GB2628186A

GB2628186A - Gender-specific compositions

Info

Publication number: GB2628186A
Application number: GB2303965.4A
Authority: GB
Inventors: De Jong Jelle; Gross Gabriele; Hendriks Lauren; Keijer Jaap; Gijsbert Nieuwenhuizen Arie; Van Den Hoek Anita; Alida Van Diepen Janna; Verschuren Lars
Original assignee: Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO; Wageningen Universiteit; MJN US Holdings LLC
Current assignee: Nederlandse Organisatie voor Toegepast Natuurwetenschappelijk Onderzoek TNO; Wageningen Universiteit; MJN US Holdings LLC
Priority date: 2023-03-17
Filing date: 2023-03-17
Publication date: 2024-09-18
Also published as: WO2024194258A1

Abstract

A composition for use in the treatment, reduction and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises at least one of: docosahexaenoic acid (DHA), eicosapenaenoic acid (EPA), resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, arachidonic acid (ARA), krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof. The age-related loss of muscle mass and/or muscle function may comprise sarcopenia. The composition may be a synthetic composition, and may be a supplement, a nutritional composition, or combination thereof. The female subject may be a middle-aged or elderly adult. The composition may be provided in a single dosage. Also claimed is a composition for use as defined above, comprising at least one of the components listed above or any combination thereof, and at least one of: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof. Also claimed is a composition comprising at least two of the components listed above or any combination thereof.

Description

Gender-Specific Compositions

Field of the Invention

[0001] The present application relates to compositions for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function. In particular, compositions for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject.

Background

[0002] Sarcopenia is a form of age-related loss of muscle mass and/or muscle function. Sarcopenia is the involuntary, progressive loss of muscle mass, function, and strength during ageing, which contributes to decreased quality of life and increased risk for disability, morbidity, and even mortality (Calvani et al., J. Frailty Aging, 2013; Pascual-Fernandez et al., Int. J. Mol. Sci., 2020). It currently affects approximately 6 to 22% of adults over the age of 65 (Dent et al., J. Nutr. Health Aging, 2018). Although the rate of muscle mass loss varies among different studies, it has been estimated to occur from as early as 35 years old, at a rate of 0.8% per year, increasing to a rate of 3% per year by 60 years old. Generally, this loss of muscle mass and strength is caused by muscle atrophy, as well as muscle cell death, and is greater in sedentary individuals than active individuals. Simultaneously, fat mass can increase with approximately 0.45 kilograms per year (kg/year) in an average adult between 30 and 60 years of age (Calvani et al., J. Frailty Aging, 2013). This masks the change in body composition and leads to sarcopenic obesity, which further increases the risk of disabilities. The onset of sarcopenia has been attributed to a combination of several factors, such as loss of motor neurons, endocrine hormonal alterations, increased production of pro-inflammatory cytokines, impaired satellite cell function, loss of muscle motor units, and nutritional and lifestyle changes. Collectively, these factors cause an imbalance between anabolic and catabolic processes, leading to degradation of muscle protein, loss of muscle fibres, insufficient replacement of satellite cells, and thus, eventually loss of muscle mass. However, the molecular mechanisms and pathways underlying the pathogenesis of sarcopenia still remain unknown (Ziaaldini et aL, Front. Med., 2017).

[0003] An effective treatment for sarcopenia is yet to be established (Calvani et at, J. Frailty Aging, 2013). The current recommendation for adults with sarcopenia is to combine nutritional protein supplementation (1 to 1.2 g/kg body weight per day) with physical exercise (Bauer et al., J. Am. Med. Dir. Assoc., 2013; Dent et al., J. Nutr. Health Aging, 2018). However, protein supplementation is not an effective intervention for all subjects and undertaking physical exercise may be difficult for fragile and/or infirmed elderly subjects, who lack sufficient mobility or motivation to regularly adhere to the prescribed exercise plan.

[0004] There have been various efforts to develop pharmacological and/or nutritional interventions for sarcopenia, in conjunction with and without exercise (Ganapathy & Nieves, Nutrients, 2020). Lo et a/. compiled a literature study of 34 clinical trials that involved nutritional supplements, protein supplements, and/or exercise interventions for sarcopenia, in elderly populations (J. Orthop. Translat., 2020). The results showed that only studies involving some form of physical activity as an intervention showed significant improvements in muscle strength and performance. Thus, it is clear that the nutritional sarcopenia interventions trialled to-date have not been effective, nor did they exhibit any synergistic effects when combined with exercise.

[0005] Accordingly, there exists a need for novel, safe, and effective interventions for the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function. It would be preferable if the novel interventions were nutritional interventions i.e. they involved nutritional compositions. It would be desirable if the nutritional compositions exhibited an effect in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in absence of exercise, and exhibited a synergistic effect in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function when combined with exercise.

Summary of Invention

[0006] In a first aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises at least one of the following: docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, arachidonic acid (ARA), krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof.

[0007] In a second aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises DHA and/or EPA in an amount of 100 milligrams (mg) to 2500 mg.

[0008] In a third aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises resveratrol in an amount of 5 mg to 750 mg.

[0009] In a fourth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises milk fat globule membrane (MFGM) in an amount of 25 mg to 1000 mg.

[0010] In a fifth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises vitamin D in an amount of 5 micrograms (pg) to pg.

[0011] In a sixth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises folate or folic acid in an amount of 50 pg to 1500 pg.

[0012] In a seventh aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female 30 subject, wherein the composition comprises ARA in an amount of 200 mg to 1000 mg.

[0013] In an eighth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises krill oil in an amount of 100 mg to 1500 mg.

[0014] In a ninth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises quercetin, green tea extract, and/or grape seed extract in an amount of 25 mg to 1500 mg.

[0015] In a tenth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises curcumin in an amount of 10 mg to 3000 mg.

[0016] In an eleventh aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises vitamin D and at least one of folate, folic acid, or a combination thereof.

[0017] In a twelfth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises DHA and/or EPA, vitamin D and at least one of folate, folic acid, or a combination thereof [0018] In a thirteenth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises DHA and/or EPA, resveratrol, MFGM, vitamin D and at least one of folate, folic acid, or a combination thereof.

[0019] Preferably, the age-related loss of muscle mass and/or muscle function comprises sarcopenia.

[0020] Preferably, the composition is a supplement, a nutritional composition, or a combination thereof [0021] Preferably, the composition is a synthetic composition.

[0022] Preferably, the female subject is a middle-aged adult or an elderly adult.

15 20 25 [0023] Preferably, the daily dosage of the composition is provided in a single dosage.

[0024] In a fourteenth aspect, there is provided a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject comprising: * at least one of the following: DHA, EPA, resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, ARA, krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof; and, * at least one of the following: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof.

[0025] In a fifteenth aspect, there is provided a composition comprising at least two of the following: * at least one of DHA, EPA, ARA, or any combination thereof; * resveratrol; * quercetin, green tea extract, and/or grape seed extract; * curcumin; * MFGM and/or krill oil; * vitamin D; and, * folate and/or folic acid. 25 [0026] In a sixteenth aspect, there is provided a composition comprising: * at least two of the following: o at least one of DHA, EPA, ARA, or any combination thereof; o resveratrol; o quercetin, green tea extract, and/or grape seed extract; o curcumin; o MFGM and/or krill oil; o vitamin D; and, o folate and/or folic acid; and, * at least one of the following: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof [0027] In a seventeenth aspect, there is provided the composition of the sixteenth aspect for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject. Preferably, the age-related loss of muscle mass and/or muscle function comprises sarcopenia.

Definitions [0028] "Milk' means a substance that has been drawn or extracted from the mammary gland of a mammal, or any non-dairy, vegan-friendly, and/or plant-based alternative, such as coconut milk, soy milk, almond milk, oat milk, hazelnut milk, pea milk, potato milk, quinoa milk, an animal-free milk, or any combination thereof. The "milk' may be a fermented (or cultured) milk i.e. where the milk has been fermented with one or more probiotics e.g. Lactobacilli, Bifidobacteria, etc. [0029] "Nutritional composition" means a substance or composition that satisfies at least a portion of a subject's nutrient requirements. "Nutritional composition(s)" may refer to liquids, powders, solutions, gels, pastes, solids, concentrates, suspensions, ready-to-use forms of enteral formulas, and/or oral formulas.

[0030] "Reconstituted solution", in terms of the present disclosure, means the solution prepared when a diluent (e.g. water, saline, milk, etc.) is added to an ingredient (e.g. a powder, a solution, a gel, a suspension, a paste, a solid, a liquid, a liquid concentrate, etc.).

[0031] The term "synthetic" when applied to a composition, a nutritional composition, or a mixture means a composition, nutritional composition, or mixture obtained by biological and/or chemical means, which can be chemically identical to the mixture naturally occurring in mammalian milks. A composition, nutritional composition, or mixture is said to be "synthetic" if at least one of its components is obtained by biological (e.g. enzymatic) and/or chemical means.

[0032] The term "degree of hydrolysis" refers to the extent to which peptide bonds, within a protein, are broken by a hydrolysis method. The degree of protein hydrolysis for the purposes of characterising the hydrolysed protein component of the composition is easily determined by one of ordinary skill in the formulation arts, by quantifying the amino nitrogen to total nitrogen ratio (AN/TN) of the protein component of the selected composition. The amino nitrogen component is quantified by USP titration methods for determining amino nitrogen content, with the total nitrogen component being determined by the Kjeldahl method. These methods are well-known to one of ordinary skill in the analytical chemistry art.

[0033] The term "partially hydrolysed", in terms of the present disclosure, means having a degree of hydrolysis which is greater than 0% but less than about 40%.

[0034] The term "extensively hydrolysed", in terms of the present disclosure, means having a degree of hydrolysis which is greater than or equal to about 40%.

[0035] The term "enteral' means deliverable through or within the gastrointestinal, or digestive, tract. "Enteral administration" includes oral feeding, intragastric feeding, transpyloric administration, or any other administration into the digestive tract. "Administration" is broader than "enteral administration" and includes parenteral administration or any other route of administration by which a substance is taken into a subject's body.

[0036] The term "functional milk", in terms of the present disclosure, means a functional milk-based beverage. A functional milk can impart a physiological benefit that enhances a subject's overall health, help prevent or treat a disease/condition in a subject, and/or improve a subject's physical or mental performance i.e. via an added functional ingredient, processing modification, and/or biotechnology. Examples of functional milks include liquid meal replacements, high-protein whey drinks, fortified milks, fermented milks, vitamin-enriched milks, and any other suitable milk-based beverage.

[0037] The term "functional water, in terms of the present disclosure, means a functional water-based beverage. A functional water can impart a physiological benefit that enhances a subject's overall health, help prevent or treat a disease/condition in a subject, and/or improve a subject's physical or mental performance i.e. via an added functional ingredient, processing modification, and/or biotechnology. Examples of functional waters include energy drinks, sports drinks, probiotic drinks, vitamin-infused water, electrolyte water, and any other suitable water-based beverage.

[0038] The term "substantially free" means containing less than a functional amount of the specified component, typically less than 0.1% by weight, and includes 0% by weight of the specified ingredient.

[0039] The term "adult', in terms of the present disclosure, means a human subject aged 18 years or older. The term "young adult" may refer to an adult aged between 18 and 35 years of age. The term "middle-aged adult' may refer to an adult aged between 36 and 55 years of age. The term "elderly adult' may refer to an adult aged over 55 years of age.

[0040] The term "exercise program", in terms of the present disclosure, refers to one or more physical activities undertaken to improve a subject's fitness and/or wellbeing.

[0041] The term "concurrent exercise program", in terms of the present disclosure, refers to the situation where a subject undertakes an exercise program whilst also undertaking an additional activity. This additional activity may be the regular (e.g. daily, weekly, etc.) consumption of the composition defined in the claims.

[0042] "Union", in terms of the present disclosure, refers to the combination of all outcomes related to physical performance i.e. the time to walk 400 metres (walking time), handgrip strength, and, the short physical performance battery (SPPB; i.e. standing balance (balance test), four metre walking speed (gait speed), and five repeated chair stands).

[0043] All percentages, parts, and ratios as used herein are detailed by weight of the total composition, unless otherwise specified. All amounts specified as administered "per day' may be delivered in a single unit dose, in a single serving, or in two or more doses or servings administered over the course of a 24-hour period.

[0044] All references to singular characteristics or limitations in the present disclosure shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary, by the context in which the reference is made.

[0045] The compositions of the present disclosure can comprise, consist of, or consist essentially of any of the components described herein, as well as including any additional useful component.

Detailed Description

[0046] The present invention provides compositions for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function. More particularly, the present invention provides compositions for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject.

[0047] Sarcopenia is a form of age-related loss of muscle mass and/or muscle function. The inventors of the present application have surprisingly found that different physiological pathways are involved in the development and/or regulation of sarcopenia in males, compared to those of females (see 'Experimental Procedure' section, Study 1). The inventors have determined that the most important physiological pathways in the development and/or regulation of sarcopenia in males and females are shown in Tables 1 and 2, respectively.

Table 1-differentially regulated pathways based on gene expression profiles of muscle biopsies in males Pathway Involved In Associated With Protein translation (eIF2-and el F4-dependent translational control) mTOR signalling, regulation of elF4 and p70S6K signalling, el F2 signalling Walking time, union Oxidative phosphorylation Oxidative phosphorylation, Grip strength, union mitochondrial dysfunction Cellular regulation of iron deficiency Iron homeostasis signalling pathway Grip strength, union Abbreviations: elF2 = eukaryotic initiation factor 2; elF4 = eukaryotic initiation factor 4; mTOR = mammalian/mechanistic target of rapamycin; p70S6K = ribosomal protein S6 kinase beta-1 Table 2 -differentially regulated pathways based on gene expression profiles of muscle biopsies in females Pathway Involved In Associated With NE-KB-dependent inflammation iNOS signalling, viral NF-kB activation, PKA signalling, T2D signalling, arginine biosynthesis Gait speed, union, VDR/RXR-dependent growth VDR/RXR activation SPPB, chair regulation stands, union Phosphatidylcholine biosynthesis Phosphatidylcholine metabolism Chair stands, balance test, union Folate receptor (FOLR)-related Folate binding and/or internalisation Gait speed, SPPB, chair stands, walking speed, union Abbreviations: NF-KB = nuclear factor kappa B; VDRJRXR = vitamin D receptor/retinoid X receptor; iNOS = inducible nitric oxide synthase; PKA = protein kinase cyclic adenosine monophosphatedependent; T2D = type 2 diabetes; SPPB = short physical performance battery.

[0048] As can be seen from Tables 1 and 2, the physiological pathways associated with the development and/or regulation of sarcopenia in males are significantly different to those of females, with no degree of overlap between males and females. Thus, it is clear that, in order to effectively treat, reduce, and/or prevent sarcopenia, a gender-specific intervention approach should be taken.

[0049] As discussed previously, the current recommendation for adults with sarcopenia is a combination of protein supplementation and physical exercise. When considering the physiological pathways responsible for the development and/or regulation of sarcopenia in males, protein supplementation may especially be a useful form of nutritional intervention.

However, in females, there is evidence that in addition to protein supplementation, other ingredients are required/critical based on the pathways responsible for the development and/or regulation of sarcopenia.

[0050] Thus, the inventors of the present application hypothesise that, in order to effectively treat, reduce, and/or prevent age-related loss of muscle mass and/or muscle function in females, nutritional interventions that have been shown, or are believed, to affect/interact with one or more of the four physiological pathways detailed in Table 2 are required. Further, it is hypothesised that a composition comprising a combination of nutritional ingredients, each of which target a different and/or the same physiological pathway, will result in a synergistic effect in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject.

[0051] To determine suitable nutritional intervention ingredients, the inventors conducted a literature search to identify ingredients that have been shown, or are believed, to affect/interact with each of the physiological pathways detailed in Table 2. The different ingredients identified as potential nutritional interventions, for each physiological pathway, are detailed below.

NF-KB-Dependent Inflammation [0052] Nuclear factor kappa B (NF-KB) is a key transcription factor that regulates a large array of genes involved in immune and inflammatory responses. NF-KB is located in the cell and can be activated by various pathogenic, stress, and danger signals. One function of the NF-KB signalling pathway is to organise a cellular (innate immune) response to these signals, and thus provide resistance to invading pathogens or damage. The NF-KB pathway has long been considered a prototypical pro-inflammatory signalling pathway, by regulating the expression of pro-inflammatory genes including cytokines, chemokines, and adhesion molecules. Deregulated NE-KB activation is a hallmark of chronic inflammatory diseases. In addition, pro-inflammatory cytokines are some of the most important inducers of muscle-wasting in chronic disease states, and have been suggested to play a role in age-related muscle loss and function (Li et al., J. Mol. Med., 2008).

[0053] The inventors identified NF-KB-dependent inflammation as a female-specific pathway in the development and/or regulation of age-related loss of muscle mass and/or muscle function, which is significantly associated with improved gait speed and union (i.e. the combination of physical performance exercises) outcomes.

Polyunsaturated Fatty Acids [0054] Docosahexaenoic Acid (DHA) and/or Eicosapentaenoic Acid (EPA) [0055] DHA and EPA are important polyunsaturated omega-3 fatty acids for humans as they are known to have beneficial effects on a number of physiological processes, such as foetal development, cardiovascular function, and Alzheimer's disease. DHA and/or EPA are typically obtained via the diet, with an important dietary source of DHA and/or EPA being fish oil.

[0056] DHA and EPA have been found to attenuate levels of NE-KB in an Mdx mice (model of dystrophy) through FFA1 and FFA4 (Maciel Junior et a/., Anat., 2021). In addition, in C2C12 myotubes, Wang et a/. (Biomed Res. Int., 2013) found that DHA inhibits protein degradation by regulating the PPARy/NF-KB pathway.

[0057] Also, a clinical study found that omega-3 fatty acid supplementation (4 g/day), for eight weeks, decreased oxidative parameters (malondialdehyde (MDA), protein carbonyl content (PCc), 8-hydroxy-2'-deoxyguanosine (8-OHdG)), and inflammation (tumour necrosis factor alpha (TNF-a)), induced by acute strenuous physical exercise (Buonocore et al., 2020), in the participants. Additionally, Ramos-Campo at al. (Nutrients, 2020) found that 10 weeks of DHA and EPA supplementation, in endurance athletes, significantly lowered muscle soreness and markers of muscle damage (creatine phosphokinase (CPK) and lactate dehydrogenase 5 (LDH-5)), and inflammation (interleukin 1 beta (IL-1 p), interleukin 6 (IL-6)), after one err:Anticinduced muscle damage exercise training session, when compared to the placebo group.

[0058] Further, a study by Marques et al. (AppI. Phys. Nutr. Metab., 2015) showed that daily DHA-rich fish oil supplementation reduced markers of muscle damage and inflammatory disturbances induced by acute exercise. These markers included plasma inflammatory mediators (including IL-1)3, IL-6, and TNF-a), markers of muscle damage (creatine kinase and LDH-5), and neutrophil function (inter alia cytokine production, DNA fragmentation, and production of reactive oxygen species).

[0059] Based on the above-described scientific research, the inventors hypothesise that DHA and/or EPA will have an effect on NF-KB-dependent inflammation, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, DHA and/or EPA have never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

[0060] Arachidonic Acid (ARA) [0061] ARA is a polyunsaturated omega-6 fatty acid that ads as a precursor for several prostaglandins with potential roles in muscle anabolism.

[0062] Chen et al. (Food Funct., 2016) conducted an in vitro study that explored the effects of ARA on palmitic acid-induced inflammatory responses and insulin resistance, in C2C12 myotubes. ARA was shown to inhibit inflammation and impaired insulin action, which in tum was cancelled in myotubes transfected with an active IKB kinase-I3 plasmid.

[0063] Further, a study by Mitchell et al. (J. Appl. Phys., 2018) showed that ARA supplementation did not affect muscle protein synthesis or anabolic signalling, but showed greater ribosome biogenesis and a trend toward greater change in satellite cell content, 48 hours after exercise. This suggests ARA supplementation may augment muscle adaptation in the following days of recovery after exercise.

[0064] Based on the above-described scientific research, the inventors hypothesise that ARA will have an effect on NF-KB-dependent inflammation, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, ARA has never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

Polyphenols [0065] Resveratrol [0066] Resveratrol is a polyphenol known to have various physiological effects, such as anti-inflammatory, anti-cancer, and anti-ageing effects.

[0067] Olesen e al. (Exp. Gerontol., 2013) conducted a study in mice that showed resveratrol supplementation, for three months, prevented an age-associated increase in skeletal muscle protein carbonylation, indicating that resveratrol reduces age-associated inflammation.

[0068] Also, in an in vitro model in C2C12 myotubes, resveratrol ameliorated palmitateinduced inflammation by decreasing the generation of reactive oxygen species and attenuating the JNK/NF-kB pathway in a sirtuin-1-independent mechanism, which suggests resveratrol could be a promising therapy for the prevention of inflammation in skeletal muscle cells (Sadeghi et al., J. Cell. Biochem., 2017).

[0069] Additionally, a rat study by Dolinsky et al. (J. Physiol., 2012) showed that resveratrol supplementation during exercise preserves protein balance, soleus muscle mass, and muscle maximal force contraction, as well as preserving mitochondria! capacity in unloaded muscle.

[0070] Further, Montesano et al. (J. Trans). Med., 2013) performed an in vitro study in C2C12 myotubes and found that resveratrol promoted myogenesis and hypertrophic processes, specifically by stimulating the insulin-like growth factor 1 (IGF-1) signalling pathway and AMP-activated protein kinase (AMPK) levels, as well as inducing hypertrophic morphological changes via the modulation of cytoskeletal protein expression.

[0071] In addition, a clinical study (Alway et al., J. Gerontol., 2017) showed that resveratrol, in combination with exercise, improved indices of mitochondria! density, muscle fatigue resistance, mean muscle fibre area, and total myonuclei in the elderly participants, to a greater extent than the placebo group or exercise alone.

[0072] Furthermore, a recent pilot clinical study by Harper et al. (Exp. Gerontol., 2021) demonstrated that resveratrol, combined with exercise, is safe for elderly adults with functional limitations and may improve mitochondria! function and mobility-related indices of physical function.

[0073] Based on the above-described scientific research, the inventors hypothesise that resveratrol will have an effect on NF-KB-dependent inflammation, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, resveratrol has never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

[0074] Flavonoids [0075] Flavonoids are a class of polyphenolic secondary metabolites found in plants. Nutritional sources that are rich in flavonoids are grape seeds and green tea. Extracts of green tea and grape seeds have shown powerful antioxidant and anti-inflammatory properties. Quercetin is a flavonoid present in garlic, grapes, tea, and other food sources, and has previously been shown to have antioxidant, anti-inflammatory and anti-cancer properties.

[0076] In a mice study by Lynn et al. (Nutrients, 2017), both quercetin and green tea extract supplementation induced a downregulation of immune and inflammatory response in soleus muscle tissue.

[0077] Evans et at (Clin Nutr, 2010) showed that green tea extract supplementation specifically reduced NF-KB staining in a Duchenne muscular dystrophy mouse model, which suggests that green tea extract can target the NE-KB-dependent inflammation in skeletal muscle.

[0078] In a mice study by Le et at (Mediators Inflamm., 2014), quercetin supplementation diminished the transcript levels of inflammatory receptors and the activation of their signalling molecules (extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinase (p38 MAPK), and NE-KB) in cocultured myotubes of high fat diet-fed obese mice. In addition, quercetin supplementation reduced expression of atrophic factors in the mice. This suggests that quercetin can target the NE-KB-dependent inflammatory and atrophic pathways in skeletal muscle.

[0079] Davis et at (Curr. Sports Med. Rep., 2009) showed that a seven-day dietary quercetin supplementation increases VO2 max and endurance capacity, without exercise intervention, in untrained participants. Other studies (Bazzucchi et at, Nutrients, 2020; Patrizio et al., Eur. J. Appl. Phys., 2020) found that quercetin supplementation improves neuromuscular performance, during and after resistance training, suggesting that quercetin supplementation may be beneficial for age-related loss of muscle mass.

[0080] Based on the above-described scientific research, the inventors hypothesise that flavonoids, such as quercetin, and food extracts rich in flavonoids, such as grape seed extract and green tea extract, will have an effect on NF-KB-dependent inflammation, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, flavonoids, such as quercetin, and flavonoid-rich extracts, such as green tea extract and grape seed extract, have never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

[0081] Curcumin [0082] Curcumin is a polyphenol compound and is most commonly known as the active component of turmeric (curcuma longa).

[0083] Singh & Aggarwal (J. Bio. Chem., 1995) showed that curcumin suppressed the activation of NF-kB in myeloid ML-la cells treated with TNF. Additionally, He at al. (Endoc., Metab. & Immun. Disord. Drug 2016) found that NF-kB/p65 protein abundance and calpain-, cathepsin L, and proteasome-dependent muscle proteolysis were decreased in mice after curcumin supplementation. This suggests curcumin is able to alter the inflammatory response during muscle proteolysis, such as in sarcopenia. Also, Manas-Garcia et al. (Nutrients, 2020) exhibited how curcumin supplementation, during muscle reloading in mice, decreased NF-kB p50 and proteolytic signalling markers. Further, Ono et al. (Exp. Phys., 2015) showed that curcumin injected into C57BL/6 J mice decreased skeletal muscle atrophy by inhibiting protein ubiquitination, inflammatory cytokines, and oxidative stress. This indicates that curcumin may be beneficial for treating muscle atrophy in type 1 diabetes patients. In summary, these studies determined that curcumin supplementation is able to decrease the inflammatory response, and proteolytic processes, involved in muscle atrophy via the NE-KB pathway.

[0084] Furthermore, a clinical study by Varma at al. (J. Med. Food, 2021) showed that provision of a curcumin supplement significantly improved hand grip strength, anti-fatigue effects, and muscle protein management in healthy elderly participants.

[0085] Based on the above-described scientific research, the inventors hypothesise that curcumin will have an effect on NF-KB-dependent inflammation, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, curcumin has never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

VDR/RXR-Dependent Growth Regulation [0086] The vitamin D receptor (VDR) is a nuclear, ligand-dependent transcription factor that functions as an obligate heterodimer with the retinoid X receptor (RXR; i.e. VDR/RXR), to regulate transcription. Binding of the active form of vitamin D3 to the VDR/RXR heterodimer, along with other co-activator proteins, mediates the transcriptional regulation of a number of vitamin D target genes (Christakos et a/., Physiol. Rev., 2015).

[0087] The inventors identified VDR/RXR-dependent activation as a female-specific pathway in the development and/or regulation of sarcopenia, which is significantly associated with short physical performance battery (SPPB), union, and chair stand outcomes.

[0088] Vitamin D [0089] Vitamin D3, the natural form of vitamin D, is an essential micronutrient. The synthesis of vitamin D3 by the skin upon ultraviolet (sunlight) exposure is the most important source of vitamin D. Vitamin D3 itself is not biologically active, but is metabolised by the liver to its active 25-hydroxyvitamin D3 (25(OH)D3) form, Mich also is a reliable biomarker of vitamin D status.

By binding to the vitamin D receptor, 25(OH)D3 activates a cascade of signals and mediators important for metabolic functions, neuromuscular transmission, and bone mineralisation (Kweder & Eidi, Avicenna J. Med., 2018).

[0090] An insufficient amount of vitamin D is a worldwide major public health problem, particularly amongst the elderly. A direct association between low 25(OH)D3 levels and frailty syndrome has been reported (Lunenfeld et al., Aging Male, 2015). In addition, a review by Latham et a/. (Front. Physiol., 2021) found that vitamin D may promote skeletal muscle regeneration and mitochondria! health.

[0091] Further, Abiri et al. (Methods Mol. Biol., 2020) showed that 12 weeks of vitamin D supplementation improves handgrip strength, timed 'get up and go', and body fat content in middle-aged women with a vitamin D deficiency.

[0092] Based on the above-described scientific research, the inventors hypothesise that vitamin D will have an effect on VDR/RXR-dependent growth regulation pathways, as well as a generally beneficial effect on health, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, vitamin D has never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

Phosphatidylcholine Biosynthesis From Choline [0093] Phosphatidylcholines are a class of phospholipids composed of a glycerol backbone esterified to phosphocholine and two fatty acids. Phosphatidylcholine inter alia is an essential structural component of cell membranes, in which it accounts for 40 to 60% of total phospholipids. While phosphatidylcholine has long been regarded primarily as a structural lipid and cellular building block, recent evidence has shown that ifs able to influence gene expression and homeostatic control of glucose regulation, via insulin transduction (Furse and de Kroon, Mol. Membr. Bio, 2015). In all nucleated mammalian cells, phosphatidylcholine is synthesised from choline via the Kennedy pathway, the main phosphatidylcholine biosynthesis pathway. Phosphatidylcholine can also be synthesised via the Lands cycle, or de novo biosynthesis from phosphatidylethanolamine via the phosphatidylethanolamine methyl transferase (PEMT) pathway.

[0094] Choline is primarily obtained from the diet and its main fate is the synthesis of phosphatidylcholine. Phosphatidylcholine biosynthesis from choline is important for maintenance of the phospholipid composition of mitochondria! membranes in muscle, and defects in this pathway can cause muscle pathology (van der Veen et aL, Biochim. Biophys., 2017). An abnormally high and low cellular phosphatidylcholine:phosphatidylethanolamine ratio influences energy metabolism and is linked to disease progression.

[0095] The inventors identified phosphatidylcholine biosynthesis from choline (Kennedy pathway) was a female-specific pathway in the development and/or regulation of sarcopenia, which is significantly associated with improved chair stand, balance test, and union outcomes.

Phosphatidylcholine Source [0096] Milk Fat Globule Membrane (MFGM) [0097] MFGM is a naturally occurring bioactive membrane structure that surrounds the fat droplets in human breast milk and other mammalian milk e.g. cow's milk. MFGM is comprised of a trilayer lipid structure that comprises a complex mixture of phospholipids, proteins, glycoproteins, triglycerides, polar lipids, cholesterol, enzymes, and other components. One of the phospholipids present in MFGM is phosphatidylcholine. MFGM is therefore a useful source of phosphatidylcholine.

[0098] In newborns, phosphatidylcholine in MFGM is an important source of choline. MFGM supplementation in infants has shown to increase circulating concentrations of phosphatidylcholine, as well as erythrocyte membrane levels of phosphatidylcholine (Grip et al., Pediatr. Res., 2018). Krill oil is also high in phosphatidylcholine and, as discussed in further detail below, krill oil supplementation results in increased plasma choline levels (Bjorndal et al., Scand. J. Clin., 2018). The inventors hypothesise that the provision of a composition comprising MFGM will lead to an increased muscle supply of phosphatidylcholine, and/or choline for phosphatidylcholine biosynthesis, which will lead to the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject.

[0099] Genetic defects in phosphatidylcholine biosynthesis are linked to impaired muscle function. Pathogenic variants in the CHKB gene, which encodes for an enzyme in phosphatidylcholine synthesis, causes the mice to develop the mice equivalent of congenital muscular dystrophy (Sher et al., J. Biol. Chem., 2006; Misuhashi et al., Am. J. Hum. Genet., 2011).

[0100] In elderly individuals, phosphatidylcholine levels in muscle were elevated in sarcopenic muscle and were inversely linked to muscle volume and peak power (Hinkley et al., Aging Cell, 2020). Also, plasma choline levels were inversely associated with sarcopenia (Lu et al., J. Gerontol., 2020).

[0101] As summarised by Raza et al. (J. Dairy Sci., 2021), there are at least six studies that show MFGM supplementation (typically 1 g/day) improved mobility in older/elderly adults (Kim et al., PLoS One, 2015; Ota et al., SpringerPlus, 2015; Minegishi et al., J. Nutr. Sci. Vitaminol., 2016; Kokai et al., J. Nutr. Sci., 2018; Yoshinaka et at, Biosci. Biotechnol. Biochem., 2018; Kim et aL, Arch. Gerontol. Geriatr., 2019) [0102] Based on the above-described scientific research, the inventors hypothesise that MFGM will have an effect on the phosphatidylcholine biosynthesis pathway, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, MFGM has never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

[0103] Krill Oil [0104] Krill oil is an oil extracted from shrimp-like crustaceans called krill, and is often used as a supplement as it is rich in phosphatidylcholine and astaxanthin, as well as the polyunsaturated omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA).

[0105] In a human pilot study by Bjorndal et a/. (Scand. J. Clin., 2018), krill oil supplementation increased fasting plasma choline levels and other metabolites along the choline oxidation pathways such as betaine, dimethylglycine, and sarcosine. In another human pilot study, Georges et al. (J. Nutr. Metab., 2018) found that krill oil supplementation, in combination with an eight-week resistance training program, significantly stimulated mTOR signalling and increased lean body mass in the participants.

[0106] Further, a study by Hinkley et at (Aging Cell, 2020) in obese adults showed that phosphatidylcholine levels were elevated in sarcopenic muscle and were inversely related to muscle volume and peak power. This suggests an association between phospholipids, such as phosphatidylcholine, and loss of skeletal muscle mass and function in adults.

[0107] Based on the above-described scientific research, the inventors hypothesise that krill oil will have an effect on phosphatidylcholine biosynthesis-related pathways, and therefore be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, krill oil has never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

Folate Receptor-Related [0108] Folate receptors are a class of membrane-bound surface proteins that bind folate, folic acid, and other folate forms/derivatives with high affinity. The folate receptor functions to capture and internalise extracellular folates via endocytosis. After uptake by the folate receptor, folate is converted to tetrahydrofolate, which serves as a backbone for one-carbon reactions. Thereby, folate is a basic component of cell metabolism and DNA synthesis and repair. In vitro studies have shown that expression of the folate receptor increases when extracellular levels of folate are low, possibly through changes in intracellular folate concentrations. In this way, the folate receptor is important for cellular folate acquisition and cellular survival. The (regulation of the) folate receptor has widely been studied in tumour cells and cancer development, but its role in muscle (cells) and sarcopenia remains largely unclear.

[0109] The inventors identified the folate receptor as a regulator that specifically in females is significantly associated with improved gait speed, SPPB, chair stand, walking speed, and union.

[0110] Folate and/or Folic Acid [0111] Folate is the natural form of vitamin B9 in food. Folic acid is a synthetic form of folate and is usually added to supplements and processed foods. Folate is important to limit homocysteine concentrations, reduce inflammation, and contribute to mitochondria! functioning. The body requires about 400pg of folate/folic acid daily, but cannot create folate or folic acid, so folate and/or folic acid must be obtained via the diet.

[0112] Lee et at determined that folate levels are related to muscle strength in Korean population, especially in elderly (Nutrients, 2021). Additionally, in a clinical study involving a relatively folate-deficient study population, serum levels of folate were significantly associated with muscle strength, especially in women (Wee et at, Nutr. J., 2016). Further, Ao et a/. found that blood folate concentration was significantly associated with handgrip strength and physical performance (J. Nutr. Sci. Vitaminol., 2019). Also, interestingly, a positive association between serum folate levels and grip strength, in women but not men, was identified by Zhang et al. (NHANES data;, Arch. Osteoporosis, 2021). Furthermore, A lower intake of folate was associated with higher risk of frailty (Balbao-Castillo at al., Age Ageing, 2018).

[0113] Based on the above-described scientific research, the inventors hypothesise that the upregulation of folate receptor pathways in females reflects low intracellular folate levels. Therefore, folate and/or folic acid will be a useful ingredient in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in females. To the best of the inventors' knowledge, folate and/or folic acid have never previously been identified as a female-specific intervention for age-related loss of muscle mass and/or muscle function.

[0114] In one aspect, the present application provides a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises at least one of the following: DHA, EPA, resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, arachidonic acid (ARA), krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof.

[0115] In another aspect, there is provide a composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject comprising: * at least one of the following: DHA, EPA, resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, ARA, krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof; and, * at least one of the following: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof.

[0116] In another aspect, there is provided a composition comprising at least two of the following: * at least one of DHA, EPA, ARA, or any combination thereof; * resveratrol; * quercetin, green tea extract, and/or grape seed extract; * curcumin; * MFGM and/or krill oil; * vitamin D; and, * folate and/or folic acid.

[0117] In another aspect, there is provided a composition comprising: * at least two of the following: o at least one of DHA, EPA, ARA, or any combination thereof; o resveratrol; o quercetin, green tea extract, and/or grape seed extract; o curcumin; o MFGM and/or krill oil; o vitamin D; and, o folate and/or folic acid; and, * at least one of the following: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof [0118] In another aspect, there is provided a composition for use in the treatment, reduction, or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises at least two of the following: * at least one of DHA, EPA, ARA, or any combination thereof; * resveratrol; * quercetin, green tea extract, and/or grape seed extract; * curcumin; * MFGM and/or krill oil; * vitamin D; and, * folate and/or folic acid.

[0119] In another aspect, there is provided a composition for use in the treatment, reduction, or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises: * at least two of the following: o at least one of DHA, EPA, ARA, or any combination thereof; o resveratrol; o quercetin, green tea extract, and/or grape seed extract; o curcumin; o MFGM and/or krill oil; o vitamin D; and, o folate and/or folic acid; and, * at least one of the following: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof [0120] In another aspect, there is provided a method for the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, the method comprising providing a composition comprising at least one of the following: DHA, EPA, resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, ARA, krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof.

[0121] In another aspect, there is provided a method for the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, the method comprising providing a composition comprising at least one of the following: DHA, EPA, resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, ARA, krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof, in the absence of a concurrent exercise program.

[0122] In another aspect, there is provided a method for the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, the method comprising providing a composition comprising at least one of the following: DHA, EPA, resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, ARA, krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof, in the presence of a concurrent exercise program.

[0123] In any aspect detailed above, the age-related loss of muscle mass and/or muscle function may comprise sarcopenia. Alternatively, in any aspect detailed above, the age-related loss of muscle mass and/or muscle function is sarcopenia.

[0124] The composition of any aspect detailed above may comprise docosahexaenoic acid (DHA) and/or eicosapentaenoic acid (EPA). The DHA and/or EPA may be present in the composition in an amount of 100 milligrams (mg) to 2500 mg. Preferably, the DHA and/or EPA is present in the composition in an amount of 250 mg to 2000 mg. More preferably, the DHA and/or EPA is present in the composition in an amount of 400 mg to 1200 mg. Most preferably, the DHA and/or EPA is present in the composition in an amount of 500 mg to 1000 mg. The daily dosage of DHA and/or EPA may be 250 mg to 2000 mg. Preferably, the daily dosage of DHA and/or EPA is 500 mg to 1000 mg. The amounts and daily dosages of DHA and/or EPA detailed may refer to each ingredient individually, or to the ingredients collectively.

[0125] The composition of any aspect detailed above may comprise resveratrol. The resveratrol may be present in the composition in an amount of 5 mg to 750 mg. Preferably, the resveratrol is present in the composition in an amount of 10 mg to 500 mg. More preferably, the resveratrol is present in the composition in an amount of 25 mg to 250 mg. Most preferably, the resveratrol is present in the composition in an amount of 50 mg to 150 mg. The daily dosage of resveratrol may be 100 mg to 2000 mg. Preferably, the daily dosage of resveratrol is 150 mg to 250 mg.

[0126] The composition of any aspect detailed above may comprise MFGM. The MFGM may be present in the composition in an amount of 25 mg to 1000 mg. Preferably, the MFGM is present in the composition in an amount of 50 mg to 500 mg. More preferably, the MFGM is present in the composition in an amount of 75 mg to 250 mg. Most preferably, the MFGM is present in the composition in an amount of 100 mg to 200 mg. The daily dosage of MFGM may be 100 mg to 2000 mg. Preferably, the daily dosage of MFGM is 200 mg to 1000 mg.

[0127] The composition of any aspect detailed above may comprise vitamin D. The vitamin D may be present in the composition in an amount of 5 micrograms (pg) to 200 pg. Preferably, vitamin D is present in the composition in an amount of 10 pg to 100 pg. More preferably, the vitamin D is present in the composition in an amount of 15 pg to 75 pg. Most preferably, the vitamin D is present in the composition in an amount of 20 pg to 50 pg. The daily dosage of vitamin D may be 10 pg to 100 pg. Preferably, the daily dosage of vitamin D is 20 pg to 50 pg.

[0128] The composition of any aspect detailed above may comprise folate and/or folic acid. The folate and/or folic acid may be present in the composition in an amount of 50 pg to 1500 pg. Preferably, the folate and/or folic acid is present in the composition in an amount of 100 pg to 1000 pg. More preferably, the folate and/or folic acid is present in the composition in an amount of 200 pg to 800 pg. Most preferably, the folate and/or folic acid is present in the composition in an amount of 300 pg to 500 pg. The daily dosage of folate and/or folic acid may be 100 pg to 1000 pg. Preferably, the daily dosage of folate and/or folic acid is 300 pg to 500 pg. The amounts and daily dosages of folate and/or folic acid detailed may refer to each ingredient individually, or to the ingredients collectively.

[0129] The composition of any aspect detailed above may comprise ARA. The ARA may be present in the composition in an amount of 200 mg to 1000 mg. Preferably, the ARA is present in the composition in an amount of 250 mg to 800 mg. More preferably, the ARA is present in the composition in an amount of 350 mg to 775 mg. Most preferably, the ARA is present in the composition in an amount of 500 mg to 750 mg. The daily dosage of ARA may be 500 mg to 1600 mg. Preferably, the daily dosage of ARA is 1000 mg to 1500 mg.

[0130] The composition of any aspect detailed above may comprise krill oil. The krill oil may be present in the composition in an amount of 100 mg to 1500 mg. Preferably, the krill oil is present in the composition in an amount of 200 mg to 1000 mg. More preferably, the krill oil is present in the composition in an amount of 225 mg to 750 mg. Most preferably, the krill oil is present in the composition in an amount of 250 mg to 500 mg. The daily dosage of krill oil may be 200 mg to 4000 mg. Preferably, the daily dosage of krill oil is 500 mg to 1000 mg.

[0131] The composition of any aspect detailed above may comprise one or more flavonoids and/or one or more flavonoid-containing food extracts. The one or more flavonoids may comprise quercetin. The one or more flavonoid-containing food extracts may comprise green tea extract, grape seed extract, or a combination thereof. The one or more flavonoids and/or one or more flavonoid-containing food extracts may be present in the composition in an amount of 25 mg to 1500 mg. Preferably, the one or more flavonoids and/or one or more flavonoid-containing food extracts are present in the composition in an amount of 50 mg to 1000 mg. More preferably, the one or more flavonoids and/or one or more flavonoid-containing food extracts are present in the composition in an amount of 75 mg to 750 mg. Most preferably, the one or more flavonoids and/or one or more flavonoid-containing food extracts are present in the composition in an amount of 100 mg to 500 mg. The daily dosage of one or more flavonoids and/or one or more flavonoid-containing food extracts may be 50 mg to 1000 mg. Preferably, the daily dosage of one or more flavonoids and/or one or more flavonoid-containing food extracts is 100 mg to 500 mg. The amounts and daily dosages of one or more flavonoids and/or one or more flavonoid-containing food extracts detailed may refer to each ingredient individually, to a combination of two of the ingredients, or to the ingredients collectively.

[0132] The composition of any aspect detailed above may comprise curcumin. The curcumin may be present in the composition in an amount of 10 mg to 3000 mg. Preferably, the curcumin is present in the composition in an amount of 40 mg to 2500 mg. More preferably, the curcumin is present in the composition in an amount of 100 mg to 2000 mg. Most preferably, the curcumin is present in the composition in an amount of 400 mg to 1000 mg. The daily dosage of curcumin may be 150 mg to 2500 mg. Preferably, the daily dosage of curcumin is 400 mg to 1000 mg.

[0133] The composition of any aspect detailed above may comprise a protein source. The protein source provides protein to the composition. The protein source may comprise intact protein, partially hydrolysed protein, extensively hydrolysed protein, small amino acid peptides, or any combination thereof. The protein source may be derived from any mammalian milk protein or plant protein, as well as their fractions, or any combination thereof. The protein source may comprise bovine milk, caprine milk, whey protein, casein protein, soy protein, rice protein, pea protein, peanut protein, egg protein, sesame protein, fish protein, wheat protein, hydrolysed protein, or any combination thereof. Bovine milk protein sources may comprise, but are not limited to, milk protein powders, milk protein concentrates, milk protein isolates, non-fat milk solids, non-fat milk, non-fat dry milk, whey protein, whey protein isolates, whey protein concentrates, sweet whey, acid whey, casein, acid casein, caseinate (e.g. sodium caseinate, sodium calcium caseinate, calcium caseinate), or any combination thereof. Preferably, the protein source comprises whey protein.

[0134] When the composition of any aspect detailed above is in the form of a powder, the protein source may be present in the composition in an amount of about 5 g/100 g of powder to about 99 g/100 g of powder. Preferably, the protein source is present in an amount of about 10 g/100 g of powder to about 75 g/100 g of powder. More preferably, the protein source is present in an amount of about 15 g/100 g of powder to about 50 g/100 g of powder.

[0135] When the composition of any aspect detailed above is in the form of a liquid, the protein source may be present in the composition in an amount of about 0.1 g/100 mL of powder to about 20 g/100 mL. Preferably, the protein source is present in an amount of about 0.5 g/100 mL to about 15 g/100 mL. More preferably, the protein source is present in an amount of about 1 g/100 mL to about 10 g/100 mL.

[0136] The composition of any aspect detailed above may comprise a milk source. The milk source provides milk to the composition. The milk source may be a mammalian milk source.

The mammalian milk source may be derived from bovine milk, caprine milk, ovine milk, or any combination thereof. The milk source may comprise whole milk, whole milk powder, lactose-free whole milk, lactose-free whole milk powder, whole milk concentrates, whole milk isolates, lactose-free whole milk isolates, whole milk solids, semi-skimmed (reduced fat) milk, semi-skimmed (reduced fat) milk powder, lactose-free semi-skimmed (reduced fat) milk, lactose-free semi-skimmed (reduced fat) milk powder, semi-skimmed (reduced fat) milk concentrates, semi-skimmed (reduced fat) milk isolates, lactose-free semi-skimmed (reduced fat) milk isolates, semi-skimmed (reduced fat) milk solids, skimmed (non-fat) milk, skimmed (non-fat) milk powder, lactose-free skimmed (non-fat) milk, lactose-free skimmed (non-fat) milk powder, skimmed (non-fat) milk concentrates, skimmed (non-fat) milk isolates, lactose-free skimmed (non-fat) milk isolates, skimmed (non-fat) milk solids, buttermilk, or any combination thereof.

[0137] The milk source may be an altemative milk source. The alternative milk source may be any non-dairy, vegan-friendly, and/or plant-based altemative milk source. The alternative milk source may comprise coconut milk, soy milk, almond milk, oat milk, hazelnut milk, pea milk, potato milk, cashew milk, quinoa milk, an animal-free milk, or any combination thereof. The alternative milk source may comprise a whey protein produced by microflora, a whey protein produced from amino acids, a casein protein produced by microflora, a casein protein produced from amino acids, or any combination thereof.

[0138] The milk source may be subjected to fermentation with one or more species of bacteria, to produce a fermented milk source. The milk source may therefore be a fermented mammalian milk source, a fermented alternative milk source, or a combination thereof.

[0139] The composition of any aspect detailed above may comprise a source of one or more free amino acids. The source of one or more free amino acids provides free amino acids to the composition. The source of one or more free amino acids may comprise alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, or any combination thereof. When the composition is in the form of a powder, the source of one or more free amino acids may be present in the composition in an amount of about 5 g/100 g of powder to about 99 g/100 g of powder. Preferably, the source of one or more free amino acids is present in an amount of about 25 g/100 g of powder to about 99 g/100 g of powder. More preferably, the source of one or more free amino acids is present in an amount of about 50 g/100 g of powder to about 99 g/100 g of powder. The amounts of the source of one or more free amino acids detailed may refer to a single free amino acid, or to a combination of two or more free amino acids.

[0140] It is hypothesised that the composition of any aspect detailed above will exert a beneficial effect in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, in the absence of a concurrent exercise program. The composition of any aspect detailed above may be effective in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, when provided in the absence of an exercise program.

[0141] It is hypothesised that the composition of any aspect detailed above will exert a synergistic beneficial effect in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, when combined with a concurrent exercise program. The composition of any aspect detailed above may be effective in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, when provided in the presence of a concurrent exercise program.

[0142] The composition of any aspect detailed above is intended for a female adult. Preferably, the female adult is a young adult, a middle-aged adult and/or an elderly adult. More preferably, the female adult is a middle-aged adult and/or an elderly adult.

[0143] The composition of any aspect detailed above may be a supplement, a nutritional composition, or a combination thereof. The nutritional composition may be a nutritional supplement, a functional milk, a functional water, or any other nutritional composition.

[0144] The composition of any aspect detailed above may be provided in the form of a powder, a gel, a suspension, a paste, a solid, a liquid, a liquid concentrate, a reconstitutable powder, a reconstituted solution, or a ready-to-use product. Preferably, the composition is in the form of a reconstitutable powder, a reconstituted solution, or a ready-to-use product. Most preferably, the composition is provided in the form of a reconstitutable powder.

[0145] The composition of any aspect detailed above may be provided in an orally-ingestible form, wherein the orally-ingestible form comprises a food, a beverage, a tablet, a capsule, a gummy, or a powder. The composition may be expelled directly into a subjects intestinal tract. The composition may be expelled directly into a subject's gut. The composition may be formulated to be consumed or administered enterally under the supervision of a physician.

[0146] The daily dosage of the composition of any aspect detailed above may be provided in the form of a single dosage. The single dosage may comprise a single discrete unit of the composition (i.e. a single tablet, capsule, gummy, powder amount, liquid volume, gel serving, suspension volume, paste amount, liquid concentrate amount, reconstitutable powder amount, reconstituted solution volume, or ready-to-use product) or multiple discrete units of the composition (i.e. two or more tablets, capsules, gummies, gel servings, or ready-to-use products). Alternatively, the daily dosage of the composition of any aspect detailed above may be provided in the form of multiple doses e.g. at two points in the same day, at three points in the same day, etc. The multiple daily doses may each comprise a single discrete unit of the composition (i.e. a single tablet, capsule, gummy, powder amount, liquid volume, gel serving, suspension volume, paste amount, liquid concentrate amount, reconstitutable powder amount, reconstituted solution volume, or ready-to-use product) or multiple discrete units of the composition (i.e. two or more tablets, capsules, gummies, gel servings, or ready-to-use products).

[0147] In any aspect detailed above, the treatment and/or reduction of age-related loss of muscle mass and/or muscle function in a female subject may refer to an increase in muscle strength (i.e. grip strength, lower body strength) and/or muscle performance (i.e. gait speed, chair stand time test), in the presence of the composition of any aspect detailed above, of at least 1%, compared to a control value in the absence of the composition of any aspect detailed above; preferably, an increase of at least 5%; more preferably, an increase of at least 10%; even more preferably, an increase of at least 20%; most preferably, an increase of at least 50%. Muscle strength and/or muscle performance may be quantified by one or more of the methods detailed below, or any other suitable method known in the art: * Muscle strength o Grip strength Measured using a digital dynamometer in a sitting position, with the maximum grip strength expressed in kg. Three repetitions are performed with the right hand, then three repetitions with the left hand, with the mean of the two best for each hand recorded.

o Lower body strength (i) Muscle peak torque: measurement of peak isometric torque (Nm) and average peak isometric torque (Nm) (ii) Isometric knee muscle strength extension test * Muscle performance o Gait speed Participants are requested to walk through an 11 metre (m) straight course at a normal speed, with the time taken to walk between the 3 m and 8 m marks of the course recorded.

o Chair stand time test Subjects stand up from a chair, walk 3 m, turn around, return, and sit on the chair as quickly as possible, with the time taken recorded.

[0148] The scope of the present application is defined in the appended claims. It is to be understood that the skilled person may make amendments to the scope of the claims without departing from the scope of the present disclosure.

Experimental Procedure Study 1: Determination of the physiological pathways involved in the development of sarcopenia in males and females [0149] A cross-sectional study was performed wherein adults (75 years old and older) were screened for (pre-)frailty using the Fried criteria (weight loss, exhaustion, low physical activity, weakness, and slowness) and assigned to a 'fit' group (no indications of frailty; 15 males, 15 females) or a (pre-)frail' group 1 indicator of frailty; 13 males, 12 females). Healthy, young participants (20 to 30 years old, 13 males, 13 females) with a healthy BM! (20 to 25 kg/m2) were included as a reference group. In total, 81 participants were included in the study.

[0150] On their first visit to the research facility, body composition and physical performance of the study participants were assessed. The body composition assessment involved measuring body weight, height, and bone density. The physical performance assessment was performed only on the 'fit' and 'frail' elderly participants and included: the time to walk 400 metres (walking time); handgrip strength; and, the short physical performance battery (SPPB) i.e. standing balance (balance test), four metre walking speed (gait speed), and five repeated chair stands. On their second visit, six weeks later, a blood sample and a muscle biopsy (from the vastus lateralis) were taken from the participants. RNA sequencing was used to quantify RNA expression in the muscle biopsies.

[0151] Analysis of the data obtained from the study revealed large differences in muscle RNA gene expression between males and females, in all three groups. This difference in RNA expression likely indicates that sarcopenia is regulated differently in males and females.

[0152] To identify the physiological pathways associated with sarcopenia, a pathway analysis was performed for each gender separately. Data from the gait speed test, walk test, and the chair stand test were transformed so that a positive correlation indicated a positive relationship between the expression of a gene and functional outcomes of physical activity, and a negative correlation indicated a negative relationship. For example, increased handgrip strength was regarded as a positive effect, as it would likely indicate increased (muscle) performance, whereas increased walking time was regarded as negative effect, as it likely indicates decreased (muscle) performance. Furthermore, an additional outcome was created, called union', which comprised those unique genes significantly correlated to outcomes related to physical performance i.e. all outcomes except lean mass.

[0153] The significantly correlating genes were then used as an input for pathway analysis through Ingenuity Pathway Analysis (IPA) suite. Significant pathways were identified by means of Fishers exact test, wherein a p-value 0.01 was considered significant. A similar analysis was performed to identify significant upstream regulators (UPRs).

[0154] The significant physiological gender-specific pathways and UPRs were selected using three different approaches, of which the outcomes were pooled to ensure that not only those with the highest p-values were selected, but also pathways or UPRs that were associated with multiple functional outcomes. The approaches described below were applied for males and females separately. A physiological pathway or U PR was considered significant if the -log (p-value) was ? 2.

[0155] Approach 1: The top 10 pathways with the highest p-values for 'union' were plotted in bar charts. All significant pathways were selected and included in the final list.

[0156] Approach 2: All pathways with the highest p-values per functional outcome, also referred to as all first place' pathways, were selected and included in the final list.

[0157] Approach 3: The top 10 pathways with the highest p-values for all functional outcomes (excluding union) were plotted in separate bar charts. The number of times a pathway occurred in a top 10 of a functional outcome was counted (including union). For example, if the pathway X was displayed in the top 10 for Y and Z, the count was 2. All pathways with a count ? 2 were selected and included in the final list.

[0158] In total, 16 pathways were selected using two approaches and four pathways using three approaches. The greater the number of times a pathway had been identified by an approach, the more it was considered to be an important and relevant pathway. The particular pathway was therefore assigned a higher 'priority status' during the subsequent selection steps. A similar selection procedure was applied for all UPRs, except for approach 3, wherein only the significant UPRs with a total count of ? 2 were selected.

[0159] The gender-specific pathways were then prioritised and ranked to select a top three list for each gender. To do this, firstly, the genes underlying the selected pathways were visualised, the pathways were grouped together to form clusters based on the number of overlapping genes underlying the pathway and were given an alternate, more comprehensive name, if deemed relevant. In addition, our own knowledge on their physiological function also played a role in the formation of the clustering pathways. The genes underlying the selected UPRs were also visualised. The UPRs were regarded as additional support for the pathways and were excluded if no overlap of the underlying genes with those of a pathway was found.

[0160] After clustering the pathways together to, where appropriate, form a more comprehensive pathway, the various pathways were ranked according to the number of pathways clustered together, their physiological relevance in the development/onset of sarcopenia based on our own knowledge, and the p-values of the separate underlying pathways.

[0161] The top three pathways for each gender are detailed in Tables 1 and 2 (see 'Detailed Description' section), along with their respective functional outcomes in the aforementioned clinical study. For females only, pathways involving the folate receptor (FOLR) were also included as it was determined to be an important female-specific UPR in the development and/or regulation of sarcopenia.

Study 2: Outline of study protocol for determining the efficacy of the compositions of the present invention in the treatment and/or reduction of age-related loss of muscle mass and/or muscle function in a female subject [0162] The study subjects would ideally be females (preferably 60 years and older) with decreased muscle strength e.g. low gait speed, walking speed <1m/s, and/or low handgrip strength <20kg. Any females taking one or more supplements with known effects on muscle health would be excluded from the study. The subjects would be randomly assigned to an intervention or a placebo study group, which each ideally consist of a minimum 25 subjects, preferably 50 subjects per study group. The subjects would then receive an intervention based on the present invention, or a placebo, daily for a minimum of eight weeks, preferably for a minimum of 24 weeks. Half of the subjects from the intervention study group and half of the subjects from the placebo study group would be asked to undertake 30 minutes of strength-training exercise, three times per week, with the other half of each study group asked not to change their usual exercise routine. All subjects would be provided with a personal diary to record their adherence to the study intervention.

[0163] The treatment and/or reduction of age-related loss of muscle mass and/or muscle function (e.g. sarcopenia) in a female subject may be assessed and quantified as previously described e.g. comparison of baseline (i.e. prior to study intervention) and immediate post-study intervention values for increase in muscle strength tests (e.g. grip strength, lower body strength) and/or muscle performance tests (e.g. balance, gait speed, chair stand time test), or by any other suitable method known in the art. Treatment effects will be tested using generalised linear models, which will be adjusted for baseline outcome values and confounding factors.

Example Compositions

[0164] The compositions shown in Table 3 illustrate examples of compositions within the scope of the present disclosure, but are in no way intended to provide any limitation on the disclosure.

Table 3

Component Composition

I II III IV V VI VII VIII IX X

DHA and/or EPA 1XX X X X X X X X Resveratrol X V X X X X X X X X Milk fat globule membrane (MFGM) X X V X X X X X X X Vitamin D XXX IX X X X X / Folate and/or folic acid XXX X / X X X X / ARA XXX X X / X X X X Krill oil XXX X X X / X X X Flavonoid (e.g. quercetin) XXX X X X X IX X and/or flavonoid-containing food extract (e.g. green tea and/or grape seed) Curcumin XXX X X X X X / X Key: / = present; X = not present Component Composition

XI XII XIII XIV XV XVI XVII XVIII XIX

DHA and/or EPA / / X / / X X / / Resveratrol X / / / X / X X / Milk fat globule membrane (MFGM) / X / / X X / / / Vitamin D X X X X / / / / / Folate and/or folic acid X X X X77 / 77

ARA XXXXX X X X X

Krill oil X X X X X X X X X Flavonoid (e.g. quercetin) X X X X X X X X X and/or flavonoid-containing food extract (e.g. green tea and/or grape seed) Curcumin XXXXX X X X X Key: / = present; X = not present

Claims

Claims 2. 3. 4. 6. 7.A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises at least one of the following: docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, arachidonic acid (ARA), krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof.
A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises DHA and/or EPA in an amount of 100 milligrams (mg) to 2500 mg.
A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises resveratrol in an amount of 5 mg to 750 mg.
A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises milk fat globule membrane (MFGM) in an amount of 25 mg to 1000 mg.
A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises vitamin D in an amount of 5 micrograms (pg) to 200 pg.
A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises folate or folic acid in an amount of 50 pg to 1500 pg.
A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises ARA in an amount of 200 mg to 1000 mg.
8. A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises krill oil in an amount of 100 mg to 1500 mg.
9. A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises quercetin, green tea extract, and/or grape seed extract in an amount of 25 mg to 1500 mg.
10. A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises curcumin in an amount of 10 mg to 3000 mg.
11. A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises vitamin D and at least one of folate, folic acid, or a combination thereof.
12. A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises DHA and/or EPA, vitamin D and at least one of folate, folic acid, or a combination thereof
13. A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject, wherein the composition comprises DHA and/or EPA, resveratrol, MFGM, vitamin D and at least one of folate, folic acid, or a combination thereof.
14. The composition for use according to any one of the preceding claims, wherein the age-related loss of muscle mass and/or muscle function comprises sarcopenia.
15. The composition for use according to any one of the preceding claims, wherein the composition is a supplement, a nutritional composition, or a combination thereof
16. The composition for use according to any one of the preceding claims, wherein the composition is a synthetic composition.
17. The composition for use according to any one of the preceding claims, wherein the female subject is a middle-aged adult or an elderly adult.
18. The composition for use according to any one of the preceding claims, wherein the daily dosage of the composition is provided in a single dosage.
19. A composition for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject comprising: * at least one of the following: DHA, EPA, resveratrol, milk fat globule membrane (MFGM), vitamin D, folate, folic acid, ARA, krill oil, quercetin, green tea extract, grape seed extract, curcumin, or any combination thereof; and, * at least one of the following: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof.
20. A composition comprising at least two of the following: * at least one of DHA, EPA, ARA, or any combination thereof; * resveratrol; * quercetin, green tea extract, and/or grape seed extract; * curcumin; * MFGM and/or krill oil; * vitamin D; and, * folate and/or folic acid.
21. A composition comprising: * at least two of the following: o at least one of DHA, EPA, ARA, or any combination thereof; o resveratrol; o quercetin, green tea extract, and/or grape seed extract; o curcumin; o MFGM and/or krill oil; o vitamin D; and, o folate and/or folic acid; and, * at least one of the following: a protein source, a source of one or more free amino acids, a milk source, or any combination thereof.
22. The composition of claim 20 or 21, for use in the treatment, reduction, and/or prevention of age-related loss of muscle mass and/or muscle function in a female subject.
23. The composition for use according to claim 22, wherein the age-related loss of muscle mass and/or muscle function comprises sarcopenia.