Nutrional composition for treating and / or preventing muscle wasting

Abstract

The invention pertains to a nutritional composition comprising taurine, carnitine and a mixture of low- molecular-weight polyphenols derived from lychee fruit and green tea leaves. Further, the invention pertains to a nutritional composition comprising taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves for use as a medicament, more particularly for therapeutic use in treating and / or preventing muscle wasting.

Description

NUTRIONAL COMPOSITION FOR TREATING AND / OR PREVENTING MUSCLE WASTINGField of the inventionThe invention relates to a nutritional composition for prevention and / or treatment of muscle wasting or muscle decline, particularly for therapeutic use in improving muscle mass, strength and function.Background of the inventionSufficient muscle energy production and muscle mass is critical to the ageing population. Muscle mass loss becomes pathologic during oncology, hospitalized immobile patients, patients suffering from inflammatory diseases, malnourished elderly or patients suffering from immobilizing diseases like osteoarthritis or rheumatoid arthritis.. Muscle functionality and physical ability decline as a result of impaired muscle energy production and muscle mass loss. Muscle functionality and physical ability decline as a result of impaired muscle energy production and muscle mass.Sarcopenia is a progressive and generalized skeletal muscle disorder characterized by the accelerated loss of muscle mass, strength, and function. It primarily affects older adults and is associated with aging. This condition can significantly impact quality of life by reducing physical capabilities and increasing the risk of falls and fractures. Frailty and sarcopenia are challenges that come with age, and accelerated by acute or chronic diseases. From the age of about 30, humans lose 3-8 % of muscle per decade and this accelerates after 60 years of age.Muscle wasting and weakness is an important medical problem of critically ill patients. Intensive care unit (ICU) acquired weakness, the clinical diagnosis of muscle weakness, occurs in critically ill patients. The ICU patient population is very diverse where different events (e.g. trauma, scheduled operation) can attribute to an admission of a patient to the ICU. The first days in the ICU are considered as acute phase. During the acute phase a lot is happening which can challenge (nutritional) interventions (i.e. inflammatory storm, anxiety, organ failure). Therefore, muscle wasting, high levels of inflammation and oxidative damage are observed in ICU patients during post ICU recovery when the patients are stable.Energy supply, hence mitochondrial function is crucial to skeletal muscle health and detrimental changes at the level of the mitochondria may contribute to loss of muscle mass and functionality. A main function of mitochondria is the generation of ATP for energy through oxidative phosphorylation (OXPHOS). Mitochondrial dysfunction is associated with impaired immune and inflammatory responses. Chronic inflammation due to the enhanced production of inflammatory cytokines causes muscle wasting with a reduction in muscle mass and power. Muscle wasting caused by inflammatory cytokines is defined as cytokine-induced muscle wasting. The changes of mitochondrial network influence the production of reactive oxygen species (ROS) that play an important role in muscle function. It is shown that mitochondrial bioenergetics are altered in aged skeletal muscle, resulting in an increase in ROS production, while conversely genetic / pharmacological approaches that attenuate mitochondrial ROS promote healthy aging and maintenance of muscle mass. Also, exercise is asuccessful strategy to improve mitochondrial function by activating mitochondrial biogenesis and mitophagy. However, exercise is not always an option for the (post) ICU patient. Caloric restriction has also shown a beneficial effect on mitochondrial function but this is not a recommended strategy for already malnourished (post) ICU patient.There is a need for a nutritional composition which can support muscle function by stimulating the mitochondria for treating and / or preventing muscle wasting in the ageing population or oncology or hospitalized patients.Muscle function, sometimes in the context of sarcopenia, is addressed by Alessandra etal. in "Taurine Atenuates Catabolic Processes Related to the Onset of Sarcopenia" Int. J. Mol. Sci., vol. 21 (22) 2020 page 8865, and in "Taurine Administration Counteracts Aging-Associated Impingement of Skeletal Muscle Regeneration by Reducing Inflammation and Oxidative Stress" Antioxidants vol.11 (5) 2022 page 1016; by Yoshiharu et al. "Association of carnitine insufficiency with sarcopenia and dynapenia in patients with heart failure" Geriatrics and Gerontology Int. vol 23(7) 2023, pages 524-530; by Ning et al "Disturbed carnitine metabolism is independently correlated with sarcopenia and prognosis in patients on hemodialysis" Clin. Nutr. Vol. 43(9) 2024, pages 2019-2027; by Taotao etal. "Mechanisms Underlying the Effects of the Green Tea Polyphenol EGCG in Sarcopenia Prevention and Management" J. Agricultural and Food Chemistry vol. 71(25) 2023, pages 9609-9627; by Medoro etal. "Polyphenol Supplementation and Sarcopenia: A Systematic Review and Meta-Analysis of Clinical Trials" J. Frailty & Ageingvol. 13(4) 2024, pages 432-440; by Yun-Ching etal. "Oligonol Alleviates Sarcopenia by Regulation of Signaling Pathways Involved in Protein Turnover and Mitochondrial Quality” Mol. Nutr. & Food Research vol. 63(10) 2019; and by Sara et al. "Polyphenols and their potential role in preventing skeletal muscle atrophy" Nutrition Research Vol 74 (2019), pages 10-22.Outside the field, DE20309608 relates to compositions comprising soy protein, omega-3 fatty acid, arginine and / or carnitine; cacao is one of many additional ingredients that can be added, mentioned as a source of polyphenols such as proanthocyanidin oligomers (OPC).Summary of the inventionThe present invention provides a nutritional composition comprising taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves, also characterized as a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer, commercialized as Oligonol®. The inventors have found that the combination of taurine, carnitine and a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer improves mitochondrial function in an inflammatory context and hence, helps for treating and / or preventing muscle wasting. Reference is made to the example and Figure 1 herein. While all components have an effect beyond the reference,the combination has a significant, synergistic effect that goes beyond an expected additive effect. The model used to test synergy is an Agilent Seahorse Cell Mito Stress Test kit which is a widely recognized, well accepted standard assay for assessing mitochondrial function. The assay provided insights into the mechanism of mitochondrial dysfunction.Therefore, the present invention provides also a nutritional composition comprising taurine, carnitine and the mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer and / or derived from lychee fruit and green tea leaves for use as a medicament. The invention further provides a nutritional composition comprising the aforementioned combination for use in the treatment and / or prevention of muscle wasting in a human subject. Taurine, carnitine and the aforementioned mixture are present in therapeutically effective amounts.List of figuresThe Figure (Figure 1) shows the effect of taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves (oligonol) on mitochondrial function in cytokine-induced muscle wasting. For convenience sake, the synergistic effect beyond additive effect expected based on the individual contributions of the three components is marked in the figure: reference (vehicle) 1.88, muscle wasting (trigger) 0.99, treatment with taurine 1.13, carnitine 1.18, oligonol 1.09 and the combination 1.52. The effect could not be attributed to EGCG which is an important component in green tea. Green tea in combination with carnitine and taurine did not trigger any effect beyond that of the individual components (not shown).List of preferred embodiments1. A nutritional composition comprising taurine, carnitine and a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer.2. A nutritional composition according to embodiment 1 , wherein the mixture of low-molecular-weight polyphenols comprises 13-18 % flavanol monomers, 14-18 % dimers and 2-6 % trimers.3. A nutritional composition according to embodiment 1 or 2, wherein the mixture of low-molecular-weight polyphenols is obtainable by heating lychee fruit with green tea leaves in an acidic aqueous solution.4. The nutritional composition according to any one of the preceding embodiments wherein the composition comprises a protein and preferably the protein is selected from whey protein, soy protein, pea protein, canola protein or mixtures thereof.5. The nutritional composition according to embodiment 4 wherein the composition comprises at least 10 g protein per 100 g dry weight, preferably at least 15 g per 100 g dry weight.6. The nutritional composition according to any one of the preceding embodiments wherein the composition further comprises a coenzyme Q10.7. The nutritional composition according to any one of the preceding embodiments wherein the composition comprises, per 100 ml or per 150 kcal, 50 to 300 mg taurine, 50 to 600 mg carnitine and 5 to 30 mg of a mixture of low-molecular-weight polyphenols, preferably 50 to 300 mg taurine, 100 to 600 mg carnitine and 5 to 30 mg of a mixture of low-molecular-weight polyphenols.8. The nutritional composition according to any one of the preceding embodiments, wherein the composition is an oral nutritional supplement or a tube feeding.9. The nutritional composition according to any one of the preceding embodiments, for use as a medicament.10. The nutritional composition according to embodiments 1 to 9, for use in the treatment and / or prevention of muscle wasting in a human subject.11. The nutritional composition for use according to embodiment 10, wherein the human subject is an elderly subject of at least 50 years of age, preferably at least 55 years of age; and / or wherein the human subject is suffering from muscle decline; and / or wherein the human subject is suffering from cancer or sarcopenia or Intensive Care Unit (ICU) acquired weakness.12. Use of a nutritional composition comprising therapeutically effective amounts of taurine, carnitine and a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer, preferably derived from lychee fruit and green tea leaves, in the manufacture of a product for prevention and / or treatment of muscle wasting in a human subject.13. A method of prevention and / or treatment of muscle wasting in a human subject comprises administering a nutritional composition comprising therapeutically effective amounts of taurine, carnitine and a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer, preferably derived from lychee fruit and green tea leaves, to the human subject.14. The nutritional composition according to any one of embodiments 1 - 8, wherein the composition comprises a protein fraction comprising 32-58 weight % pea protein, 30-56 weight % soy protein and 8-16 weight % free leucine, preferably a protein fraction comprising 36-54 weight % pea protein, 34-52 weight % soy protein and 10- 14 weight % free leucine, and optionally upto 5 weight % of free branched chain amino acids other than leucine, based on all proteinaceous matter, wherein the composition is essentially free from animal protein; wherein the composition preferably comprises a protein fraction comprising 41-50 weight % pea protein, 39-47 weight % soy protein and 10.5 - 13.5 weight % free leucine, based on all proteinaceous matter; wherein the composition preferably comprises a protein fraction comprising 43-47 weight % pea protein, 41-45 weight % soy protein and 11 - 13 weight % free leucine, based on all proteinaceous matter.15. The nutritional composition according to any one of embodiments 1 - 8, wherein the composition comprises a protein fraction comprising (a) 15 to 35 weight % of casein ; (b) 25 to 50 weight % of whey protein ; (c) 10 to 30 weight % of soy protein ; and (d) 10 to 30 weight % of pea protein, relative to the total protein in the protein fraction, wherein the sum of said proteins preferably equals 100 weight % of the protein fraction; wherein the composition preferably comprises a protein fraction comprising (a) 20 to 30 weight % of casein ; (b) 30 to 40 weight % of whey protein ; (c) 15 to 25 weight % of soy protein ; and (d) 15 to 25 weight % of pea protein, relative to the total protein in the protein fraction, wherein the sum of said proteins preferably equals 100 weight % of the protein fraction.Detailed description of the inventionThe invention relates to a nutritional composition comprising therapeutically effective amounts taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves. The inventors of the present invention have found that a nutritional composition comprising taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves, improves mitochondrial function and hence, helps for treating and / or preventing muscle wasting in a human subject in need thereof. The (prophylactic) treatment of muscle wasting includes therapeutic use in improving muscle mass, strength and function.In an embodiment, the nutritional composition according to the invention is for use as a medicament. In a preferred embodiment, the nutritional composition is for use in the treatment and / or prevention of muscle wasting in a human subject.The subject is preferably an elderly human. In this respect, it is submitted that in the context of this application, an elderly human is a person of the age of 50 years or more, in particular of the age of 55 or more, more in particular of the age of 60 or more, more in particular of the age of 65 or more. This definition takes into account the fact that the average age varies between different populations, on different continents, etc. Most developed world countries have accepted the chronological age of 65 years as a definition of 'elderly' or older person (associated with the age at which one may begin to receive pension benefits), but like many westernized concepts, this does not adapt well to e.g. the situation in Africa. At the moment, there is no United Nations (UN) standard numerical criterion, but the UN agreed cut-off is 60+ years to refer to the older population in Western world. The more traditional African definitions of an elder or 'elderly' person correlate with the chronological ages of 50 to 65 years, depending on the setting, the region and the country.In a preferred embodiment said human subject is suffering from muscle decline. In a further preferred embodiment, the human subject is suffering from cancer or sarcopenia or ICU acquired weakness. Sarcopenia, the loss of muscle mass and strength, becomes more common with age. Pathologic muscle mass loss in sarcopenia is typically defined by a reduction in muscle mass that is more than two standard deviations below the mean for young, healthy adults. This significant loss of muscle mass is often accompanied by a decrease in muscle strength and physical performance, which can severely impact daily activities and overall quality of life. Excessive muscle loss is commonly observed in cancer patients. The mechanisms of cancer-related muscle loss are multifactorial, including anorexia, hypogonadism, anaemia, inflammation, malnutrition, and aberrations in skeletal muscle protein turnover and metabolism. Also, low muscle in cancer is associated with an increase in treatment-related toxicities. ICU acquired weakness, the clinical diagnosis of muscle weakness, is observed in approximately half of the ICU patients. Muscle mass decreases rapidly after ICU admission.CompositionThe nutritional composition according to the invention comprises taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves. These are present in therapeutically effective amounts, meaning that the components are presented in amounts that they altogether result in a reducing (the risk of) muscle wasting beyond the results achieved for corresponding amounts of the individual components.The first component, taurine, 2-aminoethanesulfonic acid, is a naturally occurring sulphur-containing amino acid that helps with bodily functions, including those of the digestive, cardiovascular, skeletal, muscular and nervous system. Taurine also has an antioxidant role and potentially suppresses excessive oxidants generation in the mitochondria and maintains the functionality of mitochondria. Although taurine is abundant in human organs with diverse putative roles, it is not an essential human dietary nutrient and is not included among nutrients with a recommended intake levels.The second component, carnitine is a quaternary ammonium compound involved in metabolism in most mammals, plants and some bacteria. In support of energy metabolism, carnitine transports long-chain fatty acids from the cytosol into mitochondria to be oxidized for free energy production, and also participates in removing products of metabolism from cells. Carnitine is concentrated in tissues like skeletal and cardiac muscle and is known for its role in helping to transport long-chain fatty acids into mitochondria for energy production. Therefore, carnitine is often associated with improved endurance and reduced muscle fatigue.In one embodiment, the third component is a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves. In the experiment, a commercially available mixture Oligonol® is used. Oligonol is a mixture of low-molecular-weight polyphenols derived from lychee fruit (Litchi chinensis) and green tea leaves (Camellia sinensis). Oligonol can be obtained by an oligomerization reaction which cleaves the polyphenols present in a combined extract of lychee fruit and green tea leaves into monomers and low molecular weight oligomers. Oligonol has 2 to 8 times higher proportion of monomers and oligomers than the other polyphenols. The phenolic mixture of oligonol comprises 13-18 % flavanol monomers, 14-18 % dimers and 2-6 % trimers. By contrast, unprocessed lychee fruit extract contains only 6.4 % monomers and 9.9 % dimers. Therefore, the bioavailability of oligonol is higher than unprocessed lychee fruit polyphenols.Among polyphenols, proanthocyanidins are naturally occurring plant metabolites and widely available in fruits, vegetables, nuts, seeds and flowers. In order for proanthocyanidins to exhibit various physiological activities, proanthocyanidin compound needs to be absorbed into the living body by the gastrointestinal tract. However, molecular weights of proanthocyanidins are generally said to be on the order of several thousands to several tens of thousands. Substance having such a large molecular weight is difficult to be absorbed by the gastrointestinal tract and in many cases, even if it is ingested, it is not absorbed in the living body and not used as nutrition.In one embodiment, the third component is a mixture of low-molecular-weight polyphenols as is described in EP1852430A - its whole contents herein incorporated by reference - and comprises proanthocyanidin oligomer to which a substance having a phloroglucinol ring structure or resorcinol ring structure has been bonded. This mixture comprising the proanthocyanidin oligomer is preferably obtained or obtainable by heating, for example at 90 to 100 degrees centigrade for 1 to 4 hours, plant materials comprising proanthocyanidin polymer or extract thereof comprising proanthocyanidin polymer, with a substance having a phloroglucinol ring structure or resorcinol ring structure in an acidic aqueous solution. The resulting mixture may then be filtered, concentrated, fractionated or dried using standard techniques known to the skilled person. Such a composition has been reduced in molecular weight compared to the proanthocyanidin polymer feed stock to such a level that the oligomer can be more easily absorbed into a living body. The plant containing proanthocyanidin polymer is preferably grape, pine, Chamaecyparis obtuse, camphor tree, wax myrtle, cacao, date plum, banana, Chinese quince, apple, hawthorn, litchee, Myrica rubra and / or Cinnamomi Cortex, and most preferably derived from lychee fruit and green tea leaves, as is the commercially available Oligonol® mixture.In a preferred embodiment, a composition containing a mixture of low-molecular-weight polyphenols is obtained or obtainable by heating lychee fruit with green tea leaves in an acidic aqueous solution (pH < 6). The heating can be done around 90 to 100 degrees centigrade for 1 to 4 hours. Acidic aqueous solution can be obtained by using an acid selected from inorganic acids such as hydrochloric acid, sulfuric acid and nitric acid and organic acids such as acetic acid, citric acid, ascorbic acid and malic acid.Alternatively, the third component may be described as a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer represented by the formula (1) or (2):and / or5 wherein n in formula (1) or (2) is an integer and is 0, 1 or 2.A best mode for preparing the third component is disclosed in paragraphs

[0015] to

[0023] of EP1852430A1 and is incorporated herein by reference. In order to obtain oligonol, lychee fruit can be used as a plant material comprising proanthocyanidin polymer and green tea leaves can be used as a substance having a phloroglucinol ring structure or resorcinol ring structure. The proportion of proanthocyanidin polymer-containing plant materials and substance having phloroglucinol ring structure or resorcinol ring structure used in the reaction is arbitrarily selected. It is preferred that the amount of the latter be large enough to bond to the fragments of the proanthocyanidin polymer having reduced in the molecular weight. If the amount of the substance having phloroglucinol ring structure or resorcinol ring structure is too small, proanthocyanidin having a high molecular weight may remain unreacted, and in that case, the remaining proanthocyanidin having a high molecular weight can be easily removed by column chromatography. While arbitrarily selected, the proportion of lychee fruit and green tea leaves used to obtain the mixture of low-molecular-weight polyphenols is preferably between 1 :10 and 10:1 , more preferably between 1 :1 and 10:1. In EP1852430A1 , a ratio of 5:1 is applied. The skilled person can, without inventive merit, determine the appropriate ratio of lychee fruit and green tea leaves to produce the mixture comprising the proanthocyanidin oligomer according to the invention.In the above, the terminology ‘mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves’ and ‘mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer’ are used interchangeable and reflect the third component (Oligonol) in the composition of the invention. Preferably, the mixture comprises 13-18 % flavanol monomers, 14-18 % dimers and 2-6 % trimers.In one embodiment, the nutritional composition according to the invention comprising taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves or mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer as described here above.In one embodiment, the nutritional composition according to the invention comprising 50 to 300 mg / 100 mL taurine, 50 to 600 mg / 100 mL carnitine and 5 to 30 mg / 100 mL of the mixture of low-molecular-weight polyphenols as described herein, preferably 100 to 200 mg / 100 mL taurine, 100 to 400 mg / 100 mL carnitine and 5 to 20 mg / 100 mL of the mixture of low-molecular-weight polyphenols as described herein. More preferably, the nutritional composition according to the invention comprising 120 to 180 mg / 100 mL taurine, 120 to 180 mg / 100 mL carnitine and 10 to 20 mg / 100 mL of the mixture of low-molecular-weight polyphenols as described herein.Alternatively, the nutritional composition according to the invention comprises, per 150 kcal, 50 to 300 mg taurine, 50 to 600 mg carnitine and 5 to 30 mg of the mixture of low-molecular-weight polyphenols as described herein, preferably 50 to 300 mg taurine, 100 to 600 mg carnitine and 5 to 30 mg of the mixture of low-molecular-weight polyphenols, more preferably 100 to 200 mg taurine, 100 to 400 mgcarnitine and 10 to 20 mg of the mixture of low-molecular-weight polyphenols as described herein. Most preferably, the nutritional composition according to the invention comprises, per 150 kcal, 120 to 180 mg taurine, 120 to 180 mg carnitine and 10 to 20 mg of the mixture of low-molecular-weight polyphenols as described herein.Alternatively, the nutritional composition according to the invention comprises, per unit dose, 100 to 400 mg taurine, 100 to 400 mg carnitine and 5 to 40 mg of the mixture of low-molecular-weight polyphenols as described herein. Preferably, the nutritional composition according to the invention comprises, per unit dose, 120 to 360 mg taurine, 120 to 360 mg carnitine and 10 to 40 mg of the mixture of low-molecular-weight polyphenols as described herein. The expression "unit dose" refers to an amount of a composition administered to an individual in a single dose. A daily dose is the sum of all single doses in a day. The unit dose is preferably in liquid form, with a volume between 100 and 200 ml.For all the individual nutrients a dose response experiment was initially performed on muscle cell viability. This was done to exclude any cytotoxicity of the single nutrient on the muscle cells. Once it was established that a specific dose or range of dose was safe, testing was continued in the Seahorse Mito Stress kit assay, subsequently multiple combinations were tested. A synergistic effect was observed for the mixture. Taking from these results and what has been used before in clinical studies for the single nutrients of interest, and a metanalysis on the benefits of taurine for metabolic syndrome and recommended supplementation in case of carnitine deficiency, the inventors established the therapeutic doses as described here above.It is preferred that the composition for use or method comprises administration of 100 - 4000 mg taurine per day, 100 - 4000 mg carnitine per day and 10-500 mg for the mixture of low-molecular-weight polyphenols (oligonol) per day, more preferably 100 - 2000 mg taurine per day, 100 - 2000 mg carnitine per day and 20-500 mg for the mixture of low-molecular-weight polyphenols (oligonol) per day.Alternatively or additionally, the nutritional composition according to the invention comprises taurine, carnitine and the mixture of low-molecular-weight polyphenols as described herein in a weight ratio of 1 : 10 - 100 : 0.01 - 0.1 , more preferably 1 : 10 - 20 : 0.05 - 0.1. The above limitations per 100 ml, 150 kcal and per day could be limited according to those ratios.According to a further embodiment, the nutritional composition according to the invention comprises protein selected from whey protein, soy protein, pea protein, canola protein or mixtures thereof.Whey protein is known for its excellent amino acid profile, and for its ability to increase the protein synthesis in a mammal (due to a higher leucine content). Nutritionally speaking, whey protein is known as a naturally complete protein because it contains all of the essential amino acids required in the daily diet. It is also one of the richest sources of branched chain amino acids (BCAAs, in particular leucine)which play an important role in muscle protein synthesis. Whey protein is the preferred choice of proteins to treat persons suffering from sarcopenia.Soy protein is categorized as a high-quality, complete protein although the methionine level is slightly below the WHO 2007 recommendation for methionine content. Soy proteins can be divided into different categories according to their production method. Soy protein isolate (SPI) is the most refined form of soy protein. Soy protein isolate contains about 90 % protein. Soy protein concentrate (SPC) is basically soybean without the water soluble carbohydrates. It contains about 70 % of protein.Pea protein contributes to forming a good overall mix of amino acids. While whey proteins enter the blood stream very fast, pea proteins are absorbed much slower. Pea protein is quite high in cysteine content and can therefore compensate the inadequate amount of cysteine in casein proteins. Furthermore, pea protein is quite high in arginine compared to casein, soy or whey protein which is required for muscle metabolism and which facilitates the intake of body mass while reducing body fat; and it is quite high in lysine, which is needed to build protein muscle and assist in the maintenance of lean body mass.Canola protein is rich in essential amino acids, making it a complete protein. The average protein content of canola is 20 %. The nutritional quality of canola protein, in terms of Protein Digestibility Corrected Amino Acid Score (PDCAAS), is comparable to that of soy protein. As a complete protein, it provides all the essential amino acids necessary for muscle repair and growth.In one embodiment, the nutritional composition comprises a protein fraction comprising (a) 15 to 35 weight % of casein ; (b) 25 to 50 weight % of whey protein ; (c) 10 to 30 weight % of soy protein ; and (d) 10 to 30 weight % of pea protein ; relative to the total protein in the protein fraction, wherein the sum of said proteins preferably equals 100 weight % of the protein fraction. The protein fraction preferably comprises (a) 20 to 30 weight % of casein ; (b) 30 to 40 weight % of whey protein ; (c) 15 to 25 weight % of soy protein ; and (d) 15 to 25 weight % of pea protein ; relative to the total protein in the protein fraction, wherein the sum of said proteins preferably equals 100 weight % of the protein fraction. According to a preferred embodiment, the proteins including casein, whey protein, soy protein and pea protein are substantially in intact form or non-hydrolysed. In the context of the invention, a "non-hydrolysed" protein is equivalent to an "intact" protein, meaning that the protein has not been subjected to an hydrolysis process. However, minor amounts of hydrolysed proteins may be present in the source of non-hydrolysed proteins, or may be added to the formulation, such as additional amino acids, such as, for example leucine, isoleucine, glutamine, arginine, or dipeptides and the like. In one embodiment of the present invention, the composition may comprise a free amino acid, or a mixture of free amino acids, up to about 1 g / 100 ml, preferably up to about 0.5 gram / 100 ml. The protein fraction preferably essentially consists of vegetable and dairy proteinaceous matter, in particular proteins, preferably casein, whey protein, soy protein and pea protein. Small amounts of free amino acids may be accounted for, preferably less than about 5 %, more preferably less than about 2 %, most preferablyless than about 1 % of the protein fraction. In a particularly preferred embodiment, there are no free amino acids, peptides or hydrolysates in the composition. Within the context of this paragraph, with the term "casein" both caseinate and micellar casein is indicated. In one embodiment, the casein is caseinate, preferably Na- caseinate or Ca-caseinate. Preferably, the caseinate is Ca-caseinate.In another embodiment, the nutritional composition comprises a protein fraction comprising 32-58 weight % pea protein, 30-56 weight % soy protein and 8-16 weight % free leucine, preferably a protein fraction comprising 36-54 weight % pea protein, 34-52 weight % soy protein and 10 - 14 weight % free leucine, and optionally up to 5 weight % of free branched chain amino acids other than leucine, based on all proteinaceous matter, wherein the composition is essentially free from animal protein. In a preferred embodiment, the composition comprises a protein fraction comprising 41-50 weight % pea protein, 39-47 weight % soy protein and 10.5 - 13.5 weight % free leucine, based on all proteinaceous matter. In a more preferred embodiment, the composition comprises a protein fraction comprising 43-47 weight % pea protein, 41-45 weight % soy protein and 11 - 13 weight % free leucine, based on all proteinaceous matter. In a preferred embodiment, the sum of said proteins, free leucine and optionally free branched amino acids other than leucine equals 100 weight % of the protein fraction. In a more preferred embodiment, the protein fraction essentially consists of pea protein, soy protein and free leucine. In a preferred embodiment, the composition has no free amino acids, peptides or hydrolysates in the composition, except for free leucine. According to a preferred embodiment, the proteins including soy protein and pea protein are substantially in intact form or non-hydrolysed. In the context of the invention, a "non-hydrolysed" protein is equivalent to an "intact" protein, meaning that the protein has not been subjected to an hydrolysis process. However, minor amounts of hydrolysed proteins may be present in the source of non-hydrolysed proteins, or may be added to the formulation, such as additional amino acids. In a preferred embodiment, the only free amino acids added are branched amino acids leucine, isoleucine and valine. In one embodiment of the present invention, the composition may, in addition to the free leucine, comprise another free amino acid, or a mixture of free amino acids other than leucine, up to about 1 g / 100 ml, preferably up to about 0.5 gram / 100 ml. The protein fraction preferably essentially consists of plant-based proteinaceous matter, in particular proteins, and free leucine. Small amounts of free amino acids other than free leu may be accounted for, preferably less than about 5 %, more preferably less than about 2 %, most preferably less than about 1 % of the protein fraction. In a particularly preferred embodiment, there are no free amino acids (other than leucine), peptides or hydrolysates in the composition.In the context of the invention, the term "about" should preferably be interpreted as a deviation of plus or minus 10 % of the given value.In a preferred embodiment, the composition comprises at least 10 g protein per 100 g dry weight, preferably at least 15 g per 100 g dry weight.In an embodiment, the nutritional composition according to the invention is an oral nutritional supplement. The oral nutritional supplement according to the invention may take any suitable form, such as a pill, drink or powder. In case the composition is in powder form, it is preferably reconstitutable with an aqueous liquid, such as milk or water. The skilled person is able to determine the most suitable form of the composition according to the invention, for example depending on the target group and the further ingredients that may be present.Preferably, the nutritional composition is packaged, stored and provided in a tube feeding bag. Tube feeding is given to provide nutrition to patients which cannot obtain nutrition by swallowing, using a device such as a nasogastric feeding tube or a naso jejunal feeding tube, or by using a percutaneous endoscopic gastrostomy (PEG) or PEG-jejuno-feeding system. The nutritional composition in tube feeding format according to the invention preferably has the form of a complete food, i.e. it can meet all nutritional needs of the user. As such, the nutritional composition according to the invention preferably contains 1000 to 2500 kcal per daily dosage. Depending on the condition of the patient, a daily dose is about 25 to 35 kcal / kg bodyweight / day. Therefore, a typical daily dose for a 70 kg person contains about 2000 kcal. In the context of this application, the state of being fed by a feeding tube is called enteral feeding, comprising all of the abovementioned tube feeding systems, and the nutrition used in the feeding a feeding tube is called enteral nutrition. The (tube feed) composition preferably provides 1.0 -1.8 g protein / kg body weight per day, more preferably 1.0 - 1.8 g protein / kg body weight per day. In an embodiment, the (tube feed) composition comprises, per 100 kcal, 5 - 300 mg taurine, 5 - 300 mg carnitine and 0.5 - 30 mg forthe mixture of low-molecular-weight polyphenols (oligonol), more preferably 10- 300 mg taurine, 10 - 300 mg carnitine and 1 - 30 mg for the mixture of low-molecular-weight polyphenols (oligonol). The tube feeding composition preferably provides 100 - 200 kcal per 100 ml.According to a further embodiment, the composition according to the invention comprises a coenzyme Q10. Coenzyme Q10 (CoQ10) also known as ubiquinone, is a naturally occurring biochemical cofactor (coenzyme) and an antioxidant produced by the human body. It can also be obtained from dietary sources, such as meat, fish, seed oils, vegetable and dietary supplements. CoQ10 is found in may organism, including animals and bacteria. CoQ10 plays a role in mitochondrial oxidative phosphorylation, aiding in the production of adenosine triphosphate (ATP) which is involved in energy transfer within cells. CoQ10 may help to support the skin, brain and lungs as well as protect against chronic diseases like cancer or diabetes. It is known that CoQ10 production decreases with ageing.In a preferred embodiment, the nutritional composition according to the invention comprises 0.4-13 mg / 100 kcal coenzyme Q10.As will be appreciated by the skilled person, the composition may contain further ingredients that are beneficial for the subject. According to a further embodiment, the composition according to the invention optionally comprises one or more of a fat fraction, a carbohydrate fraction, a dietary fibre fraction and micronutrients. Most preferably, the liquid nutritional composition is sterilized or pasteurized. In oneembodiment, the nutritional composition is provided in liquid format, preferably in a ready-to-use liquid form and does not require reconstitution or mixing prior to use. Tube feeding is preferred.ApplicationAccording to the present invention, the combination of taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves is effective in increasing mitochondrial function and supports muscle function. The nutritional composition according to the invention can advantageously be used for prevention and / or treatment of muscle decline of muscle wasting, particularly for improving muscle protein synthesis (MPS). Muscle decline or wasting involves insufficient MPS. In the targeted population where muscle mass, strength and function is compromised, promoting MPS is a therapeutic use. Thus, in one embodiment, the nutritional composition is for use in the treatment and / or prevention of muscle wasting in a human subject.The invention may also be worded in terms of the use of a nutritional composition comprising taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves in the manufacture of a product for prevention and / or treatment of muscle wasting in a human subject. Worded differently, the invention also relates to a method of prevention and / or treatment of muscle wasting in a human subject wherein the subject is administered with a nutritional composition comprising therapeutically effective amounts of taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves. The prophylaxis is particularly directed to a subject at increased risk of muscle wasting, such as described here above.In a preferred embodiment, the invention relates to muscle wasting caused by inflammatory cytokines which in the context of the invention is defined as cytokine-induced muscle wasting.Exercise programIn a preferred embodiment, the human subject receiving the composition of the invention participates in a physical exercise program. The physical exercise regimen or program involves any physical exercise or activity other than or in addition to daily living activities that contributes to a negative energy balance or an activity that costs calories during that activity. Physical activity examples can be, but not limited to, resistance exercise, aerobic exercise or flexibility training or combinations thereof. Endurance training is not recommended, at least not as the sole source of physical exercise, for preserving muscle mass and ameliorating the progression of sarcopenia. Therefore, in one embodiment, the physical exercise regimen does not involve endurance training. It is preferred that the physical exercise regimen involves at least resistance exercise, attributed to its effects on muscle mass maintenance and stimulation of muscle mass increase. Activity patterns could vary from engaging in the above one or more physical activities for a minimum of 1 time per week but preferably 2 times per week and more preferably 3 or more times per week, including at least some form of resistance exercise. There are ACSM / AHA Guidelines for flexibility, endurance and resistance exercise which can be of help to the skilled person to determine a suitable physical exercise regimen, and these are considered incorporatedby reference here. The physical exercise regimen involves daily physical exercise activities as described above, preferably comprising resistance exercise. The physical exercise regimen is preferably in accordance with ACSM / AHA guidelines, their contents herein incorporated by reference.The invention will now be further elucidated by the examples here below, without being limited therebyExample 1C2C12 mouse myoblasts which are the most commonly used cellular model to mimic skeletal muscle in vitro were cultured in culture medium. Culture medium is Dulbecco's Modified Eagle Medium (DMEM) with high glucose (Sigma) and is supplemented with 2.4 g / L NaHCCh, 10 % Foetal Bovine Serum and 1 % Penicillin-Streptomycin. Cells were cultured twice a week until maximal confluency of about 70%. For differentiation into myotubes, cells were plated in Seahorse XFe 96-well plates at 0.5e5cell / mL (80 pL per well). The next day, culture medium was replaced by differentiation medium which is DMEM-high glucose, supplemented with 2.4 g / L NaHCCh, 2 % Horse Serum (heat inactivated) and 1 % Penicillin-Streptomycin. Within 7 days, myoblasts were differentiated into myotubes and exposed to cytokine mixture TNFalpha (10 ng / ml, Fisher 10620924) and Actinomycin D (50 ng / ml, Sigma Aldrich A1410-2MG) for 48 h, and simultaneously incubated with the individual or the combination of the nutritional ingredients in the following final concentrations: 14 pg / ml taurine (111 pM; MW 125 g / mol), 645 pg / ml carnitine (4 mM; MW 161.2 g / mol) and oligonol 1 pg / mL. The reason to treat cells with a trigger (cytokine mixture) is to induce inflammation driven muscle wasting and mimic the clinical situation.Mitochondrial stress test assayThe Agilent Seahorse Cell Mito Stress Test kit is a widely recognized, well accepted standard assay for assessing mitochondrial function. Reference is made to references (1) - (7) using the same or similar. Multiple parameters are obtained in this one assay including, basal respiration, ATP-linked respiration, maximal and reserve capacities, and non-mitochondrial respiration. The assay provides insights into the mechanism of mitochondrial dysfunction and allows users to investigate functional differences among cell types, drug candidates, and genetic or biochemical interventions. The Cell Mito Stress Test Kit contains the compounds, oligomycin, carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP) and rotenone / antimycin A (Rot / A). The Agilent Seahorse Cell Mito Stress Test kit is for use with the Agilent Seahorse XFe96 Analyzer. Using the Agilent Seahorse XFe96 Analyzer, the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of live cells can be measured. This measurement allows for comparing energy engines of mitochondria and glycolysis.The hydrated cartridge was placed in a non-C02 incubator for calibration and after 1 hour, the injection ports were loaded with the compounds, oligomycin, FCCP and Rot / A. Oligomycin (1.5 pM ) inhibits ATP synthase and decreases electron flow through the electron transport chain (ETC) which results in reduced mitochondrial respiration / OCR (decreased cellular ATP production). Subsequent FCCP (1.0pM ) injection disrupts the proton gradient and mitochondrial membrane potential and allows uninhibited electron flow across the ETC, and maximum mitochondrial respiration (increase in OCR). Finally, the combined Rot / A solution (0.5 pM each) was added for effectively shutting down mitochondrial respiration (decrease in OCR). This test allows for the calculation of non-mitochondrial respiration. After compounds were added and mixed, the measurements were done automatically by the Agilent Seahorse XFe96 analyzer in real-time. Output was calculated as spare respiratory capacity which is particularly useful for the intended applications.Statistical analysesAll data are expressed as means±SEM. Statistical analyses were performed using Graphpad Prism, version 10.0.3. Two-way ANOVA followed by Bonferroni post hoc analysis which was used to compare vehicle and trigger. The result of this analysis should be positive, otherwise the experiment was discarded. Least Significant Difference (LSD) post hoc testing was used for the trigger compared to the different nutrient-conditions. Statistical significance is defined as P < 0.05.Results and discussionThe results are shown in terms of spare respiratory capacity which is defined in how much reserve metabolic health the mitochondria have to maintain functioning. As demonstrated in Figure 1 , the individual nutrients only show slight increase in mitochondrial function. However, the combination of taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves showed synergistic effect and spare respiratory capacity was increased to almost 60 % of the vehicle / control / healthy cells -a 1.7 fold increase compared to the muscle wasting condition and almost back to the healthy reference.From these data, it was concluded that the combination of taurine, carnitine and a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves can be used in a nutritional composition to support energy production or cellular nutrition status and prevent / treat muscle wasting, preferably cytokine-induced muscle wasting by stimulating the mitochondrial function.List of references(1) Jiroutkova et al. “Mitochondrial function in skeletal muscle of patients with protracted critical illness and ICU-acquired weakness" Critical Care (2015) 19:448;(2) Tyrrell et al. “Respirometric Profiling of Muscle Mitochondria and Blood Cells Are Associated With Differences in Gait Speed Among Community-Dwelling Older Adults" J. Gerontology series A 70(11) (20150 1394-1399;(3) Li et al. “Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles” J. Physiol. 2016; 594(24) 7197-7213; (4) Park et al “Oxygen Consumption Rates Between Skeletal Muscle Cells Derived from Young and Old Human Donors Elucidate Mitochondrial Dysfunction” Berkeley Scientific Journal fall 2022(5) Fovez et al. “Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients” Cancers 2021 , 13, 6363;(6) Simon et al. “An aerobic exercise intervention to improve metabolic health among people living with HIV with at-risk alcohol use: the ALIVE-Ex research study protocol" AIDS Research and Therapy (2023) 20:35;(7) Bharadwaj et al. “Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy” DOI: 10.3791 / 52350-v.Example 2 - Composition according to the inventionA liquid nutritional product for improving muscle mass, strength and function in an elderly person, the product comprising, per serving of 200 ml, about 21 g proteinaceous matter, with 45 weight % pea protein, 43 weight % soy protein and 12 weight % free leucine, calculated on the weight of the protein fraction. The total amount of leucine is about 19 weight % of all protein. The product has a caloric content of 1.5 kcal / ml, and comprises about 15 weight % carbohydrates, 5.2 weight % fat and 1.4 weight % fibre (31 en% fat, 39 en% carbohydrates, 28 en% protein and 2 en% fibre), minerals and vitamins. The composition comprises 167 mg / 100mL taurine, 167 mg / 100mL carnitine and 16.7 mg / 100mL of Oligonol, i.e. a mixture of low-molecular-weight polyphenols derived from lychee fruit and green tea leaves. The composition is intended to provide 200 mg oligonol per day.

Claims

Claims1. A nutritional composition comprising taurine, carnitine and a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer, wherein the composition comprises, per 100 ml or per 150 kcal, 50 to 300 mg taurine, 50 to 600 mg carnitine and 5 to 30 mg of a mixture of low-molecular-weight polyphenols.

2. A nutritional composition according to claim 1 , wherein the mixture of low-molecular-weight polyphenols comprises 13-18 % flavanol monomers, 14-18 % dimers and 2-6 % trimers.

3. A nutritional composition according to claim 1 or 2, wherein the mixture of low-molecular-weight polyphenols is obtainable by heating lychee fruit with green tea leaves in an acidic aqueous solution.

4. The nutritional composition according to any one of the preceding claims wherein the composition comprises a protein and preferably the protein is selected from whey protein, soy protein, pea protein, canola protein or mixtures thereof.

5. The nutritional composition according to claim 4 wherein the composition comprises at least 10 g protein per 100 g dry weight, preferably at least 15 g per 100 g dry weight.

6. The nutritional composition according to any one of the preceding claims wherein the composition further comprises a coenzyme Q10.

7. The nutritional composition according to any one of the preceding claims wherein the composition comprises, per 100 ml or per 150 kcal, 50 to 300 mg taurine, 100 to 600 mg carnitine and 5 to 30 mg of a mixture of low-molecular-weight polyphenols.

8. The nutritional composition according to any one of the preceding claims, wherein the composition is an oral nutritional supplement or a tube feeding.

9. The nutritional composition according to any one of the preceding claims, for use as a medicament.

10. The nutritional composition according to claims 1 to 9, for use in the treatment and / or prevention of muscle wasting in a human subject.

11. The nutritional composition for use according to claim 10, wherein the human subject is an elderly subject of at least 50 years of age, preferably at least 55 years of age; and / or wherein the human subject is suffering from muscle decline; and / or wherein the human subject is suffering from cancer or sarcopenia or Intensive Care Unit (ICU) acquired weakness.

12. Use of a nutritional composition comprising therapeutically effective amounts of taurine, carnitine and a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer, preferably derived from lychee fruit and green tea leaves, in the manufacture of a product for prevention and / or treatment of muscle wasting in a human subject.

13. A method of prevention and / or treatment of muscle wasting in a human subject comprises administering a nutritional composition comprising therapeutically effective amounts of taurine, carnitine and a mixture of low-molecular-weight polyphenols comprising proanthocyanidin oligomer, preferably derived from lychee fruit and green tea leaves, to the human subject.

14. The nutritional composition according to any one of claims 1 -8, wherein the composition comprises a protein fraction comprising 32-58 weight % pea protein, 30-56 weight % soy protein and 8-16 weight % free leucine, preferably a protein fraction comprising 36-54 weight % pea protein, 34-52 weight % soy protein and 10 - 14 weight % free leucine, and optionally up to 5 weight % of free branched chain amino acids other than leucine, based on all proteinaceous matter, wherein the composition is essentially free from animal protein; wherein the composition preferably comprises a protein fraction comprising 41-50 weight % pea protein, 39-47 weight % soy protein and 10.5 - 13.5 weight % free leucine, based on all proteinaceous matter; wherein the composition preferably comprises a protein fraction comprising 43-47 weight % pea protein, 41-45 weight % soy protein and 11 - 13 weight % free leucine, based on all proteinaceous matter.

15. The nutritional composition according to any one of claims 1 - 8, wherein the composition comprises a protein fraction comprising (a) 15 to 35 weight % of casein ; (b) 25 to 50 weight % of whey protein ; (c) 10 to 30 weight % of soy protein ; and (d) 10 to 30 weight % of pea protein, relative to the total protein in the protein fraction, wherein the sum of said proteins preferably equals 100 weight % ofthe protein fraction; wherein the composition preferably comprises a protein fraction comprising (a) 20 to 30 weight % of casein ; (b) 30 to 40 weight % of whey protein ; (c) 15 to 25 weight % of soy protein ; and (d) 15 to 25 weight % of pea protein, relative to the total protein in the protein fraction, wherein the sum of said proteins preferably equals 100 weight % ofthe protein fraction.

Images