Risk prediction and stratification methods for sarcopenia and NAD deficiency

Abstract

The present invention relates generally to the field of muscle diseases. In particular, the present invention relates to methods for determining whether an individual suffers from sarcopenia or has an increased risk of having sarcopenia. Methods for predicting the response of the individual to nutritional intervention are also provided.

Description

Background Art

[0001] Age-related loss of muscle mass and function is inevitable in all individuals; however, its progression is largely dependent on genetic and environmental factors, such as physical activity and nutritional intake.

[0002] Sarcopenia has been defined as the point at which age-related loss of muscle mass and function renders the patient frail and affects quality of life. In contrast, frailty is another classification of age-related decline in physical function that is characterized by low muscle strength and low muscle function rather than muscle mass. Sarcopenia is clinically defined based on low muscle mass and low muscle function using cutoffs that stratify the elderly population for individuals with pathological activity. Sarcopenia predicts future disability and death and was designated an official ICD-10 disease code in 2016 (Anker et al., 2016).

[0003] Sarcopenia is common in men and women over the age of 65 years, with a global prevalence estimated at 3% to 30%, depending on the operational definition used. Sarcopenia is associated with numerous adverse physical and metabolic outcomes, including frailty, disability, obesity, diabetes, and osteoporosis. As a result, sarcopenia contributes to substantial health care costs. Although age, sex, body size, heritability, and physical activity are considered as influencing factors in adults, there remains considerable unexplained variability in muscle mass and strength in older individuals.

[0004] Sarcopenias are multifactorial syndromes associated with pathophysiological changes such as impaired neuromuscular transformation, altered excitation / contraction coupling, impaired regenerative capacity linked to stem cell exhaustion, defects in mitochondrial and energy metabolism in myofibers, and ultimately skeletal muscle marbling with fat and fibrosis. The etiology of these syndromes is therefore complex and poorly understood, but low physical activity, hormonal decline in anabolic hormones (androgens, IGF-1), and malnutrition / deficiency play an important role.

[0005] The MEMOSA (Multi-Ethnic Molecular Determinants of Human Sarcopenia) study was designed to identify molecular and nutritional changes associated with human sarcopenia.

[0006] The inventors of the present patent application have discovered that anthranilic acid and trigonelline levels in the body, as well as the ratio of trigonelline:anthranilic acid levels, are indicative of an individual suffering from sarcopenia or having an increased risk of developing sarcopenia. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1Serum levels of trigonelline vary in sarcopenic patients and are positively correlated with lean muscle mass, grip strength, and gait.

[0008] Figure 2 , serum levels of anthranilic acid differ in sarcopenic patients and are negatively correlated with lean muscle mass, grip strength, and gait.

[0009] Figure 3 Serum levels of trigonelline:anthranilic acid ratios differ in sarcopenic patients and are positively correlated with lean muscle mass, grip strength, and gait.

[0010] Figure 4A and Figure 4B . Figure 4A is a gene set enrichment analysis of sarcopenic versus control muscle, and Figure 4B Is selected from Figure 4A GSEA enrichment curves for the oxidative phosphorylation gene set. RNA sequencing of human sarcopenic skeletal muscle reveals mitochondrial dysfunction as a major transcriptional change during sarcopenia.

[0011] Figure 5A and Figure 5B . Figure 5A is a gene set enrichment analysis of sarcopenic versus control muscle, and Figure 5B Is selected from Figure 5A GSEA enrichment curve of oxidative phosphorylation gene set. Positive correlation between skeletal muscle mitochondrial function gene expression and serum level of trigonelline.

[0012] Fig. 6A and Figure 6B . Fig. 6A is a gene set enrichment analysis of sarcopenic versus control muscle, and Figure 6B Is selected from Fig. 6A GSEA enrichment curve of oxidative phosphorylation gene set. Negative correlation between skeletal muscle mitochondrial function gene expression and serum levels of anthranilic acid.

[0013] Fig. 7A and Figure 7B . Fig. 7A is a gene set enrichment analysis of sarcopenic versus control muscle, and Figure 7B Is selected from Fig. 7A GSEA enrichment curve of oxidative phosphorylation gene set. Positive correlation between skeletal muscle mitochondrial function gene expression and serum level ratio of trigonelline: anthranilic acid.

[0014] Figure 8 Human sarcopenic muscle has lower expression of genes such as NMAT1, NAMPT, and PNP.

[0015] Fig. 9 .NAD+ levels in muscle biopsies from control and sarcopenic participants.

[0016] Fig.10 .The ratio of serum levels of trigonelline:anthranilic acid was positively correlated with lean muscle mass index. DETAILED DESCRIPTION

[0017] The present invention relates to a method for determining whether an individual suffers from sarcopenia or has an increased risk of developing sarcopenia, the method comprising:

[0018] (a) determining the level of anthranilic acid in a sample obtained from the individual;

[0019] (b) comparing the level of anthranilic acid in the sample with a reference value;

[0020] Wherein an increased level of anthranilic acid in the sample compared to the reference value is an indication of sarcopenia or a risk of developing sarcopenia.

[0021] In one embodiment, the method further comprises:

[0022] (a) determining the level of trigonelline in a sample obtained from the individual;

[0023] (b) comparing the level of trigonelline in the sample with a reference value;

[0024] Wherein a reduced level of trigonelline in the sample compared to the reference value is an indication that the individual suffers from sarcopenia or is at risk of developing sarcopenia.

[0025] The present invention also relates to a method for determining whether an individual has sarcopenia or has an increased risk of developing sarcopenia, the method comprising:

[0026] (a) determining the level of trigonelline and the level of anthranilic acid in a sample obtained from the individual;

[0027] (b) determining the ratio of trigonelline: anthranilic acid in the sample;

[0028] (c) comparing the ratio of trigonelline: anthranilic acid in the sample with the reference value;

[0029] Wherein the ratio of trigonelline:anthranilic acid in the sample compared to a reference value is an indication that the individual suffers from or is at risk of developing sarcopenia.

[0030] In one embodiment, a decrease in the ratio of trigonelline:anthranilic acid in a sample from an individual compared to a reference value is an indication that the individual suffers from sarcopenia or has an increased risk of developing sarcopenia.

[0031] The present invention also relates to a method for determining whether an individual has sarcopenia or has an increased risk of developing sarcopenia, the method comprising:

[0032] (a) determining the level of trigonelline and the level of anthranilic acid in a sample obtained from the individual;

[0033] (b) determining the ratio of anthranilic acid:trigonelline in the sample;

[0034] (c) comparing the ratio of anthranilic acid:trigonelline in the sample with the reference value;

[0035] The ratio of anthranilic acid:trigonelline in the sample compared to a reference value is an indication that the individual suffers from or is at risk of developing sarcopenia.

[0036] In one embodiment, an increase in the ratio of anthranilic acid:trigonelline in a sample from an individual compared to a reference value is an indication that the individual suffers from sarcopenia or has an increased risk of developing sarcopenia.

[0037] The present invention also relates to a method for determining whether an individual has sarcopenia or has an increased risk of developing sarcopenia, the method comprising:

[0038] (a) determining the level of trigonelline and the level of anthranilic acid in a sample obtained from the individual;

[0039] (b) determining the relative amount or combination of anthranilic acid and trigonelline in a sample;

[0040] (c) comparing the relative amounts or combinations of anthranilic acid and trigonelline in the sample with reference values;

[0041] The relative amounts or combination of anthranilic acid and trigonelline in the sample compared to a reference value is indicative that the individual suffers from or is at risk of developing sarcopenia.

[0042] and wherein the combination is defined as n*f(A)+m*g(B), where A and B are the concentrations of trigonelline and anthranilic acid, f() and g() represent mathematical transformations, and n and m are numerical coefficients that may be positive or negative.

[0043] For example, the combination may be defined as 1*log2(A)+(-1)*log2(B)=log2(A / B) or 1*log2(A)-2log2(B)=log2(A / (B)^2).

[0044] In one embodiment, the mathematical transformation comprises a logarithmic transformation.

[0045] In one embodiment, an increase in the relative amount of anthranilic acid to trigonelline in a sample from an individual compared to a reference value is an indication that the individual suffers from sarcopenia or has an increased risk of developing sarcopenia.

[0046] In one embodiment, the invention relates to a method of determining whether an individual suffers from sarcopenia.

[0047] In one embodiment, the invention relates to a method of determining whether an individual has an increased risk of developing sarcopenia.

[0048] Sarcopenia is characterized by one or more of low muscle mass, low muscle strength and low physical fitness. More preferably, sarcopenia is characterized by two or more of low muscle mass, low muscle strength and low physical fitness. Most preferably, sarcopenia is characterized by low muscle mass, low muscle strength and low physical fitness. These can all be measured by methods well known to those skilled in the art.

[0049] Muscle mass can be measured by CT (computed tomography), DXA (dual-energy X-ray absorptiometry), MRI (magnetic resonance imaging), or D3 creatine dilution.

[0050] Muscle strength can be measured by grip strength (eg, using a handheld dynamometer) or knee extensor strength (eg, using quadriceps torque measurement).

[0051] Physical fitness can be measured by walking speed, SPPB, 400-m walk test, timed up and go test, or stair climbing test.

[0052] In one embodiment, the level of trigonelline and the level of anthranilic acid are determined by mass spectrometry.Preferably, the level of trigonelline and the level of anthranilic acid are determined by liquid chromatography followed by mass spectrometry.

[0053] In one embodiment, the subject is a human subject.

[0054] In one embodiment, the human subject is an elderly person. In one embodiment, the human subject is an elderly person.

[0055] In one embodiment, the subject is a companion animal, preferably a dog.

[0056] The present invention relates to a method for determining whether an individual suffers from a muscle wasting disorder or a muscle disease or has an increased risk of developing a muscle wasting disorder or a muscle disease, the method comprising:

[0057] (a) determining the level of anthranilic acid and / or the level of trigonelline in a sample obtained from an individual;

[0058] (b) comparing the level of anthranilic acid and / or the level of trigonelline in the sample with a reference value;

[0059] An increased level of anthranilic acid in the sample compared to a reference value or a decreased level of trigonelline in the sample compared to a reference value is indicative of a muscle wasting disorder or a muscle disease or a risk of developing a muscle wasting disorder or a muscle disease.

[0060] The present invention also relates to a method for determining whether an individual suffers from a muscle wasting disorder or a muscle disease or has an increased risk of developing a muscle wasting disorder or a muscle disease, the method comprising:

[0061] (a) determining the level of trigonelline and the level of anthranilic acid in a sample obtained from the individual;

[0062] (b) determining the ratio of trigonelline: anthranilic acid in the sample;

[0063] (c) comparing the ratio of trigonelline: anthranilic acid in the sample with the reference value;

[0064] Wherein the ratio of trigonelline:anthranilic acid in the sample compared to the reference value is an indication that the individual suffers from a muscle wasting disorder or a muscle disease or has an increased risk of developing a muscle wasting disorder or a muscle disease.

[0065] In one embodiment, a decrease in the ratio of trigonelline:anthranilic acid in a sample from an individual compared to a reference value is an indication that the individual has a muscle wasting disorder or a muscle disease or has an increased risk of developing a muscle wasting disorder or a muscle disease.

[0066] Muscle wasting symptoms may result from disuse, immobilization, prolonged bed rest, ICU (intensive care unit) hospitalization.

[0067] Muscle diseases can be sarcopenia, cachexia (caused by chronic diseases like cancer, COPD, heart failure, kidney disease), genetic myopathies and dystrophies such as Duchenne muscular dystrophy induced by drugs like corticosteroids or statins, muscle toxicity or rhabdomyolysis.

[0068] Preferably, the muscle disease is sarcopenia.

[0069] The following embodiments relate to one or more of the above-described methods of the present invention.

[0070] In one embodiment, the reference value is determined from a sample obtained from the same individual or from a group of individuals.

[0071] In one embodiment, the reference value is measured in a sample from the same individual.

[0072] In one embodiment, the reference value is measured in the same individual before a nutritional intervention (eg, use of a nutritional composition) and is compared to a measurement after the nutritional intervention.

[0073] In one embodiment, the reference value is measured in samples from a group of individuals (eg, a group of individuals of similar age).

[0074] In one embodiment, the subject is an elderly person.

[0075] In one embodiment, the subject is an elderly person.

[0076] In one embodiment, the sample obtained from the individual is a plasma sample.

[0077] In one embodiment, the sample obtained from the individual is a serum sample.

[0078] In one embodiment, the sample obtained from the individual is a urine sample.

[0079] The present invention also relates to a nutritional composition for treating or preventing sarcopenia in an individual.

[0080] In one embodiment, the individual is identified as having sarcopenia or as being at increased risk for developing sarcopenia.

[0081] In one embodiment, by a method according to the invention, an individual is identified as suffering from sarcopenia or as being at increased risk of developing sarcopenia.

[0082] In one embodiment, the nutritional composition comprises a NAD precursor. In one embodiment, the nutritional composition comprises trigonelline. In one embodiment, the nutritional composition comprises a precursor of trigonelline. In one embodiment, the nutritional composition comprises a derivative of trigonelline. Trigonelline is an alkaloid with the chemical formula C7H7NO2 and the CAS number 535-83-1.

[0083] In one embodiment, the subject is an elderly person.

[0084] In one embodiment, the subject is an elderly person.

[0085] In one embodiment, the individual is identified as having sarcopenia by the methods according to the invention, the nutritional composition comprises a NAD precursor, and the individual is an elderly person.

[0086] In one embodiment, the individual is identified as having sarcopenia by the methods according to the invention, the nutritional composition comprises trigonelline, and the individual is an elderly person.

[0087] The present invention also relates to methods for treating or preventing sarcopenia in a subject.

[0088] In one embodiment, the individual is identified as having sarcopenia or as being at increased risk for developing sarcopenia.

[0089] In one embodiment, an individual is identified as suffering from sarcopenia or as being at increased risk of developing sarcopenia by a method according to the invention.

[0090] In one embodiment, the method comprises modifying the individual's lifestyle.

[0091] In one embodiment, lifestyle modifications include physical activity, such as weight-bearing, resistance, or endurance exercise.

[0092] In one embodiment, modifying an individual's lifestyle comprises a change in diet.

[0093] In one embodiment, the dietary changes include administering at least one nutritional composition to the individual as part of a diet to treat or prevent sarcopenia.

[0094] In one embodiment, the nutritional composition comprises a NAD precursor.

[0095] In one embodiment, the subject is an elderly person.

[0096] In one embodiment, the subject is an elderly person.

[0097] The present invention also relates to methods for predicting the response of an individual suffering from, or at increased risk of developing, muscle wasting or a muscle disease to a nutritional composition.

[0098] In one embodiment, the nutritional composition comprises a NAD precursor, the individual is an elderly person, and the muscle disease is sarcopenia.

[0099] The present invention also relates to methods for predicting the response of an individual suffering from sarcopenia or at increased risk of developing sarcopenia to a nutritional composition.

[0100] In one embodiment, the nutritional composition comprises a NAD precursor and the individual is an elderly person.

[0101] In one embodiment, the method of the present invention comprises:

[0102] (a) detecting the level of one or more biomarkers in a sample obtained from the individual;

[0103] (b) comparing the level of one or more biomarkers in the sample with a reference value;

[0104] (c) predicting a response of the individual to the nutritional composition based on the results of step (b);

[0105] Wherein the one or more biomarkers are anthranilic acid and / or trigonelline.

[0106] If the level of anthranilic acid in the sample is increased compared to the reference value, it will be predicted that the individual will respond to the nutritional composition or nutritional intervention. If the level of trigonelline in the sample is decreased compared to the reference value, it will be predicted that the individual will respond to the nutritional composition or nutritional intervention.

[0107] In one embodiment, the biomarker is anthranilic acid.

[0108] In one embodiment, the biomarker is trigonelline.

[0109] In one embodiment, the biomarker is a combination of anthranilic acid and trigonelline.

[0110] In one embodiment, the biomarker is anthranilic acid, trigonelline, 3-hydroxy-anthranilic acid, tryptophan, kynurenine, or a metabolite thereof.

[0111] In one embodiment, the nutritional composition is vitamin B3.

[0112] In one embodiment, the nutritional composition is a NAD precursor, such as nicotinamide riboside, nicotinamide mononucleotide, and nicotinamide.

[0113] In one embodiment, the nutritional composition is tryptophan, kynurenine, and metabolites thereof.

[0114] In one embodiment, the nutritional composition is trigonelline.

[0115] In one embodiment, the nutritional composition is derived from coffee, such as a coffee bean extract or a coffee beverage. In one embodiment, the coffee extract comprises one or more of caffeic acid, trigonelline, quinolinic acid, and a precursor of quinolinic acid.

[0116] individual

[0117] The term "subject" refers to any animal, including humans and companion animals. Generally speaking, the subject is a human or an avian, bovine, canine, equine, feline, caprine, murine, ovine and porcine animal. The subject can be a horse or a companion animal, such as a cat or dog. Preferably, the subject is a human. In the context of humans, the term "elderly" refers to an age of at least 60 years old, more preferably over 64 years old, and most preferably over 68 years old since birth. In the context of humans, the term "middle-aged and elderly" refers to an age of at least 45 years old, preferably over 50 years old, more preferably over 55 years old since birth, and includes elderly individuals.

[0118] Sarcopenia

[0119] Sarcopenia can be characterized by one or more of low muscle mass, low muscle strength, and low physical fitness.

[0120] Sarcopenia in an individual can be diagnosed based on the definition of AWGSOP (Asian Working Group on Sarcopenia in Older Adults), for example as described by Chen et al. in 2014. Low muscle mass can generally be based on low appendicular lean mass (ALM index) normalized to the square of height, specifically an ALM index of less than 7.00 kg / m for men. 2 , and the ALM index of women is less than 5.40 kg / m 2 Low physical performance may be generally based on walking speed, specifically a walking speed of less than 0.8 m / sec. Low muscle strength may be generally based on low grip strength, specifically a grip strength of less than 26 kg for males and less than 18 kg for females.

[0121] Sarcopenia in an individual can be diagnosed based on the definition of EWGSOP (European Working Group on Sarcopenia in Older People), for example as described by Cruz-Jentoft et al. in 2010. Low muscle mass can generally be based on low limb lean mass (ALM index) normalized to the square of height, specifically an ALM index of less than 7.23 kg / m for men. 2 , and the ALM index of women is less than 5.67kg / m 2 Low physical performance may be generally based on walking speed, specifically a walking speed of less than 0.8 m / sec. Low muscle strength may be generally based on low grip strength, specifically a grip strength of less than 30 kg for males and less than 20 kg for females.

[0122] Sarcopenia can be diagnosed in an individual based on the definition of the Foundation for the National Institutes of Health (FNIH), for example, as described in Studenski et al., 2014. Low muscle mass can be generally based on low limb lean mass (ALM) normalized to body mass index (BMI; kg / m2), specifically ALM:BMI less than 0.789 for males and ALM:BMI less than 0.512 for females. Low physical performance can be generally based on walking speed, specifically walking speed less than 0.8 m / sec. Low muscle strength can be generally based on low grip strength, specifically grip strength less than 26 kg for males and grip strength less than 16 kg for females. Low muscle strength can also be generally based on low grip strength: body mass index, specifically grip strength: body mass index less than 1.00 for males and grip strength: body mass index less than 0.56 for females.

[0123] The D3-creatine dilution method is another method for measuring muscle mass. This method is becoming more widely accepted as a robust standard and is expected to replace DXA in the future. The D3-creatine dilution method has been previously described in Clark et al. (2014) and Stimpson et al. (2013).

[0124] The method of the present invention involves determining the level of trigonelline, the level of anthranilic acid and / or the ratio of their levels in a sample obtained from an individual. This may be referred to as a "test sample". Thus, the method of the present invention is typically performed in vitro in humans or animals, for example, on a body fluid sample previously obtained from the individual to be tested.

[0125] The sample may be, for example, a serum, plasma or urine sample. The sample may be derived from blood, i.e., the sample comprises whole blood or a blood fraction. The sample may comprise plasma or serum. Preferably, the sample is a serum sample.

[0126] Techniques for collecting blood samples and separating blood fractions are well known in the art. For example, a venous blood sample can be collected from a patient with a needle and deposited in a plastic tube. The collection tube may, for example, contain sprayed silica and polymer gel for serum separation. The serum can be separated by centrifugation at 1300 RCF for 10 minutes at room temperature and stored in small plastic tubes at -80°C.

[0127] Comparison with reference values

[0128] The method of the present invention involves comparison of anthranilic acid levels and trigonelline levels with reference value levels of anthranilic acid and trigonelline. The term "reference value" is synonymous with "control value" and broadly includes data that a person skilled in the art would use to facilitate accurate interpretation of technical data.

[0129] The level of anthranilic acid, the level of trigonelline and / or the ratio of trigonelline level:anthranilic acid level in a sample from an individual can be compared to the same item in the same individual or to the same item in one or more control groups of individuals.

[0130] Reference value can be established by measuring the sample from the same individual, for example, by repeating the mensuration of sample. Reference value can also be established in the following manner: measure the sample from the individual in the general population or selected population, and use statistical model, such as for selecting the positive standard or the predictive value method of the receiver operating characteristic curve that limits the best specificity (highest true negative rate) and sensitivity (highest true positive rate), as Knapp, RG and Miller, MC (1992), in " Clinical Epidemiology and Biostatistics " (" Clinical Epidemiology and Biostatistics ", William and Wilkins United Press (Harual Publishing Co.), Malvern, Pennsylvania, USA), described in, the document is incorporated herein by reference.

[0131] The reference value can be established by measuring at least 1 individual, more preferably at least 5 individuals, more preferably at least 10 individuals, more preferably at least 15 individuals, most preferably at least 20 individuals. All or most of the individuals preferably do not suffer from sarcopenia.

[0132] It is known in the art how to assign correct reference values, as they will vary with eg sex, ethnicity, genetic heritage, health status or age.

[0133] Nutritional composition

[0134] The nutritional composition according to the invention may be a nutraceutical composition, a functional food, a functional nutritional product, a medical food regulated as a drug, a medical nutritional product or a dietary supplement.

[0135] The term "nutraceutical" combines the words "nutrition" and "drug" together. It is a food or food product that provides health and medical benefits, including the prevention and treatment of disease. Nutrients are products separated or purified from foods that are generally sold in the form of drugs that are not usually associated with food. Nutrients are proven to have physiological beneficial effects or provide prevention of chronic diseases. Such products can range from isolated nutrients, dietary supplements, and special diets to genetically engineered foods, herbal products, and processed foods such as cereals, soups, and beverages.

[0136] As used herein, the term "nutraceutical" refers to usefulness in the fields of nutrition and pharmaceutical applications. Thus, nutraceutical compositions can be used as supplements to food and beverages, and can be used as pharmaceutical preparations for enteral or parenteral administration, which can be solid preparations such as capsules or tablets, or liquid preparations such as solutions or suspensions.

[0137] The nutritional composition according to the invention may comprise trigonelline, a precursor of trigonelline and / or a derivative of trigonelline. The precursor of trigonelline may be, for example, quinolinic acid, nicotinic acid and / or nicotinamide.

[0138] Preferably, the nutritional composition according to the invention comprises an NAD precursor, such as nicotinamide riboside, nicotinamide mononucleotide, nicotinamide and / or nicotinic acid.

[0139] The nutritional composition according to the invention may also comprise protective hydrophilic colloids (such as gums, proteins, modified starches), binders, film formers, encapsulating agents / materials, wall / shell materials, matrix compounds, coatings, emulsifiers, surfactants, solubilizers (oils, fats, waxes, lecithins, etc.), adsorbents, carriers, fillers, co-compounds, dispersants, wetting agents, processing aids (solvents), flow agents, taste masking agents, weighting agents, gelling agents, gel forming agents, antioxidants and antimicrobial agents.

[0140] In addition, multivitamin and mineral supplements can be added to the nutritional composition of the present invention to obtain sufficient amounts of essential nutrients that are lacking in some diets. Multivitamin and mineral supplements can also be used for disease prevention and to prevent nutritional losses and deficiencies due to lifestyle patterns.

[0141] The nutritional composition of the present invention may be in any galenic form suitable for administration to the body, in particular any form conventionally used for oral administration, for example in solid form, such as (additives / supplements to) food or feed, food or feed premixes, fortified food or feed, tablets, pills, granules, dragees, capsules and effervescent preparations (such as powders and tablets), or in liquid form, such as solutions, emulsions or suspensions, for example beverages, pastes and oily suspensions. The paste may be incorporated into a hard or soft shell capsule, whereby the capsule has a matrix of, for example, (fish, porcine, poultry, bovine) gelatin, vegetable proteins or lignin sulfonates. Other forms of administration are exemplified by forms for transdermal administration, parenteral administration or administration by injection. The nutritional composition may be in the form of a controlled (delayed) release formulation.

[0142] The nutritional composition may include beverages. The term "beverage" encompasses non-alcoholic and alcoholic beverages and liquid preparations to be added to drinking water and liquid foods. Non-alcoholic beverages are, for example, soft drinks, sports drinks, fruit juices, teas, and milk-based beverages. Liquid foods may include soups and dairy products. The nutritional composition comprising the compound of the present invention may be added to soft drinks, energy bars, or candies.

[0143] If the nutritional composition is a nutraceutical formulation, the composition further comprises a pharmaceutically acceptable excipient, diluent or adjuvant, which can then be formulated using standard techniques, such as disclosed in, for example, "Remington's Pharmaceutical Sciences" (Remington's Pharmaceutical Sciences, 20th edition, William & Wilkins Publishers, Pennsylvania, USA). For oral administration, tablets and capsules containing suitable binders (e.g., gelatin or polyvinyl pyrrolidone), suitable fillers (e.g., lactose or starch), suitable lubricants (e.g., magnesium stearate) and optionally other additives are preferably used.

[0144] “Functional foods,” “functional nutritional products,” “medical foods,” and “medical nutritional products” refer to any health food that claims to have health-promoting or disease-preventing properties beyond the basic function of providing nutrients. The general category of functional foods includes processed foods or foods fortified with additives that promote health, such as “vitamin-enriched” products.

[0145] Dietary supplements (also called food supplements or nutritional supplements) are preparations designed to supplement the diet and provide nutrients (such as vitamins, minerals, fiber, fatty acids or amino acids) that may be missing or may be inadequately consumed in a person's diet. Some countries define dietary supplements as foods, while others define them as medicines or natural health products. Supplements containing vitamins or dietary minerals are included as a class of food in the Codex Alimentarius, a collection of internationally recognized standards, codes of practice, guidelines and other recommended specifications involving food, food production and food safety. These manuscripts are prepared by the Codex Alimentarius Commission, which is funded by the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO).

[0146] Disease prevention or treatment

[0147] "Prevention" includes reducing the risk and / or severity of a disease or disorder. The terms "treatment", "relief" and "relief" include both preventive or prophylactic treatment (preventing and / or slowing the development of a target pathological condition or disorder) and curative, therapeutic or disease-modifying treatment, including curing, slowing down, alleviating the symptoms of a diagnosed pathological condition or disorder and / or interrupting the therapeutic measures of its progress; and including treating patients who are at risk of contracting a disease or suspected of contracting a disease, as well as treating patients who are sick or have been diagnosed with a disease or medical condition. The term does not necessarily mean that an individual is treated until full recovery. All of these terms also refer to the maintenance and / or promotion of health in individuals who do not have a disease but may be prone to unhealthy conditions. These terms are also intended to include strengthening or otherwise enhancing one or more major preventive or therapeutic measures. The terms "treatment", "relief" and "relief" are also intended to include dietary management of a disease or condition or dietary management that prevents or prevents a disease or condition. Treatment can be patient-related or doctor-related.

[0148] The present invention will now be further described by way of examples, which are intended to help those skilled in the art to practice the present invention but are not intended to limit the scope of the present invention in any way.

[0149] Example

[0150] Example 1

[0151] Metabolite measurements in serum

[0152] Twenty male participants, predominantly of Chinese descent, aged 65 to 79 years, were recruited from 2 studies of healthy older male community residents in Singapore (the Singapore Sarcopenia Group and the Aging in Community Environments Study [ACES]), and 20 control individuals of the same age group who were not diagnosed with sarcopenia were included. Sarcopenia was diagnosed first using the definition of the Asian Working Group on Sarcopenia in the Elderly (Chen et al., 2014) and then according to the standardized operational definition of sarcopenia (Cruz-Jentoft et al., 2010).

[0153] The National Health Care Group Domain-Specific Research Committee (NHG DSRB) approved the study (reference number 2014 / 01304), and each participant provided written informed consent. Weight and height were measured to the nearest 0.1 kg or 1 cm. Total lean body mass was measured by dual-energy X-ray absorptiometry (DXA) scanning (APEX software version 4.0.1, Discovery WiDXA system). Isometric grip strength was measured by Jamar handheld dynamometer using a standardized protocol, and the average of 3 attempts in the dominant hand was used as the final measurement. Physical fitness was assessed by measuring walking speed on a 6-meter habitual paced walk test. The diagnosis of sarcopenia was based on the AWGSOP definition, which is defined as total limb lean body mass normalized for height less than 7.00 kg / m2, evidenced by low physical fitness based on walking speed less than 0.8 m / sec, or low muscle strength based on grip strength less than 26 kg. The anthropometric parameters of the control and sarcopenia groups were as follows:

[0154]

[0155] Fasting blood samples were drawn from the antecubital fossa using a vacuum blood collection tube and an indwelling 20ga butterfly cannula. Blood was centrifuged at 3000rpm for 10 minutes at 4°C, and then serum was aliquoted and frozen at -80°C until further analysis. At BEVITAL Laboratory (Bergen, Norway), by liquid chromatography and then by mass spectrometry LC-MS / MS), the level of anthranilic acid and trigonelline in serum were measured. Semi-open muscle biopsies of the vastus lateralis of 20 male participants and 20 age-matched controls were collected using a BioPinceTM (Angiotech) 16G full-core biopsy needle with 3 adjustable stroke lengths (13mm, 23mm, 33mm), which were then snap-frozen in liquid nitrogen and stored at -80°C until further analysis.

[0156] After log2 transformation, the serum levels of trigonelline, anthranilic acid, and their ratio (serum level of trigonelline: serum level of anthranilic acid) were tested for differences between sarcopenia cases and controls using Wilcoxon test and Student's t test. We investigated the correlation of these analytes with a continuous clinical variable for characterizing sarcopenia (limb lean muscle mass index (ALMi; kg / m 2 ), grip strength, and walking speed).

[0157] Figures 1 to 3Serum levels of trigonelline, anthranilic acid, and the ratio of trigonelline:anthranilic acid differ in sarcopenic patients and are associated with lean muscle mass, grip strength, and gait. Trigonelline levels were measured in serum from a human population of 20 sarcopenic patients aged 65 years and older and 20 age-matched controls. Figure 1 ), the level of anthranilic acid ( Figure 2 ) and the ratio of trigonelline level / anthranilic acid level ( Figure 3 ). The left panel shows the log2 values ​​of the analytes in the serum of sarcopenic patients and control patients. The right panel shows the correlation of the analytes in serum with the limb lean mass index (ALMi), which was measured by DXA as a surrogate for muscle mass, grip strength and walking speed. The orange line shows the fitted regression line of the clinical variables of the analytes, and the gray area shows the 95% confidence interval of the fitted model. The reported P values ​​were calculated by Student's T statistic (case-control) or Pearson correlation (correlation analysis) on the Log2 transformed values.

[0158] Example 2

[0159] Muscle gene expression analysis

[0160] For muscle gene expression analysis, according to the manufacturer's instructions, total RNA was extracted from muscle biopsies using the Qiagen test kit. RNA quantity was measured with Ribogreen (Life Technologies), and RNA quality was checked using the standard sensitivity RNA analysis kit on a fragment analyzer (Advanced Analytical Technologies). All RNA samples were homogeneous and passed quality control, with 260 / 280nm ratios>1.8 and RIN scores>7. For RNA sequencing, 250ng total RNA from each sample was used as the raw material for library construction, and ribosomal RNA was removed using the Ribo-Zero magnetic test kit (Illumina). Sequencing libraries were constructed using the TruSeq Stranded RNA HT Gold Test Kit (Illumina) with Ribo-Zero, and 13 cycles of PCR amplification steps were then performed using KAPA HiFi HotStart ReadyMix (Kapa BioSystems). Libraries were quantified with Picogreen (Life Technologies) and size controlled with the DNA High Sensitivity Kit on LabChip GX (PerkinElmer). Libraries were then pooled in equimolar ratios and aggregated at 7 pmol on a double-end sequencing flow cell (Illumina). 2 × 101 cycles of sequencing were performed on a HiSeq 2500 (Illumina) equipped with V3 chemistry. The generated data were demultiplexed using Casava. Reads were aligned to the human genome (hs_GRCh38.p2) using STAR (Dobin et al., 2013), and the number of reads mapped within genes was quantified by HTSeq (Anders et al., 2015) (HTSeq-0.6.1p1 version, mode union, strand reverse, quality alignment greater than 10). The samples were sequenced to a depth of 75 to 104 million reads per sample, of which 34 to 77 million reads were uniquely mapped.

[0161] Differentially expressed genes between control and sarcopenic samples were defined by using the limma package (Smyth, 2004). Briefly, after removing genes with an average expression of less than 20 reads, the data were normalized by the M-value truncated mean (TMM) method as implemented in the edgeR function calcNormFactors (Robinson et al., 2010), and then the voomWithQualityWeights function was applied to model the mean-variance relationship and estimate sample-specific quality weights (Liu et al., 2015). P values ​​were corrected for multiple testing using the Benjamini-Hoechberg method. The same procedure was applied when the characterization of the association between gene expression and continuous or categorical parameters (ALMi, grip strength, and walking speed) was used to define sarcopenia. Gene set enrichment analysis was performed using CAMERA (Wu and Smyth, 2012), a competitive gene set test that queries whether a gene set annotated in the Molecular Signature Database (MSigDB) (Subramanan et al., 2005) is enriched for differentially expressed genes. Pathway analysis was performed using MSigDB version 5.2 collections H (featured gene set), C2 (curated gene set), and C5 (GO gene set).

[0162] In Figure 4, RNA sequencing of human sarcopenic skeletal muscle shows that mitochondrial dysfunction is the main transcriptional change during sarcopenia. (A) Gene set enrichment analysis was performed on sarcopenic muscle and control muscle using CAMERA and a signature gene set collection from MSigDB. The red bar represents the gene set enriched in sarcopenic muscle, and the blue bar represents the gene set suppressed in sarcopenic muscle. Gene sets are ranked according to the significance of their enrichment. (B) GSEA enrichment curves of the oxidative phosphorylation gene set selected from A.

[0163] Spearman rank correlations between TMM normalized muscle gene expression and each serum analyte of interest (trigonelline, anthranilic acid, and trigonelline: anthranilic acid ratio) were calculated and used to perform gene set enrichment analysis using the biosignature gene set collection.

[0164] Figure 4AThe keys: 1.OXIDATIVE_PHOSPHORYLATION,2.FATTY_ACID_METABOLISM,3.MYC_TARGETS_V1,4.ALLOGRAFT_REJECTION,5.MTORC1_SIGNALING,6.PEROXISOME,7.ADIPOGENESIS,8 .EPITHELIAL_MESENCHYMAL_TRANSITION,9.APICAL_JUNCTION,10.G2M_CHECKPOINT,11.BILE_ACID_METABOLISM,12.UNFOLDED_PROTEIN_RESPONSE,13.REACTIVE_OXIGEN_ SPECIES_PATHWAY,14.MITOTIC_SPINDLE,15.UV_RESPONSE_UP,16.UV_RESPONSE_DN,17.PI3K_AKT_MTOR_SIGNALING,18.MYC_TARGETS_V2,19.KRAS_SIGNALING_DN,20.HED GEHOG_SIGNALING,21.E2F_TARGETS,22.INTERFERON_GAMMA_RESPONSE,23.APICAL_SURFACE,24.TGF_BETA_SIGNALING,25.PROTEIN_SECRETION,26.IL2_STAT5_SIGNALING.

[0165] Fig. 5 to Fig. 7 show the correlation of skeletal muscle gene expression with the serum level of trigonelline, the serum level of anthranilic acid and the serum level of trigonelline: the ratio of the serum level of anthranilic acid.Gene set enrichment analysis (GSEA) of muscle gene expression related to the serum level of trigonelline (Fig. 5), the serum level of anthranilic acid (Fig. 6) and the ratio (Fig. 7) of the serum level of trigonelline / the serum level of anthranilic acid.GSEA is performed using the average rank gene set test and the biological feature gene set collection from MSigDB.Red bars represent the gene set whose expression in skeletal muscle is positively correlated with the serum level of analyte, and blue bars represent the gene set whose expression in skeletal muscle is negatively correlated with the serum level of analyte.Gene set is sorted according to its enrichment significance, and only the gene set of FDR<1% is shown.Left block represents the GSEA enrichment curve of oxidative phosphorylation gene set.

[0166] Figure 5AKey: 1. OXIDATIVE_PHOSPHORYLATION, 2. MTORC1_SIGNALING, 3. PROTEIN_SECRETION, 4. ADIPOGENESIS, 5. FATTY_ACID_METABOLISM, 6. MYC_TARGETS_V1, 7. PEROXISOME, 8. DNA_REPAIR, 9. GLYCOLYSIS, 10. MITOTIC_SPINDLE, 11. MYOGENESIS, 12. INTERFERON_GAMMA_RESPONSE, 13. PI3K_AKT_MTOR_SIGNALING, 14. WNT_BETA_CATENIN_SIGNALING, 15. INTERFERON_ALPHA_RESPONSE, 16. ALLOGRAFT_REJECTION, 17. BILE_ACID_METABOLISM, 18. ANDROGEN_RESPONSE, 19. CHOLESTEROL_HOMEOSTASIS

[0167] Fig. 6A Key: 1. OXIDATIVE_PHOSPHORYLATION, 2. FATTY_ACID_METABOLISM, 3. ADIPOGENESIS, 4. G2M_CHECKPOINT, 5. BILE_ACID_METABOLISM, 6. PEROXISOME, 7. MYOGENESIS

[0168] Fig. 7A Key: 1. OXIDATIVE_PHOSPHORYLATION, 2. FATTY_ACID_METABOLISM, 3. ADIPOGENESIS, 4. MTORC1_SIGNALING, 5. PEROXISOME, 6. PROTEIN_SECRETION, 7. MYOGENESIS, 8. G2M_CHECKPOINT, 9. MYC_TARGETS_V1, 10. BILE_ACID_METABOLISM, 11. DNA_REPAIR, 12. CHOLESTEROL_HOMEOSTASIS, 13. KRAS_SIGNALING_DN, 14. GLYCOLYSIS

[0169] Figure 8It was shown that human sarcopenic muscle has lower expression of genes such as NMAT1, NAMPT, and PNP, which control the biosynthesis of NAD from NAD precursors or from diet and NAD remediation. Gene expression in muscle was measured by high-coverage RNA sequencing. NMNAT: Nicotinamide nucleotide adenylyltransferase 1; NAMPT: Nicotinamide phosphoribosyltransferase; PNP: Purine nucleoside phosphorylase).

[0170] Example 3

[0171] Measurement of NAD+ Levels in Muscle Biopsies

[0172] To determine whether NAD levels could be an upstream trigger of mitochondrial markers in sarcopenic muscle, NAD was measured in muscle biopsy material from the elderly. NAD+ levels were measured in human muscle biopsies as described by Dall et al. in 2018. Briefly, 5 mg of muscle tissue from the remaining biopsy was lysed in 200 μL 0.6 M perchloric acid, and the supernatant was diluted 250-fold in 100 mM Na2HPO4 pH8.0. 100 μL of the diluted sample was combined with 100 μL of a reaction mixture (100 mM Na2HPO4 pH8, 2% ethanol, 90 U / mL alcohol dehydrogenase, 130 mU / mL diaphorase, 10 μM resazurin, 10 μM flavin mononucleotide, 10 mM nicotinamide), and the fluorescence increase (Ex 540 nm / Em 580) was measured over 10 minutes. NAD+ content was calculated by a standard curve and normalized to tissue weight.

[0173] Fig. 9 showed that NAD levels were significantly reduced (32%) in human skeletal muscle of elderly patients with sarcopenia. This large reduction in NAD+ levels in skeletal muscle of people with sarcopenia is the first to link low NAD+ levels to age-related pathologies in humans and is the first evidence of NAD deficiency in human skeletal muscle. Combined with the preclinical efficacy of NAD precursors such as NR and NMN on aging muscle in mice, this result will expand the clinical relevance of NAD biology in aging and guide the clinical translation of sarcopenia and age-related mobility deficits.

[0174] Example 4

[0175] Metabolite measurements in serum of different ethnic groups including women.

[0176] This example considered 70 participants, predominantly Caucasian, 52 females and 18 males, with a median age of 78 years (interquartile range 76, 81).

[0177] The 70 participants were part of the Hertfordshire Sarcopenia Study (HSS), a substudy of the UK Hertfordshire Cohort Study (HCS) investigating the effects of life stage on muscle morphology, mass and strength in older community-dwelling adults, or were part of the HSSe extension phase of the study. Skeletal muscle characterization performed in this study included body composition and lean mass determined by dual-energy X-ray absorptiometry (DXA) scanning.

[0178] Fasting blood samples were drawn from the antecubital fossa using a vacuum blood collection tube and an indwelling 20ga butterfly cannula. The blood was centrifuged at 3000 rpm for 10 minutes at 4°C, and the serum was then aliquoted and frozen at -80°C until further analysis. Anthranilic acid levels and trigonelline levels in serum were measured by liquid chromatography followed by mass spectrometry (LC-MS / MS) at BEVITAL Laboratories (Bergen, Norway).

[0179] The relationship between trigonelline:anthranilic acid ratio and limb lean muscle mass index (ALMi; kg / m 2 ) were estimated as follows: after performing a log2 transformation on the serum concentrations of the analytes, regression adjusted for age was applied.

[0180] Fig.10 It was shown that the ratio of serum levels of trigonelline:anthranilic acid is associated with lean muscle mass estimated by the limb lean mass index. The levels of trigonelline and anthranilic acid in the serum of a human population of 70 individuals were measured, of which 52 were women and 18 were men, all aged 65 years and over. The graph represents the correlation of the log2 value of the ratio of the analytes in serum (trigonelline:anthranilic acid) (x-axis) with the limb lean mass index (ALMi) (y-axis), measured by DXA as a surrogate for muscle mass. The line shows the fitted regression line of ALMi for log2(trigonelline / anthranilic acid). From a simple linear regression, we obtained a Pearson correlation coefficient of 0.23 with a p-value = 0.031; based on the regression adjusted for age, we obtained an adjusted R of 51%. 2 , and the p-value for the coefficient relative to the analyte ratio = 0.024.

[0181] References

[0182] Anders, S., Pyl, PT, and Huber, W. (2015). HTSeq--a Python framework to work with high-throughput sequencing data. Bioinformatics 31, 166-169.

[0183] Anker, SD, Morley, JE, & von Haehling, S. (2016). Welcome to the ICD-10 code for sarcopenia. Journal of cachexia, sarcopenia and muscle 7, 512-514.

[0184] Chen, LK, Liu, LK, Woo, J., Assantachai, P., Auyeung, TW, Bahyah, KS, Chou, MY, Chen, LY, Hsu, PS, Krait, O., et al. (2014). Sarcopenia in Asia: consensus report of the Asian Working Group for Sarcopenia. Journal of the American Medical Directors Association 15, 95-101.

[0185] Clark RV, Walker AC, O'Connor-Semmes RL, Leonard MS, Miller RR, Stimpson SA, Turner SM, Ravussin E, Cefalu WT, Hellerstein MK, Evans WJ, Total body skeletal muscle mass: estimation by creatine (methyl-d3) dilution in humans. J Appl Physiol (1985). 2014 Jun 15; 116 (12): 1605-13.

[0186] Cruz-Jentoft, AJ, Baeyens, JP, Bauer, JM, Boirie, Y., Cederholm, T., Landi, F., Martin, FC, Michel, JP, Rolland, Y., Schneider, SM, et al. (2010). Sarcopenia: European consensus on definition and diagnosis: Report of the European Working Group on Sarcopenia in Older People. Age Ageing 39, 412-423.

[0187] Dall, M., Penke, M., Sulek, K., Matz-Soja, M., Holst, B., Garten, A., Kiess, W., and Treebak, J.T. Hepatic NAD(+) levels and NAMPT abundance are unaffected during prolonged high-fat diet consumption in C57BL / 6JBomTac mice. Molecular and Cellular Endocrinology (2018) S0303-7207(18)30048.

[0188] Dobin, A., Davis, C. A., Schlesinger, F., Drenkow, J., Zaleski, C., Jha, S., Batut, P., Chaisson, M., and Gingeras, T. R. (2013). STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29, 15-21.

[0189] Liu, R., Holik, AZ, Su, S., Jansz, N., Chen, K., Leong, HS, Blewitt, ME, Asselin-Labat, ML, Smyth, GK, and Ritchie, ME (2015). Why weight? Modelling sample and observational level variability improves power in RNA-seq analyses. Nucleic acids research 43, e97.

[0190] Robinson, MD, McCarthy, DJ, and Smyth, GK (2010). edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139-140.

[0191] Smyth, GK (2004), Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Statistical applications in genetics and molecular biology 3, Article 3 (Linear models and empirical bayes methods for assessing differential expression in microarray experiments, Statistical applications in genetics and molecular biology, volume 3, article 3).

[0192] Studenski SA, Peters KW, Alley DE, Cawthon PM, McLean RR, Harris TB, Ferrucci L, Guralnik JM, Fragala MS, Kenny AM, Kiel DP, Kritchevsky SB, Shardell MD, Dam TT, Vassileva MT (2014). The FNIH sarcopenia project: rationale, study description, conference recommendations, and final estimates. J Gerontol A Biol Sci Med Sci. 69(5), 547-558.

[0193] Stimpson SA, Leonard MS, Clifton LG, Poole JC, Turner SM, Shearer TW, Remlinger KS, Clark RV, Hellerstein MK, Evans WJ. (2013) Longitudinal changes intotal body creatine pool size and skeletal muscle mass using the D3-creatine dilution method. J Cachexia Sarcopenia Muscle. Jun 25.

[0194] Subramanian, A., Tamayo, P., Mootha, VK, Mukherjee, S., Ebert, BL, Gillette, MA, Paulovich, A., Pomeroy, SL, Golub, TR, Lander, ES, et al. (2005). Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proceedings of the National Academy of Sciences of the United States of America 102, 15545-15550.

[0195] Wu, D. and Smyth, GK (2012). Camera: a competitive gene set test accounting for inter-gene correlation. Nucleic acids research 40, e133.

Claims

1. Use of trigonelline as a biomarker in the preparation of a product for determining whether an individual suffers from sarcopenia or has an increased risk of developing sarcopenia, comprising: (a) determining the level of trigonelline in a sample obtained from the individual; (b) comparing the level of trigonelline in the sample with a reference value; Wherein a reduced level of trigonelline in the sample compared to the reference value is an indication that the individual suffers from sarcopenia or is at risk of developing sarcopenia.

2. The use according to claim 1, comprising: (c) further determining the level of anthranilic acid in a sample obtained from the individual; (d) determining the ratio of trigonelline to anthranilic acid in the sample; (e) comparing the ratio of trigonelline to anthranilic acid in the sample with a reference value; wherein the ratio of trigonelline to anthranilic acid in the sample compared to the reference value is an indication that the individual suffers from or is at risk of developing sarcopenia.

3. The use according to claim 2, wherein a decrease in the ratio of trigonelline to anthranilic acid in a sample from the individual compared to the reference value is an indication that the individual suffers from sarcopenia or has an increased risk of developing sarcopenia.

4. The use according to any one of claims 1 to 3, wherein sarcopenia is characterized by one or more of low muscle mass, low muscle strength and low physical fitness.

5. The use according to claim 4, wherein (i) muscle mass is measured by DXA, MRI or D3 creatine dilution; (ii) muscle strength is measured by grip strength or knee extensor strength; and (iii) physical fitness is measured by walking speed, SPPB, 400 m walk test, timed up and go test or stair climbing test.

6. Use according to any one of claims 1 to 3, wherein the level of trigonelline and / or the level of anthranilic acid is determined by mass spectrometry.

7. The use according to any one of claims 1 to 3, wherein the individual is a human individual.

8. The use according to claim 7, wherein the human subject is a middle-aged or elderly person.

9. The use according to any one of claims 1 to 3, wherein the reference value is determined from a sample obtained from the same individual or a group of individuals.

10. The use according to any one of claims 1 to 3, wherein the sample obtained from the individual is a blood-derived sample.

11. The use according to any one of claims 1 to 3, wherein the sample obtained from the individual is a urine sample.

12. Use of trigonelline as a biomarker in the preparation of a product for predicting the response of an individual suffering from sarcopenia or at increased risk of developing sarcopenia to a nutritional composition for treating or preventing sarcopenia, comprising: (a) detecting the level of trigonelline in a sample obtained from the individual; (b) comparing the level of trigonelline in the sample with a reference value; (c) predicting a response of the individual to the nutritional composition based on the results of step (b); Wherein if the level of trigonelline in the sample is reduced compared to the reference value, it will be predicted that the individual will respond to the nutritional composition.

13. Use of trigonelline and anthranilic acid as biomarkers in the preparation of a product for predicting the response of an individual suffering from sarcopenia or at increased risk of developing sarcopenia to a nutritional composition for treating or preventing sarcopenia, comprising: (a) detecting the levels of trigonelline and anthranilic acid in a sample obtained from the individual; (b) comparing the ratio of trigonelline to anthranilic acid in the sample with a reference value; (c) predicting a response of the individual to the nutritional composition based on the results of step (b); Wherein if the ratio of trigonelline to anthranilic acid in the sample is reduced compared to the reference value ratio, it will be predicted that the individual is responsive to the nutritional composition.

14. Use according to claim 12 or 13, wherein the nutritional composition comprises vitamin B3.

15. Use according to claim 12 or 13, wherein the nutritional composition comprises a NAD precursor.

16. The use according to claim 15, wherein the NAD precursor is nicotinamide riboside.

17. The use according to claim 12, wherein the nutritional composition comprises trigonelline.

18. The use according to claim 12, wherein the nutritional composition comprises coffee extract.

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