Compositions and methods using spermidine to reduce biological age of an adult animal

Abstract

Compositions and methods reduce the biological age and / or reduce the rate of increase of the biological age of an adult animal. More specifically, the present disclosure relates to administering a formulation comprising an effective amount of spermidine, alone or in combination with a dietary regime such as a standard diet, caloric restriction, senior diet or high protein diet. The formulation can concomitantly reduce the biological age of a tissue, organ or physiological system in the adult animal in need; or treat, prevent, or delay the onset of an organ disease, or decline of organ function, or condition relating thereto in an adult animal in need.

Description

TITLECOMPOSITIONS AND METHODS USING SPERMIDINE TO REDUCE BIOLOGICAL AGE OF AN ADULT ANIMALTECHNICAL FIELD

[0001] The present disclosure generally relates to compositions and methods to reduce the biological age and / or reduce the rate of increase of the biological age of an adult animal. More specifically, the present disclosure relates to administering a formulation comprising an effective amount of spermidine, alone or in combination with a dietary regime such as a diet in which protein is 18% to 60% of protein / dry matter (w / w) for dogs and 26% to 60% of protein / dry matter (w / w) for cats. The formulation can concomitantly reduce the biological age of a tissue, organ or physiological system in the adult animal in need; or treat, prevent, or delay the onset of an organ disease, or decline of organ function, or condition relating thereto in an adult animal in need.BACKGROUND

[0002] Chronological age is known to be a major indicator of general health status, with increasing chronological age associated with reduced health. However, depending on genetics, nutrition, and lifestyles, individuals may age slower or faster than their chronological age.

[0003] Chronological age may therefore not always reflect an individual’s rate of aging or risk of reduced health. On the other hand, the biological age of an individual (based on e.g. clinical biochemistry and cell biology measures) can vary compared to others of the same chronological age.

[0004] Highly digestible and high-quality protein diets are generally recommended based upon the chronological age of a dog. For example, it may be recommended that a dog is switched to a senior diet around 7 or 8 years old. However, the determination of an increased biological age for a dog compared to its chronological age may allow a determination to provide additional nutritional solution to the animal.

[0005] There is thus a need for methods and compositions effective in reducing biological age and conditions related thereto, of an adult animal having an increased biological age compared to its chronological age.SUMMARY

[0006] Accordingly, in a general embodiment, the present disclosure provides a method for reducing the biological age and / or for reducing the rate of increase of the biological age of an adult animal in need, the method comprising: feeding the adult animal a composition comprising an effective amount of spermidine. As detailed in the experimental data set forth later herein, the inventors surprisingly found that spermidine induces reduction in biological age. This reduction in biological age occurred regardless of the diet the spermidine was administered in combination with.

[0007] The present disclosure also provides a method for reducing the biological age of a tissue, organ or physiological system in an adult animal in need, the method comprising feeding the adult animal a composition comprising an effective amount of spermidine.

[0008] The present disclosure further provides a method for treating, preventing, or delaying the onset of an organ disease, or decline of organ function, or condition relating thereto in an adult animal in need, the method comprising feeding the adult animal a composition comprising an effective amount of spermidine.

[0009] Suitably, the present methods may comprise selecting and / or applying the composition comprising an effective amount of spermidine to an animal following a determination that the animal’s biological age is greater than its chronological age. The increased biological age may be in a tissue, in an organ or in a physiological system such that immune, gastrointestinal, urinary, muscular, cardiovascular, and / or neurological system.

[0010] Suitably, improving the biological age of an animal may refer to a reduction in the difference between the biological age and chronological age of the animal, where the biological age of the animal is greater than its chronological age. Alternatively, a worsening in the biological age of an animal may refer to an increase in the difference between the biological age and chronological age of the animal, where the biological age of the animal is greater than its chronological age.

[0011] Suitably, improving the biological age of an animal may refer to a reduction in the rate of change between the biological age and chronological age of the animal, where the biological age of the animal is greater than its chronological age. For example, an animal’s biological age may have been increasing by 1.5 years per 1 year increase in chronological age.

[0012] Suitably, whilst the present composition may be effective for animals based on chronological age, it may be particularly effective when applied to an animal with an increased biological age compared to its chronological age. As such, the present method mayadvantageously be beneficial for an animal that has an increased likelihood to respond, or improved magnitude of response, to the present composition.

[0013] The composition comprising spermidine may be part of or administered together with, a dietary regime. The dietary regime may be a diet in which protein is 18% to 60% of protein / dry matter (w / w) for dogs and 26% to 60% of protein / dry matter (w / w) for cats.

[0014] An advantage of one or more embodiments provided by the present disclosure is to improve the condition of animals, ageing animals, or ageing humans.

[0015] Still another advantage of one or more embodiments provided by the present disclosure is to increase the survivability of an ageing individual.

[0016] An additional advantage of one or more embodiments provided by the present disclosure is to increase mobility in the animal.

[0017] Additional features and advantages are described in, and will be apparent from, the following Detailed Description and the Figures.BRIEF DESCRIPTION OF THE DRAWINGS

[0018] FIG. 1 shows the amounts of protein, carbohydrates, and fats of the first diet fed to the mice versus the second diet used in the examples.

[0019] FIG. 2 shows the effects on biological age of administering a composition including an effective amount of spermidine in a first diet and a second diet, respectively. Biological age has been estimated by analysis of DNA methylation using the intervention muscle clock.DETAILED DESCRIPTION

[0020] Definitions

[0021] 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.

[0022] All percentages expressed herein are by weight of the total weight of the composition unless expressed otherwise. As used herein, “about,” “approximately” and “substantially” are understood to refer to numbers in a range of numerals, for example the range of -10% to +10% of the referenced number, preferably -5% to +5% of the referenced number, more preferably -1% to +1% of the referenced number, most preferably -0.1% to +0.1% of the referenced number. All numerical ranges herein should be understood to include all integers, whole or fractions, within the range. Moreover, these numerical ranges should be construedas providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.

[0023] 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. For example, reference to “an ingredient” or “a method” includes a plurality of such “ingredients” or “methods.”

[0024] 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 and methods 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 or steps identified.

[0025] The terms “at least one of’ and “and / or” used respectively in the context of “at least one of X and Y” and “X and / or Y” should be interpreted as “X without Y,” or “Y without X,” or “both X and Y.” 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.

[0026] As used herein, “related to,” “associated with” and “linked with” mean occurring concurrently, preferably mean caused by the same underlying condition, more preferably mean that one of the identified conditions is at least indirectly caused by the other identified condition, and most preferably mean that one of the identified conditions is directly caused by the other identified condition.

[0027] The terms “food,” “food supplement,” “food product” and “food composition” mean a product or composition that is intended for ingestion by an individual such as a human and provides at least one nutrient to the individual. The compositions of the present disclosure, including the many embodiments described herein, can comprise, consist of, or consist essentially of the elements disclosed herein, as well as any additional or optional ingredients, components, or elements described herein or otherwise useful in a diet supplement.

[0028] “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, includingtherapeutic 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. 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 nonlimiting examples, a treatment can be performed by a patient, a caregiver, a doctor, a nurse, or another healthcare professional.

[0029] The relative terms “reduce” or “reducing” and the like refer to the effects of the composition comprising a spermidine (disclosed herein) relative to a composition without spermidine or with decreased levels of spermidine but otherwise identical.

[0030] 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.

[0031] “Animal” includes, but is not limited to, mammals, which includes but is not limited to rodents, aquatic mammals, domestic animals such as dogs and cats, farm animals such as sheep, pigs, cows and horses, and humans.

[0032] The term “elderly” in the context of a human means an age from birth of at least 55 years, preferably above 63 years, more preferably above 65 years, and most preferably above 70 years. The term “older adult” or “ageing individual” in the context of a human means an age from birth of at least 45 years, preferably above 50 years, more preferably above 55 years, and includes elderly individuals.

[0033] For other animals, an “older adult” or “ageing individual” has exceeded 50% of the average lifespan for its particular species and / or breed within a species. An animal is considered “elderly” if it has surpassed 66% of the average expected lifespan, preferably if it has surpassed the 75% of the average expected lifespan, more preferably if it has surpassed 80% of the average expected lifespan. An ageing cat or dog has an age from birth of at least about 5 years. An elderly cat or dog has an age from birth of at least about 7 years.

[0034] Chronological age may be defined as the amount of time that has passed from the subject’s birth to the given date. Chronological age may be expressed in terms of years,months, days, etc. In certain embodiments, the animal is a dog and may be at least about 2 years old. Suitably, the dog may be at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9 or at least about 10 years old.

[0035] A biological age for a particular tissue, organ, or physiological system may be determined using a DNA methylation profile comprising, or consisting of, methylation sites from genes that are preferentially or specifically expressed by that tissue, organ, or physiological system. Classifications of genes by a particular tissue, organ, or physiological system are publically available at, for example, Gene Ontology (http: / / geneontology.org / ), the KEGG pathway database (https: / / www.genome.jp / kegg / ), or MSIgDB (https: / / www.gsea- msigdb.org / gsea / msigdb / index.jsp).

[0036] The term “composition” may mean a food, beverage, dietary supplement, complete nutrition or oral nutritional supplement (ONS) or medical food composition, or mixture thereof.

[0037] As used herein, “complete nutrition” contains sufficient types and levels of macronutrients (protein, fats and carbohydrates) and micronutrients to be sufficient to be a sole source of nutrition for the animal to which the composition is administered. Individuals can receive 100% of their nutritional requirements from such complete nutritional compositions.

[0038] 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 an 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.

[0039] Embodiments

[0040] An aspect of the present disclosure is a method for reducing the biological age and / or reducing the rate of increase of the biological age of an adult animal in need. The method comprises feeding the adult animal a composition comprising an effective amount of spermidine. The present methods may comprise determining a DNA methylation profile to determine a biological age of an animal, e.g. a human or a dog using any suitable method.

[0041] The composition comprising an effective amount of spermidine and / or functional derivatives, can be administered to an animal such as a human, e.g., an ageingindividual or a pet animal, in a therapeutically effective dose. The therapeutically effective dose can be determined by the person skilled in the art and will depend on a number of factors known to those of skill in the art, such as the severity of the condition and the weight and general state of the individual.

[0042] Non-limiting examples of suitable forms of spermidine include spermidine trihydrochloride, spermidine phosphate hexahydrate, spermidine phosphate hexahydrate, and L-arginyl-3,4-spermidine. Wheat germ is rich in spermidine. Therefore, some embodiments of the composition comprise wheat germ and / or enriched wheat germ extracts that provide at least a portion of the spermidine in the composition.

[0043] In particular embodiments, the method is a treatment that augments the plasma spermidine level in an individual, for example to a level in the range of 50 to 6000 nmol / L plasma, preferably 100 to 6000 nmol / L plasma. The method can comprise administering daily the spermidine in the weight range of 0.05 mg - 1 g per kg body weight, preferably 1 mg -200 mg per kg body weight, more preferably 5 mg - 150 mg per kg body weight, even more preferably 10 mg - 120 mg per kg body weight, or most preferably 40 mg - 80 mg per kg body weight. In some embodiments, the method can comprise administering 10 mg - 100 mg of spermidine per serving, for example about 16 mg per serving of a food item or about 80 mg per serving of a supplement or treat.

[0044] The composition is preferably administered to the individual at least two days per week, more preferably at least three days per week, most preferably all seven days of the week; for at least one week, at least one month, at least two months, at least three months, at least six months, or even longer. In some embodiments, the composition is administered to the individual consecutively for a number of days, for example at least until a therapeutic effect is achieved. In an embodiment, the composition can be administered to the individual daily for at least 30, 60 or 90 consecutive days.

[0045] The above examples of administration do not require continuous daily administration with no interruptions. Instead, there may be some short breaks in the administration, such as a break of two to four days during the period of administration. The ideal duration of the administration of the composition can be determined by those of skill in the art.

[0046] In a preferred embodiment, the composition is administered to the individual orally or enterally (e.g. tube feeding). For example, the composition can be administered to the individual as a beverage, a capsule, a tablet, a powder or a suspension.

[0047] The composition can be any kind of composition that is suitable for human and / or animal consumption. For example, the composition may be selected from the group consisting of food compositions, dietary supplements, nutritional compositions, nutraceuticals, powdered nutritional products to be reconstituted in water or milk before consumption, food additives, medicaments, beverages and drinks. In an embodiment, the composition is an oral nutritional supplement (ONS), a complete nutritional formula, a pharmaceutical, a medical or a food product. In a preferred embodiment, the composition is administered to the individual as a beverage. The composition may be stored in a sachet as a powder and then suspended in a liquid such as water for use.

[0048] In an embodiment, the composition includes a source of protein. The composition can comprise a mixture of one or more types of protein, for example one or more (i) peptides, (ii) longer chains of amino acids, or (iii) free form amino acids; and the mixture is preferably formulated to achieve a desired amino acid profile / content.

[0049] At least a portion of the protein can be from animal or plant origin, for example dairy protein such as one or more of milk protein, 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. Additionally or alternatively, at least a portion of the protein can be plant protein such as one or more of soy protein or pea protein.

[0050] In an embodiment, the composition includes a source of carbohydrates. Any suitable carbohydrate may be used in the composition including, but not limited to, starch (e.g., modified starch, amylose starch, tapioca starch, com starch), sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrin, xylitol, sorbitol or combinations thereof.

[0051] The source of carbohydrates is preferably not greater than 50 energy % of the composition, more preferably not greater than 36 energy % of the composition, and most preferably not greater than 30 energy % of the composition. The composition can have a high proteimcarbohydrate energy ratio, for example greater than 0.66, preferably greater than 0.9 and more preferably greater than 1.2.

[0052] In an embodiment, the composition includes a source of fat. The source of fat may include any suitable fat or fat mixture. Non-limiting examples of suitable fat sources include vegetable fat, such as olive oil, corn oil, sunflower oil, high-oleic sunflower, rapeseed oil, canola oil, hazelnut oil, soy oil, palm oil, coconut oil, blackcurrant seed oil, borage oil, lecithins, and the like, animal fats such as milk fat; or combinations thereof. In some embodiments, the source of fat is less than 20% of the composition.

[0053] In a preferred embodiment, the composition further contains an autophagy inducer selected from the group consisting of urolithin (e.g., Urolithin A, B or D), rapamycin, Torinl, valproic acid, polyphenols (e.g., resveratrol), caffeine, metformin, 5' AMP-activated protein kinase (AMPK) activators, L-type calcium channel inhibitors, and mixtures thereof. Non-limiting examples of suitable autophagy inducers are palmitic acid, 5-aminoimidazole-4- carboxamide riboside (AICAR), verapamil, nifedipine, diltiazem, piperazine phenothiazine derivatives (e.g., trifluoperazine), ketones (e.g., beta-hydroxybutyrate, ketone salts, or ketone ester derivatives) and mixtures thereof.

[0054] In some embodiments, the composition is administered to the individual in a single dosage form, i.e. all compounds are present in one product to be given to an individual in combination with a dietary regime.

[0055] The dietary regime may be a meal, a regime of meals, a supplement or a regime of supplements, or combinations of a meal and a supplement, or combinations of a meal and multiple supplements.

[0056] The dietary intervention or dietary product described herein may be any suitable dietary regime, for example, a standard diet, a calorie-restricted diet, a senior diet, a high protein diet, a phosphorous diet, potassium supplement diet, polyunsaturated fatty acids (PUFA) supplement diet, anti-oxidant supplement diet, a vitamin B supplement diet, liquid diet, selenium supplement diet, or diets supplemented with carnitine, branched chain amino acids or derivatives, or any combination of the above.

[0057] A dietary intervention may be determined based on the baseline maintenance energy requirement (MER) of the dog. Suitably, the MER may be the amount of food that stabilizes the dog’s body weight (less than 5% change over three weeks).

[0058] Suitably, a calorie-restricted diet may comprise about 50%, about 55%, about 60%, about 65%, about 75%, about 80%, about 85%, or about 90% of the dog’s MER. Suitably, a calorie-restricted diet may comprise about 60% or about 75% of the dog’s MER.

[0059] In an embodiment, the composition has a protein / energy ratio greater than 6 g protein / 100 kcal, preferably greater than 9 g protein / 100 kcal. In an embodiment, the protein is between 18% and 60% (w / w) of the composition on a dry matter basis.

[0060] The dietary regime may comprise a food, supplement and / or drink that comprises a nutrient and / or bioactive that mimics the benefits of caloric restriction (CR) without limiting daily caloric intake. For example, the food, supplement and / or drink may comprise a functional ingredient(s) having CR-like benefits. Suitably, the food, supplement and / or drink may comprise an autophagy inducer. Suitably, the food, supplement and / or drinkmay comprise fruit and / or nuts (or extracts thereof). Suitable examples include, but are not limited to, pomegranate, strawberries, blackberries, camu-camu, walnuts, chestnuts, pistachios, pecans. Suitably, the food, supplement and / or drink may comprise probiotics with or without fruit extracts or nut extracts.

[0061] The methods described herein can be useful for i) reducing the biological age and / or reducing the rate of increase of the biological age of an animal or ii) reducing the biological age of a tissue, organ or physiological system as well as iii) treating, preventing, or delaying onset of an organ disease or organ functional decline, or conditions relating thereto for any animal.

[0062] The individual in need thereof may be an elderly animal or an elderly human. The individual in need thereof may be an ageing human or an ageing animal. The individual in need thereof may have a biological age that is greater than a chronological age. Additionally, the individual in need thereof may have a rate of change between the biological age that is greater than a rate of change to the chronological age.

[0063] In an embodiment, the increased biological age may be in a tissue, in an organ or in a physiological system such that immune, gastrointestinal, urinary, muscular, cardiovascular, and / or neurological system. In a preferred embodiment, the tissue is liver, kidney or skeletal muscle. Non-limiting examples of such muscle include one or more of the following: vastus lateralis, gastrocnemius, tibialis, soleus, extensor, digitorum longus (EDL), biceps femoris, semitendinosus, semimembranosus, gluteus maximus, extra-ocular muscles, face muscles or diaphragm.

[0064] In an embodiment, the animal in need has a deficiency in spermidine in one or more of his tissue.

[0065] Those skilled in the art will understand that they can freely combine all features of the present invention described herein, without departing from the scope of the invention as disclosed.EXAMPLES

[0066] The following non-limiting example generally illustrates the concepts underlying the embodiments disclosed herein.

[0067] The invention will now be further described by way of examples, which are meant to serve to assist the skilled person in carrying out the invention and are not intended in any way to limit the scope of the invention.

[0068] Example 1

[0069] Material and methods

[0070] Animal care

[0071] All animal experiments comply with the International Guiding Principles for Biomedical Research Involving Animals developed by the Council for International Organizations of Medical Sciences and the following guidelines for animal welfare. Mice aged 18 months were kept in a temperature- and humidity-controlled environment with a 12: 12- h light-dark cycle. Mice had access to nesting and enrichment materials and were provided with ad libitum access to water and food.

[0072] Mice treatment and sample collection

[0073] In the experiment, mice were fed either (i) a first diet, (ii) the first diet supplemented with spermidine, (iii) a second diet, or (iv) the second diet supplemented with spermidine. Figure 1 shows the differences in the first diet versus the second diet fed to the mice. The first diet was composed of 16% fats, 20% proteins, and 64% carbohydrates. The second diet was composed of 16% fats, 60% proteins, and 24% carbohydrates. These amounts are percentages of total energy of the feed. Animals found dead or euthanized at the humane endpoint for pain or physical distress, have been excluded from the analysis. At the end of the treatment, mice were euthanized with isoflurane after 2-hours fasting. Muscle gastrocnemius has been dissected samples and snap frozen in liquid nitrogen and stored at -80°C.

[0074] DNA extraction

[0075] DNA extraction was performed on 100 mg of muscle using QIAGEN DNeasy Blood and Tissue kit (Cat. No: 69506) and following manufacturer recommendations. DNA was quantified using the DropSense technology (from Unchained Labs).

[0076] Methylation detection

[0077] Bisulfite conversion was performed on 500ng of DNA using the Zymo EZDNA methylation kit (Cat. No: D5001) and following manufacturer recommendations. 200ng of converted DNA was used to proceed to the Infmium HD methylation assay protocol following manufacturer recommendations. DNA methylation data were generated using custom arrays: HorvathMammalMethyl40 (Cat. No: 20027399) from Illumina with the consent of the Clock Foundation (Torrance, California), who developed the arrays. Arrays were scanned using the iScan from Illumina.

[0078] Data analysis

[0079] To calculate biological age, raw data have been analysed using algorithms (clocks) developed by the Clock Foundation. An intervention muscle clock, trained on tissuespecific data from mice that underwent caloric restriction, was used. This clocks is based on methylation analysis of different CpGs and show robust chronological age predicting power. The results obtained for each sample corresponds to the biological age in years. Data were plotted and analysed using GraphPad Prism 9. Ordinary One-way ANOVA followed by Dunnett’s multiple comparisons was performed to test the effect of age and spermidine supplementation compared to their respective controls.

[0080] Results

[0081] The results show that biological age decreased in mice fed the first diet with spermidine compared to the mice administered the first diet untreated. Additionally, the biological age decreased in mice fed the second diet with spermidine compared to mice fed the first diet with spermidine. Surprisingly, daily administration of spermidine at the dose of 17.5 mg / kg / day for 12 weeks significantly reduced biological age in mice fed both the first diet and the second diet. As shown in Figure 2, treatment with spermidine led to a reduction of muscle biological age measured with the interventions muscle clock.

Claims

CLAIMS1. A method for reducing the biological age and / or for reducing the rate of increase of the biological age of an adult animal in need, the method comprising: feeding the adult animal a composition comprising an effective amount of spermidine.

2. A method for reducing the biological age of a tissue, organ or physiological system in an adult animal in need, the method comprising feeding the adult animal a composition comprising an effective amount of spermidine.

3. A method according to claim 2, wherein the tissue, organ or physiological system is immune, gastrointestinal, urinary, muscular, cardiovascular, and / or neurological system.

4. A method according to claim 2, wherein the tissue has a deficiency in spermidine.

5. A method according to any of claims 2-4, wherein the tissue is skeletal muscle.

6. A method for treating, preventing, or delaying the onset of an organ disease, or decline of organ function, or condition relating thereto in an adult animal in need, the method comprising feeding the adult animal a composition comprising an effective amount of spermidine.

7. A method according to any preceding claim, wherein the animal in need has an increased biological age compared to its chronological age.

8. A method according to any preceding claim, wherein the increased biological age is determined from the DNA methylation profile of the adult animal.

9. A method according to any preceding claim, wherein the animal is an ageing animal.

10. A method according to any preceding claim, wherein the animal is a pet animal.

11. A method according to any preceding claim, wherein the animal is a human.

12. A method according to any preceding claim, wherein the composition is selected from the group consisting of food compositions, dietary supplements, nutritional compositions, nutraceuticals, powdered nutritional products to be reconstituted in water or milk before consumption, food additives, medicaments, drinks, dry or wet pet food and combinations thereof.

13. A method according to any preceding claim, wherein the composition is part of, or administered together with, a dietary regime.

14. A method according to claim 13, where the dietary regime comprises a diet in which protein is 18% to 60% of protein / dry matter (w / w) for dogs.

15. A method according to claim 13, where the dietary regime comprises a diet in which protein is 26% to 60% of protein / dry matter (w / w) for cats.

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