Composition for use in increasing spermidine production by microbiota

Abstract

The present invention relates to compositions comprising at least one source of protein and at least one source of prebiotic for use in increasing spermidine production by the gut microbiota. The composition can be used in preventing and / or treating age-related disorders.

Description

[0001] COMPOSITION FOR USE IN INCREASING SPERMIDINE PRODUCTION BY MICROBIOTA

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to compositions comprising at least one source of protein and at least one source of prebiotic for use in increasing spermidine production by the gut microbiota. The composition can be used in preventing and / or treating age-related disorders.

[0004] BACKGROUND OF THE INVENTION

[0005] Polyamines are small polycationic molecules with a wide array of biological functions including gene regulation, stress resistance, cell proliferation and differentiation, and are associated to both eukaryotic and prokaryotic cells.

[0006] Spermidine, putrescine and spermine are the dominate polyamines in eukaryotes. These polyamines regulate important cellular functions, including growth and proliferation, RNA and DNA stability, RNA-to-protein translation, autophagy and immune responses.

[0007] Among them, spermidine has emerged as an anti-ageing metabolite that is vital for cellular and tissue function in different organisms including humans. Several lines of evidence support its anti-ageing properties: spermidine levels decrease in most investigated tissues and species during aging1; spermidine supplementation elongates lifespan and body functions of multiple species1; epidemiological data support the notion that elevated dietary intake alleviates, delays or halts age-associated deteriorations, including cancer, cardiovascular disease, cognitive dysfunction2-4, compromised intestine barrier integrity5-6or leaky gut, compromised gut microbiome, which are contributing factors to age-associated inflammation7.

[0008] The ingested food, such as soybeans, mushrooms, wheat germ, beef, pork, chicken livers, oranges, turban shell viscera, and green tea leaves, is the major direct source of spermidine in the lumen.

[0009] However, the upper parts of intestine adsorb most of these compounds. Thus, direct administration of spermidine sources has limited effects on the level of spermidine in circulation8.

[0010] The gut microbiota is considered the main responsible for spermidine levels in the lower part of intestine. Gut bacteria synthesize polyamines by using arginine, ornithine or lysine as a substrate. Polyamines, including spermidine, produced in the colonic lumen are transferred into the bloodstream via the colonic mucosa, which may influence certain cellular function in tissue or cell specific manner that leads to anti-ageing effects.

[0011] On average, 3 to 18 g of dietary escape from digestion and absorption in the small intestine and enter the colon. Bacteria in the colon can further break them down to small peptides and free amino acids which can be further metabolized to compounds such as polyamines, including spermidine.

[0012] However, there is no study comparing the efficacy of different dietary nutrients on microbial production of spermidine.

[0013] Thus, there is a clear unmet need for new compositions that increase spermidine production by the gut microbiota and, as result, increase resistance to age-related pathologies, improve a physiological state or disorder related to cell ageing, improve a physiological state linked to metabolic fatigue in one or more cells, increase mitochondrial energy in one or more cells, increase antioxidant capacity, reduce oxidative stress and / or enhance mitochondrial function, improve mobility and / or improve healthspan and / or lifespan in a subject.

[0014] References

[0015] 1. Hofer, S. J.; Simon, A. K.; Bergmann, M.; Eisenberg, T.; Kroemer, G.; Madeo, F., Mechanisms of spermidine-induced autophagy and geroprotection. Nature Aging 2022, 2 (12), 1112-1129.

[0016] 2. Kiechl, S.; Pechlaner, R.; Willeit, P.; Notdurfter, M.; Paulweber, B.; Willeit, K.; Werner, P.; Ruckenstuhl, C.; Iglseder, B.; Weger, S.; Mairhofer, B.; Gartner, M.; Kedenko, L.; Chmelikova, M.; Stekovic, S.; Stuppner, H.; Oberhollenzer, F.; Kroemer, G.; Mayr, M.; Eisenberg, T.; Tilg, H.; Madeo, F.; Willeit, J., Higher spermidine intake is linked to lower mortality: a prospective population-based study. The American Journal of Clinical Nutrition 2018, 108 (2), 371-380.

[0017] 3. Schroeder, S.; Hofer, S. J.; Zimmermann, A.; Pechlaner, R.; Dammbrueck, C.; Pendl, T.; Marcello, G. M.; Pogatschnigg, V.; Bergmann, M.; Muller, M.; Gschiel. V.; Ristic, S.; Tadic, J.; Iwata, K.; Richter, G.; Farzi, A.; Ugal, M.; Schafer, U.; Poglitsch, M.; Royer, P.; Mekis, R.; Agreiter, M.; Tolle, R. C.; Sotonyi, P.; Willeit, J.; Mairhofer, B.; Niederkofler, H.; Pallhuber, I.; Rungger, G.; Tilg, H.; Defrancesco, M.; Marksteiner, J.; Sinner, F.; Magnes, C.; Pieber, T. R.; Holzer, P.; Kroemer, G.; Carmona-Gutierrez, D.; Scorrano, L.; Dengjel, J.; Madl, T.; Sedej, S.; Sigrist, S. J.; Racz, B.; Kiechl, S.; Eisenberg, T.; Madeo, F., Dietary spermidine improves cognitive function. Cell Reports 2021, 35 (2) 108985.

[0018] 4. Wu, H.; Wang, J.; Jiang, H.; Liu, X.; Sun, X.; Chen, Y.; Hu, C.; Wang, Z.; Han, T.; Sun, C.; Wei, W.; Jiang, W., The association of dietary spermidine with all-cause mortality and CVD mortality: The U.S. National Health and Nutrition Examination Survey, 2003 to 2014. Frontiers in Public Health 2022, 10:949170.

[0019] 5. Ma L, Ni Y, Wang Z, Tu W, Ni L, Zhuge F, Zheng A, Hu L, Zhao Y, Zheng L, Fu Z. Spermidine improves gut barrier integrity and gut microbiota function in diet-induced obese mice. Gut Microbes. 2020 Nov 9; 12(1): 1 -19. doi: 10.1080 / 19490976.2020.1832857. PMID: 33151120; PMCID: PMC7668533. 6. Jacopo J.V. Branca, Massimo Gulisano, Claudio Nicoletti, Intestinal epithelial barrier functions in ageing, Ageing Research Reviews, Volume 54, 2019, 100938, ISSN 1568-1637.

[0020] 7. Lan Zhang, Junbin Yan, Chi Zhang, Shuyan Feng, Zheli Zhan, Yang Bao, Shuo Zhang, Guanqun Chao, Improving intestinal inflammaging to delay aging? A new perspective, Mechanisms of Ageing and Development, Volume 214, 2023, 111841 , ISSN 0047-6374.8.

[0021] Senekowitsch, S.; Wietkamp, E.; Grimm, M.; Schmelter, F.; Schick, P.; Kordowski, A.; Sina, C.; Otzen, H.; Weitschies, W.; Smollich, M., High-Dose Spermidine Supplementation Does Not Increase Spermidine Levels in Blood Plasma and Saliva of Healthy Adults: A Randomized Placebo-Controlled Pharmacokinetic and Metabolomic Study. Nutrients 2023, 75 (8), 1852.

[0022] SUMMARY OF THE INVENTION

[0023] The present invention provides composition increasing spermidine production by the gut microbiota.

[0024] In particular, the present invention provides composition comprising at least one source of protein and at least one source of prebiotic for use in increasing spermidine production by the gut microbiota.

[0025] In an embodiment, the composition comprises the source of protein that is at least one selected from the group: pulses, grains, nuts, algae, oil crop and / or its press cakes, animal proteins or combination thereof.

[0026] In an embodiment, pulses are at least one selected from the group comprising faba bean, chick pea, green pea, lentil.

[0027] In an embodiment, grains are at least one selected from the group comprising rice, oat, quinoa, wheat, barley.

[0028] In an embodiment, nuts are at least one selected from walnut, almond, pistachio.

[0029] In an embodiment, algae are at least one selected from seaweed, microalgae.

[0030] In an embodiment, oil crop and / or its press cakes are selected from soy, canola, peanut, bambara.

[0031] In an embodiment, animal proteins are at least one selected from milk, egg, seafood, poultry, red meat, white meat.

[0032] In an embodiment, the composition comprises the source of prebiotic that is at least one oligosaccharide. In an embodiment, the composition comprises the at least one oligosaccharide that is selected from the group: fructo-oligosaccharide, inulin, guar gum, galactooligosaccharide, betaglucan, resistant starch, arabinoxylan, pectin, raffinose, polydextrose, agar, alginate or combination thereof.

[0033] In an embodiment, the composition comprises the at least one source of protein and the at least one source of prebiotic in an effective amount.

[0034] In an embodiment, the composition comprises the source of protein that is administered to the subject in an amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.

[0035] In an embodiment, the composition comprises the source of prebiotic that is administered to the subject in an amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

[0036] In an embodiment, the composition comprises the at least one source of protein that is faba bean and the at least one source of prebiotic that is fructo-oligosaccharide.

[0037] In an embodiment, the composition comprises:

[0038] - faba bean protein in the amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.

[0039] - fructo-oligosaccharide in the amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

[0040] In an embodiment, the composition comprises the at least one source of protein that is pulses protein and the at least one source of prebiotic that is fructo-oligosaccharide.

[0041] In an embodiment, the composition comprises:

[0042] - pulses protein in the amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.

[0043] - fructo-oligosaccharide in the amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

[0044] In an embodiment, the composition is for use in preventing and / or treating age-related disorders. In an embodiment, the age-related disorder is at least one selected from the group: a physiological state or disorder related to cell ageing, a physiological state linked to metabolic fatigue in one or more cells, excessive catabolism, oxidative stress, reduced antioxidant capacity and / or mitochondrial function, impaired mobility, healthspan and / or lifespan in a subject, muscle dysfunction, including sarcopenia, frailty, inclusion body myositis, myopathy / myolysis induced by drugs such as corticosteroids or statins, muscle wasting induced by immobilization or hospitalization, muscle weakness, immune function disorders, cognitive function disorders, including delirium, dementia, learning disorder, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD), cardiovascular disease, compromised intestine barrier integrity or leaky gut, compromised gut microbiome, age-associated inflammation.

[0045] In an embodiment, the composition is for use in: increasing (i) resistance to age-related pathologies, (ii) improving a physiological state or disorder related to cell ageing, (iii) improving a physiological state linked to metabolic fatigue in one or more cells, (iv) increasing mitochondrial energy in one or more cells, (v) increasing antioxidant capacity, reducing oxidative stress and / or enhancing mitochondrial function, (vi) improving mobility and / or (vii) improving healthspan and / or lifespan in a subject.

[0046] In an embodiment, the composition is in the form of food composition, dietary supplement, nutritional composition, nutraceutical, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, medicament, beverage and drink, including coffee-based product, oral nutritional supplement (ONS), a complete nutritional formula, a pharmaceutical, a medical or a food product.

[0047] In an embodiment, the subject is a human, a farm animal or a pet.

[0048] Another aspect of the present disclosure is use of the composition in the subject in need of at least one increasing (i) resistance to age-related pathologies, (ii) improving a physiological state or disorder related to cell ageing, (iii) improving a physiological state linked to metabolic fatigue in one or more cells, (iv) increasing mitochondrial energy in one or more cells, (v) increasing antioxidant capacity, reducing oxidative stress and / or enhancing mitochondrial function, (vi) improving mobility and / or (vii) improving healthspan and / or lifespan in an individual.

[0049] Benefits from this improvement are in preventing and / or treating age-related disorders.

[0050] Benefits from this improvement also include (i) increasing resistance to age-related pathologies, (ii) improving a physiological state or disorder related to cell ageing, (iii) improving a physiological state linked to metabolic fatigue in one or more cells, (iv) increasing mitochondrial energy in one or more cells, (v) increasing antioxidant capacity, reducing oxidative stress and / or enhancing mitochondrial function, (vi) improving mobility and / or (vii) improving healthspan and / or lifespan in a subject.

[0051] Additional features and advantages are described herein and will be apparent from the following Figures and Detailed Description.

[0052] DESCRIPTION OF FIGURES

[0053] Figure 1. Effects of protein sources and prebiotic FOS on microbial production of spermidine using an in vitro fermentation system from Example 1.

[0054] Proteins were digested following INFOGEST protocol, and small peptides and free amino acids were removed with dialysis process using a 14 kD molecular weight cut off. In vitro fermentation was performed with three fecal samples and spermidine concentrations at 24 hours are shown in (A). FOS was added to the fermentation media and the results are shown in (B). Data are mean + / - sem. Statistical analysis was performed with one way ANOVA followed by Tukey post hoc analysis. No shared letter between groups indicated statistically significant difference

[0055] Figure 2. The effect of FOS on the production of spermidine (A) and putrescine (B) by the gut microbiota from Example 1.

[0056] Procedures of the experiments followed the description in the Methods and Materials for Example 1 in Examples. In an upper and lower gut combine in vitro system. Data are mean + / - sem, n=3. Effect of FOS for each protein source was tested by Student t-test, ** p<0.01 , * p<0.05, § p< 0.1 . (p<0.05 in post hoc analysis).

[0057] Figure 3. The relationship between arginine in proteins and the production of spermidine in a time course study using in vitro fermentation experiment from Example 1.

[0058] Experimental conditions are shown in the Methods and Materials for Example 1 in Examples. The amount of arginine was obtained from the analytical report or production description from the manufacturers. Fermentation media were collected at 6, 24 and 48h for polyamine analysis, for Linear regression analysis was performed for 6h (A), 24h (B) and 48h (C) separately, and p-values and R2 are shown below each graph. Figure 4. Microbial production of spermidine (A) and putrescine (B) in an in vitro fermentation experiment from Example 2.

[0059] Data are mean + / - sem from an experiment with three different microbiota. Data were normalized to the amount of enzyme only and were expressed as a ratio to enzyme only. Results were analyzed with one way ANOVA test followed by Tukey posthoc analysis. Letters not shared between groups indicate statistical significance (p<0.05).

[0060] DATAILED DESCRIPTION

[0061] Definitions

[0062] Some definitions are provided hereafter. Nevertheless, definitions may be located in the “Embodiments” section below, and the above header “Definitions” does not mean that such disclosures in the “Embodiments” section are not definitions.

[0063] As used in this disclosure and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a vitamin” or “the vitamin” encompass both an embodiment having a single vitamin and an embodiment having two or more vitamins.

[0064] The words “comprise,” “comprises” and “comprising” are to be interpreted inclusively rather than exclusively. Likewise, the terms “include,” “including” and “or” should all be construed to be inclusive, unless such a construction is clearly prohibited from the context. Nevertheless, the compositions disclosed herein may lack any element that is not specifically disclosed herein. Thus, a disclosure of an embodiment using the term “comprising” includes a disclosure of embodiments “consisting essentially of” and “consisting of’ the components identified.

[0065] The terms “at least one of” and “and / or” used in the respective context of “at least one of X or Y” and “X and / or Y” should be interpreted as “X,” or “Y,” or “X and Y.” For example, “at least one of resistance or recovery” and “resistance and / or recovery” should be interpreted as “resistance,” or “recovery,” or “both resistance and recovery.”

[0066] Where used herein, the terms “example” and “such as,” particularly when followed by a listing of terms, are merely exemplary and illustrative and should not be deemed to be exclusive or comprehensive. As used herein, a condition “associated with” or “linked with” another condition means the conditions occur concurrently, preferably means that the conditions are caused by the same underlying condition, and most preferably means that one of the identified conditions is caused by the other identified condition.

[0067] A “subject” is a mammal, preferably a human, a farm animal, a pet. In particular the term “farm animal” may include but is not limited to a horse (e.g., a pet or horse undergoing medical treatment), or cattle or poultry (e.g., cattle or poultry being used in agriculture). In particular the term “pet” means any animal which could benefit from or enjoy the compositions provided by the present disclosure. For example, the pet can be an avian, bovine, canine, equine, feline, hircine, lupine, murine, ovine, or porcine animal, but the pet can be any suitable animal.

[0068] “Prevention” includes reduction of risk, incidence and / or severity of a condition or disorder. The terms “treatment” and “treat” include both prophylactic or preventive treatment (that prevent and / or slow the development of a targeted pathologic condition or disorder) and curative, therapeutic or disease-modifying treatment, including therapeutic measures that cure, slow down, lessen symptoms of, and / or halt progression of a diagnosed pathologic condition or disorder; and treatment of patients at risk of contracting a disease or suspected to have contracted a disease, as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition.

[0069] The terms “treatment” and “treat” do not necessarily imply that a subject is treated until total recovery. The terms “treatment” and “treat” also refer to the maintenance and / or promotion of health in an individual not suffering from a disease but who may be susceptible to the development of an unhealthy condition. The terms “treatment” and “treat” are also intended to include the potentiation or otherwise enhancement of one or more primary prophylactic or therapeutic measures. As non-limiting examples, a treatment can be performed by a patient, a caregiver, a doctor, a nurse, or another healthcare professional.

[0070] As used in this disclosure and the appended claims, the term “upper gastrointestinal tract” refers to the gastrointestinal tract comprising the mouth, pharynx, esophagus, stomach, and duodenum of the small intestine.

[0071] As used in this disclosure and the appended claims, the term “lower gastrointestinal tract” refers the gastrointestinal tract comprising the remainder of the small intestine, i.e., the jejunum and ileum, and all of the large intestine, i.e., the cecum, colon, rectum, and anal canal. Bacteria can be found in the gastrointestinal tract, and particularly in the intestines. As used herein, a prophylactically or therapeutically “effective amount” is an amount that prevents a deficiency, treats a disease or medical condition in an individual, or, more generally, reduces symptoms, manages progression of the disease, or provides a nutritional, physiological, or medical benefit to the individual.

[0072] As used herein, the terms “food,” “food product” and “food composition” mean a product or composition that is intended for oral ingestion by a human or other mammal and comprises at least one nutrient for the human or other mammal.

[0073] “Nutritional compositions” and “nutritional products,” as used herein, include any number of food ingredients and possibly optional additional ingredients based on a functional need in the product and in full compliance with all applicable regulations. It may refer to liquids, powders, gels, pastes, solids, concentrates, suspensions, or ready-to-use forms of enteral formulas, oral formulas, formulas for children, formulas for adults, porridges and / or cereals, food products, food compositions, baby food, pet food. The optional ingredients may include, but are not limited to, conventional food additives, for example one or more, acidulants, additional thickeners, buffers or agents for pH adjustment, chelating agents, colorants, emulsifies, excipient, flavor agent, mineral, osmotic agents, a pharmaceutically acceptable carrier, preservatives, stabilizers, sugar, sweeteners, texturizers, and / or vitamins. The optional ingredients can be added in any suitable amount.

[0074] “Prebiotic” generally means a non-digestible food ingredient that beneficially affects the host by selectively stimulating the growth and / or activity of micro-organisms present in the gut of the host, and thus attempts to improve host health.

[0075] The term “oligosaccharide” means a carbohydrate having a degree of polymerization (DP) ranging from 2 to 20 inclusive but not including lactose. In some embodiments of the invention, carbohydrate has DP ranging from 3 to 20.

[0076] The expressions “galacto-oligosaccharide” and “GOS” can be used interchangeably. They refer to an oligosaccharide comprising two or more galactose molecules which has no charge and no N-acetyl residue (i.e. they are neutral oligosaccharide).

[0077] The expressions “fructo-oligosaccharides” and “FOS” can be used interchangeably. They refer to an oligosaccharide comprising short fructose chains.

[0078] The expression “guar gum” refers to a galactomannan polysaccharide comprised of galactose and mannose. The expression “beta-glucan” refers to a polysaccharide comprised of beta-D-glucose.

[0079] The expression “resistant starch” refers to a polysaccharide comprised of amylose and amylopectin.

[0080] The expression “arabinoxylan” refers to a hemicellulose comprised of arabinose and xylose.

[0081] The expression “pectin” refers to a polysaccharide comprised of mainly galacturonic acid and other less abundant sugar residues xylose, apiose, rhamnose, galactose, arabinose.

[0082] The expression “raffinose” refers to a trisaccharide comprised of galactose, glucose and fructose.

[0083] The expression “polydextrose” refers to a synthetic polysaccharide comprised of glucose.

[0084] The expression “agar” refers to a polysaccharide comprised of agarose and agaropectin.

[0085] The expression “alginate” refers to polysaccharide comprised of beta-D-mannuronate and alpha-L-guluronate.

[0086] The expressions “oligosaccharide”, “oligosaccharides”, “oligosaccharide mixture” or “mixture of oligosaccharide(s)” can be used interchangeably. In some advantageous embodiments the oligosaccharides of the oligosaccharide mixture are bovine milk oligosaccharides, bovine milk-derived oligosaccharides, or oligosaccharides derived from bovine milk (all also referred to as “BMOs”).

[0087] As used herein, "cognitive function" refers to any mental process that involves symbolic operations, e.g., perception, memory, attention, speech comprehension, speech generation, reading comprehension, creation of imagery, learning, and reasoning, preferably at least memory. Methods for measuring cognitive function are well-known and can include, for example, individual or battery tests for any aspect of cognitive function. One such test is the Prudhoe Cognitive Function Test by Margallo-Lana et al. (2003) J. Intellect. Disability Res. 47:488-492. Another such test is the Mini Mental State Exam (MMSE), which is designed to assess orientation to time and place, registration, attention and calculation, recall, language use and comprehension, repetition, and complex commands. As used herein, a “cognitive disorder” refers to any condition that impairs cognitive function. Non-limiting examples of a cognitive disorder include delirium, dementia, learning disorder, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD). A "stress-induced or stress-related cognitive dysfunction" refers to a disturbance in cognitive function that is induced or related to stress.

[0088] The term "unit dosage form," as used herein, refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of the composition disclosed herein in an amount sufficient to produce the desired effect, in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for the unit dosage form depend on the particular compounds employed, the effect to be achieved, and the pharmacodynamics associated with each compound in the host.

[0089] The “subject” or “individual” of the present invention is a human adult subject, preferably a healthy adult, an animal, a pet with the need to improve motivational and / or mental performance through modulating glutathione levels in the brain. The compositions of the invention may be beneficially used for increasing glutathione level in the brain, for preventing or treating conditions or diseases which are characterized by low glutathione levels in the brain, whether transient or chronic.

[0090] EMBODIMENTS

[0091] The inventors surprisingly found that a combination of specific sources of proteins and prebiotics promote spermidine production by the gut microbiota.

[0092] The inventors tested the concept of dietary proteins as a source for microbial polyamine production, in particular spermidine using an in vitro digestion and fermentation platform. Example 1 was performed by ProDigest showed that protein alone is not enough to stimulate gut microbiota to produce spermidine. Microbiota requires carbon sources such FOS to produce spermidine. More importantly, the effect of FOS was observed with spermidine but not with putrescine, suggesting that some but not entire microbial polyamine synthesis pathway agmatine is sensitive to the prebiotic fermentation. Moreover, addition of carbon source FOS extended the microbial production of spermidine beyond 6 hours. This further highlight the cross talk between polyamine synthesis and fiber fermentation in the gut ecosystem.

[0093] In the Example 1 , the inventors found that faba bean protein was very efficient in increasing spermidine over the control group, where the concentrations of putrescine were similar in all groups. The faba bean protein concentrate tested in the current Example 1 contains 40% (w / w) of non-protein materials. Thus, an aspect of the present disclosure is a composition comprising at least one source of protein and at least one source of prebiotic for use in increasing spermidine production by the gut microbiota of a subject.

[0094] In an embodiment, the composition comprises the source of protein that is at least one selected from the group: pulses, grains, nuts, algae, oil crop and / or its press cakes, animal proteins or combination thereof.

[0095] In an embodiment, pulses are at least one selected from the group comprising faba bean, chick pea, green pea, lentil.

[0096] In an embodiment, grains are at least one selected from the group comprising rice, oat, quinoa, wheat, barley.

[0097] In an embodiment, nuts are at least one selected from walnut, almond, pistachio.

[0098] In an embodiment, algae are at least one selected from seaweed, microalgae.

[0099] In an embodiment, oil crop and / or its press cakes are selected from soy, canola, peanut, bambara.

[0100] In an embodiment, animal proteins are at least one selected from milk, egg, seafood, poultry, red meat, white meat.

[0101] Suitable protein sources may be selected from any suitable animal or plant protein source.

[0102] In an embodiment, the composition comprises the source of prebiotic that is at least one oligosaccharide.

[0103] The oligosaccharides can be isolated from any source. Preferably the oligosaccharides are isolated, purified or concentrated from the protein sources. Alternatively, all or some of the oligosaccharides are produced in totality or in part by bioengineering.

[0104] In an embodiment, the composition comprises the at least one oligosaccharide that is selected from the group: fructo-oligosaccharide, inulin, guar gum, galactooligosaccharide, betaglucan, resistant starch, arabinoxylan, pectin, raffinose, polydextrose, agar or combination thereof.

[0105] In an embodiment, the composition comprises the at least one source of protein and the at least one source of prebiotic in an effective amount. In an embodiment, the composition comprises the source of protein that is administered to the subject in an amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.

[0106] In an embodiment, the composition comprises the source of prebiotic that is administered to the subject in an amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

[0107] In an embodiment, the composition comprises the at least one source of protein that is faba bean and the at least one source of prebiotic that is fructo-oligosaccharide.

[0108] In an embodiment, the composition comprises:

[0109] - faba bean protein in the amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.

[0110] - fructo-oligosaccharide in the amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

[0111] In an embodiment, the composition comprises the at least one source of protein that is pea and the at least one source of prebiotic that is fructo-oligosaccharide.

[0112] In an embodiment, the composition comprises the at least one source of protein that is pulses and the at least one source of prebiotic that is fructo-oligosaccharide.

[0113] In an embodiment, the composition comprises:

[0114] - Pulses protein in the amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.

[0115] - fructo-oligosaccharide in the amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

[0116] In an embodiment, the composition is for use in preventing and / or treating age-related disorders.

[0117] In an embodiment, the age-related disorder is at least one selected from the group: a physiological state or disorder related to cell ageing, a physiological state linked to metabolic fatigue in one or more cells, excessive catabolism, oxidative stress, reduced antioxidant capacity and / or mitochondrial function, impaired mobility, healthspan and / or lifespan in a subject, muscle dysfunction, including sarcopenia, frailty, inclusion body myositis, myopathy / myolysis induced by drugs such as corticosteroids or statins, muscle wasting induced by immobilization or hospitalization, muscle weakness, immune function disorders, cognitive function disorders, including delirium, dementia, learning disorder, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD), cardiovascular disease, compromised intestine barrier integrity or leaky gut, compromised gut microbiome, age-associated inflammation.

[0118] In an embodiment, the composition is for use in: increasing (i) resistance to age-related pathologies, (ii) improving a physiological state or disorder related to cell ageing, (iii) improving a physiological state linked to metabolic fatigue in one or more cells, (iv) increasing mitochondrial energy in one or more cells, (v) increasing antioxidant capacity, reducing oxidative stress and / or enhancing mitochondrial function, (vi) improving mobility and / or (vii) improving healthspan and / or lifespan in a subject.

[0119] In an embodiment, the composition is in the form of food composition, dietary supplement, nutritional composition, nutraceutical, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, medicament, beverage and drink, including coffee-based product, oral nutritional supplement (ONS), a complete nutritional formula, a pharmaceutical, a medical or a food product.

[0120] In an embodiment, the subject is a human, a farm animal or a pet.

[0121] Another aspect of the present disclosure is use of the composition in the subject in need of at least one increasing (i) resistance to age-related pathologies, (ii) improving a physiological state or disorder related to cell ageing, (iii) improving a physiological state linked to metabolic fatigue in one or more cells, (iv) increasing mitochondrial energy in one or more cells, (v) increasing antioxidant capacity, reducing oxidative stress and / or enhancing mitochondrial function, (vi) improving mobility and / or (vii) improving healthspan and / or lifespan in an individual.

[0122] An advantage of one or more embodiments provided by the present disclosure is to improve the condition of subjects, animals, humans, ageing animals, or ageing humans.

[0123] Still another advantage of one or more embodiments provided by the present disclosure is to reduce or prevent the risk of morbidity or mortality due to excessive catabolism.

[0124] An additional advantage of one or more embodiments provided by the present disclosure is to protect an ageing subject from muscle dysfunction, for example sarcopenia, frailty, inclusion body myositis, myopathy / myolysis induced by drugs such as corticosteroids or statins, muscle wasting induced by immobilization or hospitalization. An additional advantage of one or more embodiments provided by the present disclosure is to protect an ageing subject from muscle weakness.

[0125] Another advantage of one or more embodiments provided by the present disclosure is to protect an aging subject from cardiovascular disease or condition.

[0126] Another advantage of one or more embodiments provided by the present disclosure is to protect an aging subject from compromised intestine barrier integrity or leaky gut, compromised gut microbiome, age-associated inflammation.

[0127] Still another advantage of one or more embodiments provided by the present disclosure is to increase the survivability of a critically ill patient or an ageing subject.

[0128] An additional advantage of one or more embodiments provided by the present disclosure is to accelerate the regain of mobility, or shorten the time of immobility, after discharge from the intensive care unit.

[0129] The present combination of ingredient is also effective in improving healthspan. Moreover, increasing mean lifespan is an indication for improved healthspan as shown in Martineau CN, Brown AEX, Laurent P (2020) PLoS Comput Biol 16(7).

[0130] Reference is made hereinafter in detail to specific embodiments of the invention. While the invention will be described in conjunction with these specific embodiments, it will be understood that it is not intended to limit the invention to such specific embodiments. On the contrary, it is intended to cover alternatives, modifications, and equivalents as may be included within the spirit and scope of the invention as defined by the claims. Numerous specific details are set forth in the description in order to provide a thorough understanding of the present invention. The present invention may be practiced without some or all of these specific details. In other instances, well known methods and protocols have not been described in detail, in order not to unnecessarily obscure the present invention.

[0131] Composition formulations

[0132] In one embodiment, the compositions are food compositions, including human and pet food compositions.

[0133] For pet food compositions, they may supply the necessary dietary requirements for an animal, animal treats (e.g., biscuits), or dietary supplements. The compositions may be a dry composition (e.g., kibble), semi-moist composition, wet composition, or any mixture thereof. In another embodiment, the composition is a dietary supplement such as a gravy, drinking water, beverage, yogurt, powder, granule, paste, suspension, chew, morsel, treat, snack, pellet, pill, capsule, tablet, or any other suitable delivery form. The dietary supplement is to be administered in small amounts, or in the alternative, can be diluted before administration. The dietary supplement may require admixing or can be admixed with water or other diluent prior to administration.

[0134] In another embodiment, the composition is in the form of a food supplement or dietary supplement, in particular in the form of a tablet, a powder sachet or a gummy.

[0135] In a preferred embodiment, the composition is in the form of an effervescent tablet.

[0136] The effervescent tablets can be produced and controlled same as conventional tablets. These controls include physicochemical properties such as hardness, weight variation, friability, solution time, pH and content uniformity. The effervescent tablets can be produced by a direct compression method, a fusion method, a wet or dry granulation method, or any other suitable method. Low relative humidity (e.g., maximum of 25% or less) and moderate to cool temperatures (e.g., about 25 °C or 77 °F) in the environment may be essential to prevent sticking granule or tablets to the tablet press machine.

[0137] In the direct compression method, the effervescent tablet can be formed by compressing the ingredients in the form of powders into a dense mass, for example, by a tablet press machine. The powdered ingredients may be first granulized to similar or equal sizes before being made into tablets, so that the mixtures of powder have excellent flowability without particles segregation. Granulating may not be required if the raw materials are selected to achieve a free-flowing, nonsegregating, compressible powder blend. The tablets can then be dried by heat, such as in an oven with air circulation, at a suitable temperature for a suitable time and after cooling can be packed in a suitable package.

[0138] In the fusion method, the ingredients can be mixed in a suitable mixer, such as a blender, for an appropriate time. Then, the obtained mixture can be heated to a suitable temperature. The powder may be mixed regularly until the crystallization water of citric acid is released as binder factor (e.g., approximately 30 minutes) and an appropriate pasty mass is obtained. This wet mass can be passed through a sieve to obtain the desired granules, which can then be dried at a suitable temperature for an appropriate time. After drying, the granules can be passed through the sieve again. Other ingredients can be added to the granule mass and mixed for a suitable time. The granule mixtures then can be compressed into tablets by the tablet press machine. Finally, the tablets can be dried and packed in a suitable package.

[0139] In the wet granulation method, the ingredients may be milled by a miller, either separately or as a mixture with ethanol, ethanol-water mixture, isopropanol, etc., and the obtained powder can be passed through a sieve and then blended. A binder solution can be added to the mixture to form a pasty mass. This pasty mass can then be passed through a sieve to obtain desired granules, which can then dried. The dried mass can be passed through a sieve again, and other ingredients can be added and mixed. The obtained granule mixtures then can be compressed into tablets by the tablet press machine. Finally, the tablets can be dried and packed in a suitable package. Wet granulation can also be performed by carefully adding 0.1 to 1.0 per cent water (weight-to-weight basis) to a blend of raw materials that possess the uniformity, compressibility, and flowability needed to produce good-quality tablets, but which lacks the needed binding properties. The free water which is usually added in the form of a fine spray to selected formulation components while mixing in a suitable blender acts as a binder. The granulation steps must be precisely timed and the ingredients mixed thoroughly to distribute the granulating fluid evenly in the blend. The mix is then quickly discharged to drying ovens. After drying, the granules are sized, and a final mix is performed. The granules are then compressed into tablets using tablet machines.

[0140] The dry granulation method can use special processing equipment known as a “roller compactor” or “chilsonator.” These machines compress premixed powders between two counterrotating rollers under extreme pressure. Depending on the configuration of the roller, the feed material may be compacted into dense ribbon-like materials known as flakes (smooth rolls) or dense briquettes (almond or stick-shaped) if the rollers have grooved or etched surfaces. The compressed material is reduced to the proper size for tablet granulation purposes. Another dry granulation procedure is slugging, in which the powder particles are compressed into large flat tablets or pellets using a tablet press or more usually, a heavy-duty tablet-compacting equipment. The resulting tablet or slug are milled to yield the desired granule characteristics.

[0141] The effervescent tablet can be made in any shape and can have any suitable size. As non-limiting examples, the tablet can be 5 mm to 20 mm long, thick, and / or in diameter if having a round shape. The size of the tablet can be from 5 mm to 10 mm, 5 mm to 15 mm, 10 mm to 15 mm, 10 mm to 20 mm, or 15 mm to 20 mm.

[0142] Y1 The effervescent tablet may comprise a binder, such as polyvinylpyrolidone (PVP) or any other suitable binder. The binder is preferably water-soluble. It can be added as dry powder or in a wet form as an aqueous or hydroalcoholic solution. Mannitol, PEG 6000 and water in small amounts can also be used as a binder. PEG 6000 at 3% use level can be used as a dry binder. The ideal amount of binder is one that makes the tablet hard enough to handle but soft enough to disintegrate and dry enough to be stable. The effervescent tablet may also be formulated without a binder.

[0143] Beverage compositions

[0144] In one embodiment, the compositions are beverage compositions. Such beverage compositions are meant to be consumed by a human or animal. In several embodiments, the beverage is a milk-based beverage; a performance nutrition product, a medical nutrition product; a milk product, e.g. a milk drink, a product with at least one nutrient for improving motivation performance or mental energy.

[0145] Dairy product

[0146] In one embodiment, the composition can be formulated as a “dairy product” together with milk proteins, e.g., milk protein concentrate or milk protein isolate; caseinates or casein, e.g., micellar casein concentrate or micellar casein isolate; or whey protein, e.g., whey protein concentrate or whey protein isolate.

[0147] Nutritional supplement

[0148] In one embodiment, the composition of the invention can be formulated as a “nutritional supplement” together with glutathione enhancing compounds of the invention. The compounds of the invention can be used alone or in combination with appropriate additives to make tablets, gummies, powders, granules or capsules, for example, with conventional additives, such as lactose, mannitol, corn starch or potato starch; with binders, such as crystalline cellulose, cellulose functional derivatives, acacia, corn starch or gelatins; with disintegrators, such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants, such as talc or magnesium stearate; and if desired, with diluents, buffering agents, moistening agents, preservatives and flavoring agents.

[0149] Administration

[0150] The composition of the invention can be administered at least one day per week, preferably at least two days per week, more preferably at least three or four days per week (e.g., every other day), most preferably at least five days per week, six days per week, or seven days per week. The time period of administration can be at least one week, preferably at least one month, more preferably at least two months, most preferably at least three months, for example at least four months. In an embodiment, dosing is at least daily; for example, a subject may receive one or more doses daily. In some embodiments, the administration continues for the remaining life of the individual. In other embodiments, the administration occurs until no detectable symptoms of the condition remain. In specific embodiments, the administration occurs until a detectable improvement of at least one symptom occurs and, in further cases, continues to remain ameliorated.

[0151] The ideal duration of the administration of the composition can be determined by those of skill in the art.

[0152] EXAMPLES

[0153] Example 1

[0154] Investigation of the impact of protein and fiber mixes on the gut microbiota using ProDigest’s short-term single-stage colonic simulation

[0155] Methods and Materials:

[0156] In vitro digestion:

[0157] In vitro digestion simulation model follows the static digestion protocol by international INFOGEST consortia. In short, eight test products (Table 1) were subjected to a full passage through the oral, gastric and small intestinal phases, the latter involving absorption through dialysis. Pre-digestion was considered important as all products contain a fraction of undigestible materials that can be fermented by the gut bacteria. ProDigest further improved this digestion method by incorporating more accurate pH profiles together with a simulation of small intestinal absorption by means of dialysis. The latter enables removal of small molecules from intestinal digests, using 14 kDa dialysis membranes.

[0158] In vitro batch fermentation:

[0159] Fecal material was collected from five healthy adult donors within the framework of a preceding project but only the samples from three donors (A, B and E) were used in the present study. In short, fecal suspensions were prepared and mixed with an internally optimized cryoprotectant. The obtained suspensions were aliquoted, flash frozen and then preserved at - 80°C (cryostock). A short-term screening assay consists of a colonic incubation of a single dose of a test compound under conditions representative for the proximal large intestine, using bacterial inocula from donors. At the start of the short-term colonic incubations, 210 mg FOS was added in the respective reactors (Table 2). Because nutrients of the sugar-depleted nutritional medium will also be fermented by the colonic microbiota, a blank condition containing only the sugar-depleted nutritional medium (without FOS or pre-digested protein solution) was included for each donor. The fecal inocula of donors A, B and E were selected by the customer. All treatment groups and blanks are shown in Table 1 .

[0160] Table 1. Treatment groups

[0161] Addition of prebiotic FOS to protein increased spermidine production by human gut microbiota

[0162] Results of spermidine after 24 hours fermentation are shown in Figure 1 (A). The amount of spermidine was highly variable within each treatment and some treatments (P5 and P7) resulted in a very low amount spermidine close to that of blank control. Despite a higher value seen in some groups (1 , P2, P3, P4, P6 and P8) than blank control, there is no statistical difference in the concentration of spermidine when testing the protein digesta alone. In contrast, addition of prebiotic FOS had significant impacts on spermidine production by human gut microbiota, and algae protein (P8) led to a significant increase of spermidine over blank control as shown in Figure 1 (B).

[0163] The effect of FOS on the production of spermidine (A) was more significant than on putrescine (B) production by the gut microbiota.

[0164] Next, the inventors investigated whether the FOS’s effect was specific to spermidine or also to other polyamines. Shown in Figure 2 (A), all protein groups had a higher amount of spermidine when FOS was added to the fermentation media. Significant differences were found with P5 and P6, and a trend to increase was observed with blank control, P2 and P7.

[0165] In contrast to spermidine, the concentration of putrescine was at least one magnitude higher than spermidine in all conditions as shown in Figure 2 (B). Also, protein digesta and FOS had different effects on putrescine levels compared to spermidine (Figure 2). All groups had detectable amount of putrescine even without FOS. When FOS was added, putrescine concentrations were not changed in all groups.

[0166] FOS rescued the positive relationship between arginine in the proteins and spermidine when fermentation extended to 24 and 48 hours.

[0167] Arginine is the substrate for microbial polyamine production. The inventors next examined whether the amount of arginine in protein sources determines the polyamine concentrations. Shown in Figure 3 to 5, the amount of arginine found in protein sources was positively associated with the amount of spermidine after 6h fermentation (R2 = 0.72, p<0.0001 , Figure 3 (A)), and addition of FOS did not change the relationship (R2 = 0.44, p = 0.0004, Figure 3 (A)). However, when fermentation extended to 24 and 48 hours, no relationship between arginine in the proteins and spermidine can be found in 24h (R2 = 0.1 , p = 0.14, Figure 3 (B)) or 48h (R2 = 0, p = 0.99, Figure 3 (C)). Interestingly, FOS rescued the positive relationship in both 24h and 48h (Figure 3 (B) and Figure 3 (C)), indicating the importance of prebiotics in feeding the gut ecosystem for spermidine production.

[0168] Example 2

[0169] Comparison of proteins of plant and animal origins on the microbial production of polyamines using short-term in vitro fermentation system

[0170] Methods and Materials

[0171] In vitro digestion

[0172] Protein samples and negative control samples were first digested before being fermented by human gut microbiota. The names of the protein samples from plants and animals are listed in Table 2. It is worth noting that organic pea proteins from two different sources (source 1 : Hill pharma incorporate; source 2: Puris) were test and compared. In brief, a three steps protocol mimics the digestion processes in the mouth, stomach and small intestine. In the oral phase, test products were diluted in simulated salivary fluid (1 :1 wt / wt) and normalized to a final concentration of 0.04 g of protein content per mL. CaCI2 (1.5 mM) and amylase (75 U / mL) were added to the mixture and the mixture was incubated at 37oC for 2 min at pH 7.0 under agitation. Since the protein products come with different proportions of protein content, adjustment of weight of protein materials was applied to normalize the protein load at 0.04 g / mL at the oral phase with the exception of faba bean protein concentrate. Actual protein content in faba bean protein concentrate was close to 60% but adjustment was set at 80%, thus less than ideal amount of protein was tested. Then, the oral bolus was mixed simulated gastric fluid at the equal volume (1 :1). In the stomach phase, pepsin (2000 U / mL) and CaCI2 (0.15 mM were added to the mixture before an incubation at 37°C for 2h at pH 3.0. At the end of stomach digestion, gastric chyme was mixed with simulated intestinal fluid at a 1 to 1 ratio. Bile salts (10 mM), CaCI2 (0.6 mM) and pancreatin (trypsin activity at 100 U / mL) were included in the small intestine digestion chyme. Digestion conditions are 2h at 37°C at pH 7.0. Removal of free amino acids and small peptides and characterization of remaining materials

[0173] Removal of small peptides and free amino acids was achieved with methanol precipitation. The samples obtained from in vitro digestion (IVD) were precipitated with 80 % MeOH (1 h at - 20°C) and splitted by centrifugation (13,000 * g at 4°C for 15 min) into a supernatant (S) and a pellet (P). The undigested (P) fraction was lyophilized and stored at -20°C before the fermentation experiment.

[0174] Table 2. Protein samples used in the in vitro digestion and fermentation experiment

[0175] Stool sample collection

[0176] Fecal samples of healthy volunteers were collected under a protocol approved by Lausanne ethical committee (CER-VD) (authorization number: 2020-00304). Inclusion criteria are healthy participant aged from 18-60 years old who provide informed consent and are willing to follow the clinical study protocol. The exclusion criteria are 1) following a particular dietary regime such as vegan, vegetarian, ketogenic or paleo diet; 2) experiencing chronic or recurrent diarrhea with spontaneous bowel movement more than twice a day; 3) antibacterial / antifungal therapy during the 3 months prior to study enrollment; 4) medications or supplements that are known to alter gut function or gut microbiota (i.e. acid antisecretory drugs, pre- / probiotics supplements, laxatives) during the 4 weeks prior to study enrolment, 5) prior gastrointestinal surgery, 6) alcohol intake higher than 2 servings per day; 7) artificially sweetened beverage intake higher than 1000 mL / per day; 8) current or history of gastrointestinal diseases.

[0177] Preparation of stool samples for in vitro fermentation followed the procedure described by Van den Abbeele et al. Briefly, freshly collected stool samples were placed in an air-tight jar equipped with AnaeroGenTM to reduce the exposure of ambient oxygen. Once inside of an anaerobic chamber (Coy Laboratory Products, Grass Lake, Ml, USA), fecal materials were diluted 10 times (w / v) in anaerobic phosphate buffer (0.1 M of NaH2PO4 and 0.1 M of Na2HPO4 in 2:1 ratio) containing 10% glycerol and the aliquots of fecal stocks (25 mL) were stored at -80°C for later use.

[0178] In vitro fermentation

[0179] To prepare a batch fermentation experiment, frozen stool samples were thawed in a water bath at 37°C for 5minutes. Then, samples were centrifugated at 2000Xg for 2 minutes at room temperature, and 250 uL of supernatent was mixed with 10 mL of fermentation media in a sterile Hungate tubes (composition is shown in Table 3). Pellet of MeOH precipitation was resuspended in the fermentation media at the final concentration of 3 mg / mL in a Hungate tube. Fermentation time was performed anaerobically at 37oC for 48h during the screening and 24h in the second round of experiment. Each protein ingredient was tested in three independent fecal samples for allowing statistical analysis.

[0180] Table 3. Composition of fermentation media

[0181] Quantitative analysis of polyamine in the fermentation media

[0182] Quantification of agmatine and other polyamines was documented in an internal analytical report (R0000880953). All samples were prepared according to the analytical approach developed in-house as reported in the R&D Memo: “Polyamines in bacterial media - Analytical approach using LC-HRMS”. For details of analytical approach and method validation, please refer to the note and document mentioned above.

[0183] Among test proteins, faba bean protein resulted in the highest amount of spermidine followed by pea where other proteins did not result in different amount of spermidine from control, and test proteins had no effect on amounts of putrescine.

[0184] Spermidine and putrescine concentrations at 24h were shown in Figure 4 (A) and Figure 4 (B), respectively. Among all test proteins, faba bean protein resulted in the highest amount of spermidine followed by pea 870 where other proteins did not result in different amount of spermidine from control (Enzyme only) (Figure 4 (A)). Contrary to spermidine, the amounts of putrescine were similar in all groups (Figure 4 (B)).

[0185] Starch and fiber are expected to be found in the non-protein fraction. As the inventors observed in Example 1 that FOS enhanced spermidine production, it is logical to expect starch or fibers in faba bean contrate also promotes the spermidine synthesis just like FOS.

[0186] The inventors found that fiber (FOS) and less refine protein source, faba bean protein concentrate are good nutritional candidates to enhance the production of anti-ageing molecule spermidine.

[0187] The inventors also expect other protein-prebiotic combinations or less refined protein preparations from soy, pea, lentil, or other plant sources would have similar advantage over animal proteins for spermidine production.

[0188] The studies demonstrates that intake of a composition of a protein source and a prebiotic source increases spermidine production by the gut microbiota.

[0189] It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present subject matter and without diminishing its intended advantages. It is therefore intended that such changes and modifications be covered by the appended claims.

Claims

CLAIMS1 . A composition comprising at least one source of protein and at least one source of prebiotic for use in increasing spermidine production by the gut microbiota of a subject.

2. The composition as claimed in claim 1 , wherein the source of protein is at least one selected from the group: pulses, grain, nuts, algae, oil crop and / or its press cakes, animal proteins or combination thereof.

3. The composition as claimed in claim 2, wherein pulses are at least one selected from the group comprising faba bean, chick pea, green pea, lentil.

4. The composition as claimed in claim 2, wherein grains are at least one selected from the group comprising rice, oat, quinoa, wheat, barley.

5. The composition as claimed in claim 2, wherein nuts are at least one selected from walnut, almond, pistachio.

6. The composition as claimed in claim 2, wherein algae are at least one selected from seaweed, microalgae.

7. The composition as claimed in claim 2, wherein oil crop and / or its press cakes are selected from soy, canola, peanut, bambara.

8. The composition as claimed in claim 2, wherein animal proteins are at least one selected from milk, egg, seafood, poultry, red meat, white meat.

9. The composition as claimed in any of the preceding claims, wherein the source of prebiotic is at least one oligosaccharide.

10. The composition as claimed in claim 9, wherein the at least one oligosaccharide is selected from the group: fructo-oligosaccharide, inulin, guar gum, galactooligosaccharide, betaglucan, resistant starch, arabinoxylan, pectin, raffinose, polydextrose, agar, alginate or combination thereof.11 . The composition as clamed in any of the preceding claims, comprising the at least one source of protein and the at least one source of prebiotic in an effective amount.

12. The composition according to any of the preceding claims wherein the source of protein is administered to the subject in an amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.

13. The composition according to any of the preceding claims wherein the source of prebiotic is administered to the subject in an amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

14. The composition according to any of the preceding claims, wherein the at least one source of protein is faba bean and the at least one source of prebiotic is fructo-oligosaccharide.

15. The composition according to claim 8,14wherein it comprises:- faba bean protein in the amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.- fructo-oligosaccharide in the amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

16. The composition according to any of claims 1 to 13, wherein the at least one source of protein is pulses and the at least one source of prebiotic is fructo-oligosaccharide.

17. The composition according to claim 10, wherein it comprises:- pulses protein in the amount of from at least about 1 mg to at least about 2400 mg per kg of a body weight of the subject.- fructo-oligosaccharide in the amount of from at least about 10 mg to at least about 1000 mg per kg of a body weight of the subject.

18. The composition as claimed in any one of the preceding claims for use in preventing and / or treating age-related disorders.

19. The composition as claimed in claim 18, wherein age-related disorder is at least one selected from the group: a physiological state or disorder related to cell ageing, a physiological state linked to metabolic fatigue in one or more cells, excessive catabolism, oxidative stress, reduced antioxidant capacity and / or mitochondrial function, impaired mobility, healthspan and / or lifespan in a subject, muscle dysfunction, including sarcopenia, frailty, inclusion body myositis, myopathy / myolysis induced by drugs such as corticosteroids or statins, muscle wasting induced by immobilization or hospitalization, muscle weakness, immune function disorders, cognitive function disorders, including delirium, dementia, learning disorder, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD), cardiovascular disease, compromised intestine barrier integrity or leaky gut, compromised gut microbiome, age- associated inflammation.

20. The composition as claimed in any one of the preceding claims for use in: increasing (i) resistance to age-related pathologies, (ii) improving a physiological state or disorder related to cell ageing, (iii) improving a physiological state linked to metabolic fatigue in one or more cells, (iv) increasing mitochondrial energy in one or more cells, (v) increasing antioxidant capacity, reducing oxidative stress and / or enhancing mitochondrial function, (vi) improving mobility and / or (vii) improving healthspan and / or lifespan in a subject.

21. The composition according to any of the preceding claims, wherein the composition is in the form of food composition, dietary supplement, nutritional composition, nutraceutical, powdered nutritional product to be reconstituted in water or milk before consumption, food additive, medicament, beverage and drink, including coffee-based product, oral nutritional supplement (ONS), a complete nutritional formula, a pharmaceutical, a medical or a food product.

22. The composition as claimed in any of the preceding claims, wherein the subject is a human, a farm animal or a pet.

23. Use of the composition of any one of the preceding claims in the subject in need of at least one increasing (i) resistance to age-related pathologies, (ii) improving a physiological state or disorder related to cell ageing, (iii) improving a physiological state linked to metabolic fatigue in one or more cells, (iv) increasing mitochondrial energy in one or more cells, (v) increasing antioxidant capacity, reducing oxidative stress and / or enhancing mitochondrial function, (vi) improving mobility and / or (vii) improving healthspan and / or lifespan in an individual.

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