A composition comprising a urolithin or a unit dosage form comprising a urolithin or a pharmacologically acceptable salt thereof for use in the treatment and / or prevention of a disease or condition associated with muscle wasting and / or renal dysfunction, preferably said unit dosage form containing an amount of urolithin effective for treating and / or preventing a disease or condition associated with muscle wasting and / or renal dysfunction in a subject in need thereof. Unit dosage form.

153. 153. The unit dosage form of claim 152, comprising an amount of a urolithin effective to increase urinary creatinine, and / or decrease the albumin / creatinine ratio, and / or increase albumin reabsorption, and / or improve lean muscle mass, and / or increase urinary carnosine, and / or increase urinary anserine, and / or increase urinary S-adenosylmethionine, and / or improve femoral bone mass.

154. 153. The unit dosage form of claim 152, containing a urolithin in an amount effective to improve lean muscle mass and / or improve muscle fiber size and / or improve the level of at least one amino acid.

155. 153. The unit dosage form of claim 152, containing an amount of a urolithin effective to improve the level of at least one metabolic product, and / or improve the level of at least one nucleotide, and / or improve the level of at least one nicotinamide adenine dinucleotide, and / or increase the level of glutathione (GSH) and / or glutathione disulfide (GSSG), and / or increase the level of succinate and / or malate, and / or increase the level of phosphocreatine.

156. 153. The unit dosage form of claim 152, wherein the muscle is the tibialis anterior and / or the quadriceps.

157. 153. The unit dosage form of claim 152, selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a dairy-based beverage, a low-dose liquid nutritional supplement, a meal replacement beverage, and combinations thereof.

158. 153. The unit dosage form of claim 152, wherein the unit dosage form is formulated for enteral administration.

159. The unit dosage form of claim 152, wherein the urolithin is micronized.

160. The unit dosage form of claim 152, wherein the urolithin is urolithin A.

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