A method to treat body composition resistance

Abstract

A method of treating body composition resistance in a mammal is provided comprising administering to the mammal for a sufficient period of time nutrients comprising protein in an amount of 10-100g, creatine in an amount of 1-5g, vitamin D in an amount of 50-4000 IU, calcium in an amount of 100-1000 mg, and omega-3 fatty acids in an amount of 500-15,000 mg. In embodiments, the method additionally improves one or more of lean mass, muscle strength, body composition index, fat mass and % body fat, systemic inflammation levels, blood cholesterol levels and / or blood triglyceride levels

Description

A METHOD TO TREAT BODY COMPOSITION RESISTANCEField of the Invention

[0001] The present invention generally relates to obesity-associated body composition resistance (BCR), and in particular, to the treatment of body composition resistance in a mammal (human and non-human) with a multi-nutrient composition.Background of the Invention

[0002] A major threat to the health and economies of many countries is the growing obesity epidemic. Obesity is linked to debilitating comorbidities and premature death, with a 30 % increase in all-cause mortality for every 5 kg / m2increase in body mass index (BMI) over 25 kg / m2(BMI ranges: normal < 25 kg / m2, overweight: 25-29.9 kg / m2, and obese > 30 kg / m2). In Canada alone, the proportion of adults overweight and obese was 61.9 % (26.1 % obese) in 2015 and 64.7 % (30 % obese) in 2022 (Statistics Canada, 2023). In the USA, the obesity rate increased from 30.5 % in 1999-2000 to 41.9 % in 2017-2020 (CDC, USA, 2023). Europe is not an exception in that the proportion of adults overweight and obese in Germany recently reached 53.5 % and 19 %, respectively (GEDA 2019 / 2020-EHI). This is indeed a world-wide phenomena, especially considering that eight of the top ten obesity rates are in the Middle East and North Africa (MENA, average 33 % obesity incidence). Thus, it is estimated that more than half of the world’s population will be overweight or obese within 12 years unless treatment and prevention strategies are improved, with predicted global costs exceeding $ 4 trillion in 2035 (World Obesity Federation, 2023).

[0003] Obesity is caused by an imbalance between caloric intake vs. caloric expenditure; thus, treatment strategies broadly focus on either decreasing energy intake, increasing energy output, or a combination of both. These strategies typically include lifestyle management (exercise, healthy food choices, and caloric restriction such as very low calorie diets (VLCD)) or more intensive solutions, such as weight-loss drugs, anti-diabetes medications, and / or bariatric surgery, when justified. An individualized approach that combines several therapies (polytherapy) is almost always necessary to sustain adherence, minimize relapses / bodyweight regain, and achieve clinically meaningful results in the long-term. Importantly, both fat mass and lean mass are independent and strong predictors of quality of life (QOL) and mortality risk.Thus, an ideal weight management program should aim to simultaneously reduce fat mass and increase lean mass (or at least maintain); thus, enhancing the overall body composition index (BCI; lean body mass (LBM) / fat mass) by increasing LBM and lowering fat mass.

[0004] Obesity is linked to a toxic metabolic environment (e.g., dysglycemia, dyslipidemia, and systemic inflammation) that drives intracellular pathogenesis of other peripheral tissues, such as the liver and muscles (i.e., ectopic lipid deposition, oxidative damage, mitochondrial dysfunction, inflammasome activation, and impaired insulin signaling / insulin resistance), and ultimately limits treatment efficacy and benefits. For example, obesity and aging are both independently associated with "anabolic resistance", which is defined as a blunted growth response by the muscles to one or more anabolic factors, such as exercise (mainly str ength / resi stance training), dietary modifications (i.e., protein intake), and / or hormonal therapy. As such, the concept of anabolic resistance is proven in both obesity and aging, and while the underlying pathogenic mechanisms may be slightly different between conditions, a strategy that is able to simultaneously improve muscle growth, fat loss, and the body composition index, would be highly desirable in the treatment of both aging and metabolic disease states (e.g., obesity, type 2 diabetes, metabolic syndrome, etc.). .

[0005] In general, successful weight loss strategies include a dietary component (i.e., reduced caloric intake; less nutrient-dense, processed, and high-fat / sugar foods) and a physical activity component (i.e., increased caloric expenditure; endurance and / or strength / resistance exercise) that are individualized and sustainable in the long term. In fact, in essentially every weight loss drug study published to date, all participants are told to try to lower energy intake by 500 kcal / d and to exercise at least 150 min / week. As an example in a recent study of the antiobesity drug tirzepatide, the instructions to all participants, “.. .included regular lifestyle counselling sessions, delivered by a dietitian or a qualified health care professional, to help the participants adhere to healthful, balanced meals, with a deficit of 500 calories per day, and at least 150 minutes of physical activity per week” (Jastreboff, A., A Engl J Med 2022;387:205- 216). Unfortunately, only 20% of obese individuals can preserve and stabilize the weight loss in the long-term, and more than half regain most of their weight loss within a year. Furthermore, one major drawback of traditional weight-loss programs is that -20-40% of the total weight-loss comes from lean body mass (LBM, for example, skeletal muscle, connective tissue, and viscera),with the remainder being fat mass. Thus, individuals engaging in traditional weight management, such as caloric restriction, typically exhibit poor muscle quality as well as reduced muscle mass following significant weight loss. Although the loss of fat mass can exceed the loss of muscle mass to yield a body composition index (BCI, LBM / fat mass) that appears to be better after antiobesity drugs, the aim is the preservation or increase in LBM concurrent with body fat loss. Consequently, a strategy that would allow for the maintenance or improvement of lean body mass during body fat loss (such as exercise training, VLCD, bariatric surgery or pharmacological interventions) would be desirable.

[0006] When traditional life-style based weight management strategies fail, various weight loss drugs, anti-diabetic medications, and / or bariatric surgery may be used for treating metabolic comorbidities and obesity. Recent studies using drugs that were initially developed for the treatment of diabetes have initiated a massive interest in their role as anti-obesity medications including; glucagon-like peptide-1 receptor agonists (GLP-1RA, such as semaglutide (e.g., Ozempic and Wegovy), dual glucose-dependent insulinotropic polypeptide and GLP-1 receptor agonists (GIP / GLP-lRA)(e.g., tirzepatide (e.g., Mounjaro and Zepbound)), and sodium-glucose cotransporter-2 (SGLT2 inhibitors (SGLT2i, e.g., dapagliflozin, and canagliflozin). However, these treatments are not risk-free. For example, GLP-1RA, GIP / GLP-1RA, and SGLT2i drugs are consistently associated with a loss of LBM (Sargeant, J., Endocrinology and Metabolism, 2019;34:247262). Bariatric surgery is also associated with significant LBM / muscle loss, and a meta-analysis did not find a significant increase in LBM even with exercise training (Bellicha, A., Obesity Reviews, 2021;22, S4). Excessive post-operative muscle deterioration is detrimental to strength, function and QOL of patients, especially in older adults. Interestingly, a reduction of energy intake by 500 - 700 kcal / d led to a loss of LBM in spite of the participants consuming 1 g / kg / d of high quality protein + calcium + vitamin D + exercise training (endurance, resistance or both)(Villareal, D., N. Engl. J. Med. 217; 376: 1943). Another study showed that higher protein (> 1 g / kg / d), calcium and vitamin D attenuated, but did not eliminate, the reduction of LBM seen after 6 months of moderate caloric deficit (- 500 kcal / d)(Ogilvie, A, Obesity (Silver Spring), 2022:30: 1411). Thus, most current weight management strategies induce simultaneous loss of body fat and LBM / muscle mass even with protein intakes > 1.0 g / kg / d + calcium + vitamin D + exercise (endurance, resistance or both combined). Consequently, it would be desirable to have a strategy that can attenuate the loss of LBM / muscle with any weight lossstrategy, particularly when using more severe weight loss solutions, such as pharmacological or surgical options.

[0007] One approach for minimizing the loss of muscle quality and / or quantity during weight-loss is high-protein / low-carbohydrate diets. However, carbohydrate-restriction is difficult to sustain and muscle wasting may still be evident with increased protein intake. Moreover, the anabolic effects of both dietary proteins and standard protein supplements, such as whey concentrates or isolates (e.g., fast-acting, high-quality milk proteins), and meat have been shown to be blunted in obesity (Beals et al. Am J Clin Nutr. 2016. 104: 1014-1022; Smeunix, B., et al., 2017. J Clin Endocrin Metab. 102:3535-3545) and aging (Moore et al. J Gerontol A Biol Sci MedSci. 2015 Jan;70(l):57-62).

[0008] The most effective strategy to increase muscle mass and strength in younger and older adults of all body types is resistance exercise training. Studies have shown that participation in a resistance exercise program at least twice per week can lead to an increase in muscle mass and strength in younger and older adults, albeit with an attenuated response in the very old (> 85 y). Resistance exercise programs generally do not result in a lowering of body fat percentage in men or women, regardless of BMI; however, body fat is at least preserved in most studies. The main dietary strategy to increase / optimize the muscle mass gains induced by resistance exercise is to have a higher protein intake, especially in older adults where they show an anabolic resistance to the stimulatory effects of protein supplementation. The consensus statement from the protein for aging (PROT-AGE) committee was that older adults (> 65 y) should consume 1.0 - 1.2 gPRO / kg / d and > 1.2 gPRO / kg / d if they are engaging in resistance or endurance exercise (Bauer, J., et al., 2013, J Am Diet Assoc. 14:542-559).

[0009] It has been shown that being overweight or obese confers a form of “anabolic resistance” to the stimulatory effects of both protein (amino acids) and / or resistance exercise up lean body mass (LBM, mainly muscle). This concept was first demonstrated in rats where it was found that obese rats had a markedly reduced mixed muscle and myofibrillar fraction protein synthetic response to resistance exercise vs lean rats (Nilsson, M., et al., FASEBJ. 2013.27:3905-3916). Another study found that the acute resistance exercise stimulation of the protein synthetic response for both whole muscle and the myofibrillar fraction was the same in rats fed ahigh-fat vs normal fat diet (Ato, et al., 2021. J Appl Physiol. 131 :442-453). An obesity-induced anabolic resistance was noted in older obese inactive individuals in response to 15 g of milk protein isolate with muscle protein synthesis rates increasing more in the lean (+ 38 %) vs. obese (+ 9 %) following ingestion at rest (Smeunix, B., et al., 2017. J Clin Endocr in Metab. 102:3535- 3545, 2017). Another group found that the muscle protein synthetic response to 170 g of pork (~36 g PRO) was blunted in obese vs normal weight young adults (Beals, J., et al., 2016, Am J Clin Nutr ACPAC M Cli Finally, a study in younger adults found no difference in the acute increase in mixed muscle protein synthetic rates between lean and obese subjects (Huston, C., 2018, Physiol Reports. 6(14): el3799). In a review paper summarizing all of the studies evaluating the effect of obesity-induced resistance to protein / amino acids and / or exercise, the only conclusion was that obese people should ensure that they get at least 1.2 gPRO / kg / d (Beals, J, et al., 2019, Front Nutr. 6; 87).

[0010] Obesity also appears to lead to a state of resistance to the loss of body fat. For example, as shown by Samjoo et al. (Nutr and Diabetes, 2013, 3:e88), lean men lost 3.9 % body fat percentage whilst obese men only lost 0.6 % body fat percentage after 3 months of endurance exercise training. It was also found that while lean women did not lose body fat, obese women gained body fat (49 increasted to > 50 %) following 3 months of endurance exercise training (Devries, M, et. al., Free Radical Biology and Exercise., 2008, 45:502-511).

[0011] Collectively, the combinatory state of muscle / LBM anabolic resistance and resistance to the loss of body fat may be referred to as body composition resistance (BCR). Specifically, BCR refers to both the anabolic resistance to the usual stimulatory effects of protein / amino acid intake and / or exercise upon lean body / muscle mass and the relative resistance to the loss (or actual gain) of body fat in response to interventions designed to both increase lean body mass and / or lower body fat. Ideally, strategies to treat obesity should have a favorable effect on both components of the body composition index (BCI), namely overcoming BCR to lead to an reduction in body fat with a maintenance or ideally, an increase in LBMT

[0012] Due to the almost ubiquitous inability of overweight and obese humans to simultaneously lose body fat and simultaneously maintain or increase LBM / muscle mass, it would be desirable to provide a method of treating body composition resistance in mammals in need thereof, for example, in human and / or non-human mammals who are overweight or obese,aging, or who have metabolic disease, and who may optionally partake in body composition optimization strategies to increase LBM and / or lower % body fat such as exercise, VLCD, bariatric surgery or the use of anti-obesity drugs.Summary of the Invention

[0013] A method of treating body composition resistance (BCR) has now been developed which is based on the use of a multi -nutrient composition comprising the following nutrients: protein, creatine, vitamin D, calcium, and optionally omega-3 fatty acids. The composition has surprisingly been found to be useful for treatment in mammals who have body composition resistance, including but not limited to those who are aging, obese, or those who have metabolic disease.

[0014] Thus, in a first aspect of the present invention, a method of treating body composition resistance in a mammal is provided comprising administering to the mammal for a sufficient period of time the following nutrients: protein, creatine, vitamin D, calcium, and optionally omega-3 fatty acids.

[0015] In another aspect, a method of treating body composition resistance in a mammal is provided comprising administering to the mammal protein in an amount of 10- 100g, creatine in an amount of 1 -5g, vitamin D in an amount of 50-4000 IU, calcium in an amount of 100-1000 mg, and omega-3 fatty acids in an amount of 500-15,000 mg.

[0016] In embodiments, the method of treating BCR in a body composition resistant mammal resulted in an increase in lean body mass, muscle mass, muscle strength, function and muscle quality, a loss of body fat, and / or an increase in body composition index (LBM / fat mass).

[0017] In embodiments, the body composition resistant mammal treated by a method of the invention is a mammal that is overweight or obese, aging, has metabolic disease, is performing regular resistance and / or endurance exercise, is taking a GLP-1 receptor agonist, a GIP receptor agonist or an SGLT2 inhibitor (SGLT2i), is on a low or very low calorie diet (e.g. high-protein / low-carbohydrate diet), and / or has undergone bariatric surgery.

[0018] These and other aspects of the present invention will become apparent in the detailed description that follows, by reference to the following figures.Brief Description of the Figures

[0019] Figure 1 graphically illustrates the increase in total lean mass (A) and appendicular skeletal muscle mass (B) in non-obese vs obese older adults undergoing three months of home-based resistance exercise training (HBRE) + step count advice resulting from daily supplementation of a multi-ingredient composition in accordance with an embodiment (M5, black bars) vs. standard treatment (PL, collagen protein, white bars).

[0020] Figure 2 graphically illustrates the benefits of daily supplementation of the multiingredient composition (M5) on total fat mass (A) and body fat percentage (B) in non-obese vs obese older adults undergoing three months of HBRE.

[0021] Figure 3 graphically illustrates the benefits of daily supplementation of the multiingredient composition (M5) on body composition (A; body composition index; BCI) and a measure of sarcopenia risk and mortality risk (B; appendicular skeletal muscle mass index; ASMI) in non-obese vs obese older adults undergoing three months of HBRE.

[0022] Figure 4 graphically illustrates the benefits of daily supplementation of the multiingredient composition (M5) on maximal strength, including isometric knee extension (A), and leg press (B), in non-obese vs obese older adults undergoing three months of HBRE.

[0023] Figure 5 graphically illustrates the benefits of daily supplementation of the multiingredient composition (M5) on overall muscle quality index in non-obese vs the obese older adults undergoing three months of HBRE.

[0024] Figure 6 graphically illustrates the benefits of daily supplementation of the multiingredient composition (M5) on functional testing and performance, including A) 5 x Sit-to- Stand test, B) 4-Step Stair Climb test, and C) Get Up and Go test in non-obese vs obese older adults undergoing three months of HBRE.

[0025] Figure 7 graphically illustrates the effects of daily intake of the multi-ingredient composition (M5) on systemic drivers of anabolic resistance, body fat gain, and muscle loss in non-obese vs. obese older adults, including: A) dysglycemia, B) systemic inflammation, and C) dyslipidemia. Panel 4 provides a visual diagram of the pathogenic cascade that is mitigated bydaily intake of the multi-ingredient composition (M5) vs. standard treatment (collagen at 1.4 gPRO / kg / d + HBRE / step advice).

[0026] Figure 8 graphically summarizes the overall adaptations in lean mass (A), strength (B), performance (C), and anabolic response (D) to daily intake of M5 vs placebo (PL A; collagen) in the obese subgroup (obese / Mets), specifically, following three months of HBRE.

[0027] Figure 9 graphically illustrates the clinical benefits of daily supplementation of the multi-ingredient composition (M5) vs placebo (PLA; collagen) on sarcopenic obesity risk in the obese subgroup (obese / MetS), specifically, following three months of HBRE.

[0028] Figure 10 graphically illustrates the increase in total lean mass (A) and appendicular skeletal muscle mass (B) in overweight and obese males and females (18-45 years of age) undergoing three months of supervised endurance and resistance training (EnduRX) resulting from daily supplementation of the multi-ingredient composition (M5) and antioxidants in accordance with an embodiment (M5 / T7, black bars) vs. standard treatment (PLA, collagen protein + microcrystalline cellulose, white bars).

[0029] Figure 11 graphically illustrates the benefits of daily supplementation of the multi-ingredient composition (M5) + an antioxidant weight loss supplement (T7, combined = M5 / T7) on total fat mass (A) and body fat percentage (B) in overweight and obese men and women (18-45 years of age) undergoing three months of supervised endurance and resistance training (EnduRX).

[0030] Figure 12 graphically illustrates the benefits of daily supplementation of the multi ingredient composition + a weight loss supplement (M5 / T7) on body re-composition and lean-tofat mass ratios: A) body composition index; TLMZFM; B) TLM / % body fat; C) ASM / FM; and D) ASM / % body fat; in overweight and obese males and females (18-45 years of age) undergoing three months of supervised endurance and resistance training (EnduRX).

[0031] Figure 13 graphically illustrates the benefits of daily supplementation of the multi ingredient composition + antioxidants (M5 / T7) on a negative predictor of mortality risk (ASM / H2; ASMI) in overweight and obese males and females (18-45 years of age) undergoing three months of supervised endurance and resistance training (EnduRX).

[0032] Figure 14 graphically illustrates that muscle mass is improved during semaglutide treatment in Western diet-fed (WD) C57BL6 / J mice by daily supplementation of the multiingredient composition in accordance with an embodiment (WD+M5, yellow bars) vs. standard treatment (WD, white bars).

[0033] Figure 15 graphically illustrates that muscle / fat ratio is improved by daily intake of the multi-ingredient composition (M5) vs standard diet (WD) during semaglutide treatment in Western diet-fed C57BL6 / J mice.Detailed Description of the Invention

[0034] A method of treating body composition resistance in a mammal in need thereof is provided comprising administering to the mammal the following nutrients: protein, creatine, vitamin D, calcium and optionally an n-3 fatty acid. The nutrients may be administered in one or more compositions.

[0035] The term “anabolic resistance” refers herein to reduced or blunted stimulation of muscle protein synthesis in response to one or more anabolic factors, such as exercise (e.g. strength training), dietary modifications (i.e. protein intake), and / or hormonal therapy, which generally results in an increase in skeletal muscle mass. Anabolic resistance may result from one or more of aging, obesity, dysglycemia, dyslipidemia and / or inflammation.

[0036] The term “body composition resistance” refers herein to the combination of anabolic resistance (leading to a loss of LBM) and resistance to the loss of body fat and / or to the preservation of body composition index in response to strategies designed to lower body fat incuding; exercise, a lower carbohydrate and higher protein intake (> 1.2 gPRO / kg / d), bariatric surgery, obesity drugs, and very low calorie diets. Body composition resistance may result, for example, from one or more of aging, obesity, dysglycemia, dyslipidemia and / or inflammation. Thus, a body composition resistant mammal is a mammal that is obese or aging, has metabolic disease, is performing regular resistance and / or endurance exercise, is taking a GLP-1 receptor agonist, a GIP receptor agonist or an SGLT2 inhibitor (SGLT2i), is on a very low calorie diet (e.g. high-protein / low-carbohydrate diet), and / or has undergone bariatric surgery.

[0037] The terms “treat’ ’ / ’treating” and “treatment” refer to the prevention, cure, amelioration, or the slowing of the progression of body composition resistance.

[0038] The present method comprises administration of at least one protein source. The protein source may be selected from any suitable protein source, including an animal source, a dairy source, a plant source, an insect source or any combination thereof. Non-limiting examples of insect sources of protein include: cricket protein, grasshopper protein, mealworm protein, earthworm protein and any combination thereof. Non-limiting examples of animal sources of protein include: cow protein, pig protein, goat protein, lamb protein, poultry protein (such as chicken, duck, goose, pheasant and the like), wild game protein, seafood protein (such as fish and shellfish) and any combination thereof. Non-limiting examples of dairy sources of protein include: whey protein, whey protein concentrate, whey protein isolate, milk protein concentrate, milk protein isolate, powdered fat-free milk, micellar casein, acid casein, potassium caseinate, calcium caseinate, sodium caseinate and any combination thereof. Non-limiting examples of plant sources of protein include: pea protein, yeast protein, soy protein, corn protein, wheat protein, rice protein, canola protein, peanut protein, bean protein, lentil protein, and any combination thereof. The protein source may be non-hydrolyzed, partially hydrolyzed or hydrolyzed and may be in the form of an intact protein, amino acid or peptide. Non-limiting examples of amino acids may include essential amino acids such as: leucine, isoleucine, valine, tryptophan, methionine, threonine, phenylalanine and lysine and semi-essential amino acids such as: histidine and arginine and non-essential amino acids such as tyrosine, aspartic acid, glycine, alanine, cysteine, arginine, glutamic acid, proline, glutamine, serine, asparagine, taurine and any combination thereof. Preferably, the protein source is a high-quality protein source, at least comprising each of the essential amino acids. Preferably, the protein source is whey protein isolate optionally enriched with leucine which has been found to enhance muscle protein synthesis. In one embodiment, the protein source of a multi -nutrient composition for admininistration in accordance with an embodiment comprises about 1-95% of the dry weight of the multi -nutrient composition, such as from about 20-60%, or from about 30-50% of the dry weight of the composition. In another embodiment, the multi -nutrient composition comprises about l-100g of a protein source, for example, up to about 10 g, 20 g, 30 g, 40 g, 50 g, 60 g, 70 g, 80 g or 90 g. In particular embodiments, the composition comprises about 10- 100g, 15-85g or 30-50g. The term “about” is used herein to refer to an amount within 10% or less of theindicated amount, and preferably an amount within 5% or less, wherein the amount is either greater or less than the indicated amount.

[0039] In one embodiment, the protein source comprises a combination of protein sources, such as a fast-acting or fast release protein source, and a slow-acting or slower release protein source. “Fast release” protein refers to protein having a rapid absorption rate into the blood stream on ingestion, for example, about 10 grams per hour. Examples of fast release proteins include protein having a high leucine content, e.g. 8-10% leucine content, such as whey, whey isolates and whey hydrolysates. “Slow release” protein refers to protein having a slower rate of absorption into the blood stream on ingestion, for example, 5-10 grams per 4-5 hours. Examples of slow release protein include casein, egg protein, milk, and pea protein.

[0040] Leucine has been determined to combine with a protein source, such as whey protein isolate, to provide a synergistic effect. Thus, a reduced amount of the protein source is required when combined with leucine to yield a given muscle protein synthesis response than the amount of protein required if administered without leucine. Thus, leucine enrichment of a protein may reduce the total amount of protein in the composition by about 10%, 20%, 30%, 40% or more, while still achieving a desirable muscle protein synthesis response. For example, when a suboptimal dose of whey protein isolate (e.g. 6.25 g of whey) is administered in conjunction with 4.25 g of leucine (whey + leu), the muscle protein synthesis response was comparable to that resulting from the administration of 25 g of whey protein (with no added leucine). Thus, in this case, a composition comprising leucine-enriched whey protein totalling only 10.5 g of total protein source yielded a muscle protein synthesis response similar to that achieved with a composition comprising 25 g of whey protein.

[0041] Creatine for use in the present method may be in any suitable form, such as creatine monohydrate, creatine anhydrous, creatine citrate, creatine ethyl ester, creatine nitrate, creatine magnesium chelate, creatine hydrochloride, creatine ascorbate, creatine malate, creatine pyruvate, creatine phosphate, creatine citrate malate, creatine tartrate, creatine HMB (P-hydroxy P- methylbutyrate), effervescent creatine, creatine titrate, buffered creatine, micronized creatine and any combination thereof. Preferably, the creatine is creatine monohydrate. In one embodiment, the creatine of the multi -nutrient composition comprises about 0. l%-40% of the dry weight of themulti -nutrient composition, such as about 1-20%, or about 5-10% of the dry weight of the composition. In another embodiment, the multi -nutrient composition comprises about 0.1-10g of creatine and preferably, about l-5g, such as 3g.

[0042] Vitamin D in any suitable form may be used in the present method, including but not limited to, vitamin D2 (ergocalciferol), vitamin D3 (cholecalciferol) and any combination thereof. Preferably, the vitamin D is vitamin D3. In one embodiment, the vitamin D of the multinutrient composition comprises about 0.0000004-0.001% of the dry weight of a multi -nutrient composition for use in the present method, such as about 0.00001-0.0005%, or about 0.00005- 0.0001% of the dry weight of the composition. In another embodiment, the multi -nutrient composition comprises about 50-4000 IU of vitamin D and preferably, between about 250-3000 IU, such as 500-2000 IU of vitamin D. For clarity, IU refers to International Units. For vitamin D, 1 pg is 40 IU.

[0043] A source of calcium in any suitable form may be used in the present method, such as calcium carbonate, bonemeal calcium, dolomite calcium, calcium citrate, oyster shell calcium, calcium gluconate, calcium lactate, calcium lactobionate, calcium phosphate, calcium citrate malate, calcium orotate, calcium ascorbate, calcium hydroxyapatite, microcrystalline hydroxyapatite and any combination thereof. Preferably, the source of calcium is a calcium salt such as calcium carbonate, or a source of calcium with enhanced bioavailability such as calcium citrate, or a combination thereof. In one embodiment, a multi -nutrient composition for use in the present method comprises a source of calcium in an amount of about 0.1-10% of the dry weight of the multi-nutrient composition, such as about 0.1-5%, or 1-2% of the dry weight of the composition. In another embodiment, the multi-nutrient composition comprises about 10-2000 mg of a calcium source, and preferably between about 100-1500 mg, 150-1000 mg, such as 400 mg of a calcium source.

[0044] N-3 fatty acids for use in the present method are polyunsaturated fatty acids having a double bond between the third and fourth carbons of the carbon chain. N-3 fatty acids are also commonly referred to as omega-3 fatty acids. Non-limiting examples of n-3 fatty acids include eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), alpha-linolenic acid (ALA), stearidonic acid, docosapentaenoic acid, and combinations thereof. The n-3 fatty acid may beobtained from any suitable n-3 fatty acid source, including a cold water fish (e.g. cod, salmon, tuna, sardines, mackerel, krill and squid), algae, dark leafy green vegetables, plant and plant seed oils (e.g. flaxseed oil, canola oil and walnut oil), nuts (e.g. walnuts) and any combination thereof. The n-3 fatty acid may be in the triglyceride form typically found in nature, in ethyl ester form or free-fatty acid form, and may be in liquid or powdered form. Preferably, the n-3 fatty acid is a combination of EPA and DHA. In one embodiment, the amount of n-3 fatty acid for use in the present method comprises about 0. l%-20% of the dry weight of a multi -nutrient composition, such as about 0.1-10%, or about 1-4% of the dry weight of the composition. In another embodiment, the multi -nutrient composition comprises about 100 mg- 15,000 mg of n-3 fatty acid, and preferably, about 500-10,000 mg, or 1000-9000 mg of n-3 fatty acid. In one embodiment, the composition comprises 100-3000 mg each of DHA and EPA.

[0045] In one embodiment, the present method comprises administering to a mammal, for example on a regular basis such as daily, protein in an amount of 10- 100g, creatine in an amount of l-10g, vitamin D in an amount of 500-2000 IU, calcium in an amount of 100-1500 mg, and optionally omega-3 fatty acids in an amount of 500-15,000 mg. In one embodiment, a daily or regular dose of nutrients for use in the present method comprises 10-100g of protein (e.g. 15-60g of whey protein isolate and 5-30g of micellar casein), 1 -5g of creatine monohydrate, 500-2000 IU of vitamin D3, 100-1000 mg of calcium carbonate, and optionally omega-3 fatty acids in an amount of 500-15,000 mg. In another embodiment, a daily or regular dose for use in the present method comprises 20-30g of whey protein isolate, 10-20 g of micellar casein, l-5g of creatine monohydrate, 600-1800 IU of vitamin D3, 200-800 mg of calcium carbonate or citrate, and optionally omega-3 fatty acids such as 100-2000mg of DHA and 500-3000mg of EPA, or 800- lOOOmg of DHA and 1000-2000mg of EPA. The therapeutic effect of the present multinutrients results from the combination thereof across their concentration ranges.

[0046] According to one embodiment, a multi -nutrient composition as described may be used as a sole, primary, or supplemental source of nutrition. The multi -nutrient composition may comprise other essential nutrients required in an adequate diet, such as vitamins, minerals, carbohydrates and fats as would be appreciated by one of skill in the art, particularly if the multinutrient composition is used as a sole source of nutrition.

[0047] The multi -nutrient composition may be formulated with at least one additional source of nutrition, including, but not limited to, carbohydrates, additional lipids, fibre, vitamins, minerals, antioxidants, prebiotics, probiotics, phytochemicals or phytonutrients.

[0048] The multi -nutrient composition may comprise any food-grade source of fibre which is suitable for oral administration to an individual. Suitable sources of fibre include the following non-limiting examples: water-soluble dietary fiber such as beta-glucans, pectin, xylose, plant gums, inulin and alginates, and insoluble dietary fiber such as lignin, beta-glucans, xanthan gum, resistant starches and combinations thereof.

[0049] The multi -nutrient composition may comprise any food-grade source of additional lipids which is suitable for oral administration to an individual. The term “additional lipids” as used herein is intended to include lipids other than n-3 fatty acids. Suitable sources of additional lipids include the following non-limiting examples: olive oil, safflower oil, canola oil, coconut oil, com oil, palm oil, palm kernel oil, soybean oil, peanut oil, fish oil, almond oil, sunflower oil, butter, lard, and sources of medium chain triglycerides, long chain triglycerides, monoglycerides, diglycerides and combinations thereof.

[0050] The multi -nutrient composition may comprise any food-grade source of vitamins and minerals which are suitable for oral administration to an individual. Suitable vitamins include the following non-limiting examples: vitamin A, vitamin C, vitamin E, vitamin K, thiamine, riboflavin, niacin, pantothenic acid, pyridoxine, folic acid, cobalamin, biotin, carotenoids (e.g. lutein, beta-carotene, lycopene and cryptoxanthin), choline, inositol and combinations thereof, and suitable minerals include, but are not limited to, phosphorus, selenium, chromium, zinc, molybdenum, iodine, chloride, phosphorus, manganese, fluoride, potassium, iron, copper, magnesium, sodium and combinations thereof.

[0051] The multi -nutrient composition may comprise any food-grade source of antioxidants which are suitable for oral administration to an individual. Suitable antioxidants include the following non-limiting examples: coenzyme Q10, vitamin A, vitamin E, vitamin C, folic acid, alpha lipoic acid, and beta-carotene, iron, copper, butylated hydroxyamisole, butylated hydroxytoluene, propyl gallate, tertiary butylhydroquinone, resveratrol, and plant phytonutrients or phytochemicals (e.g. flavonoids and lignin). Herbs or herbal extracts (e.g. oregano, Goji berry,dill, garden thyme, rosemary and peppermint), forskolin, tea leaves or tea leaf extracts (e.g. camellia sinensis), coffee bean extracts (e.g. coffea canephora and coffea arabica), brewed coffee or tea or brewed coffee or tea extracts (e.g. green tea, black tea, oolong tea and coffee robusta) and other plants or plant extracts (e.g. beet root) may also be used as a source of antioxidants, as well as combinations of any of the antioxidants. In an embodiment, the administration of components of the multi -nutrient composition together with one or more antioxidants enhances the effect of the composition to mitigate body composition resistance, for example, by promoting lean body mass gain, increasing lean-to-fat mass ratio and increasing strength. In another embodiment, the antioxidants are selected from green coffee bean extract, green tea extract, beet root extract, forskolin, CoQlO, a-lipoic acid, and vitamin E. In another embodiment, the antioxidants comprise green coffee bean extract, green tea extract, beet root extract, forskolin, CoQlO, a-lipoic acid, and vitamin E. In a further embodiment, the antioxidants are administered at a daily dosage of 50- lOOOmg of green tea extract, 50-1000mg of green coffee bean extract, 15mg-100mg of forskolin, 50-5000mg of beetroot extract, 50-900mg of coenzyme Q10, 50mg-900mg alpha lipoic acid and / or 50-900mg of vitamin E. In another embodiment, the antioxidants may be included in a composition in the following amounts: 0.1-5% w / w of green tea extract, 0.1-5% w / w of green coffee bean extract, 0.005-0.05% w / w of forskolin, 0.1-5% w / w of beetroot extract, 0.1-5% w / w of coenzyme Q10, 0.1-5% w / w alpha lipoic acid and 0.1-5% w / w of vitamin E.

[0052] The multi-nutrient composition may comprise any food-grade source of prebiotics which are suitable for oral administration to an individual. Suitable prebiotics include the following non-limiting examples: dietary fibers and carbohydrate polymers such as cellulose, inulin, gums, trans-galactooligosaccharide, fructans, resistant starches, xylooligosaccharides, hemicelluloses, pectin, sugar alcohols, beta-glucans and combinations thereof.

[0053] The multi -nutrient composition may comprise any food-grade source of probiotics which are suitable for oral administration to an individual. Suitable probiotics include the following non-limiting examples: Lactobacillus Acidophilus, Lactobacillus Reuteri, Lactobacillus Rhamnosus, Lactobacillus Gasseri, Lactobacillus Salivarius, Lactobacillus Bulgaricus, Lactobacillus Helventicus, Lactobacillus Silivarus, Lactobacillus Plantarum, Lactobacillus Casei, Lactobacillus Paracassei, Lactobacillus Fermentum, Bifidobacterium Breve, Bifidobacterium Lactis, Bifidobacterium Longum, Bifidobacterium Bifidum, BifidobacteriumInfantis, Bifidobacterium Bifidum, Bacillus Coagulans, Saccharomyces Boulardii, Pediococcus Acidlacti and combinations thereof.

[0054] The multi -nutrient composition may comprise any food-grade source of phytochemicals or phytonutrients which are suitable for oral administration to an individual. Suitable phytochemicals or phytonutrients include the following non-limiting examples: phytosterols including sterols (e.g. cempesterol) and stanol (e.g. sitostanol), soy flavonoids (e.g. genistein and glycitein), garlic and organosulfur compounds (e.g. L-cysteine sulfoxides and y- glutamyl-L-cysteine peptides), carotenoids (e.g. zeaxanthin alpha-carotene, beta-carotene, lycopene, beta-cryptoxanthin and lutein), resveratrol, curcumin, fiber (e.g. lignin and cellulose), indole 3-carbinol and condensation products (e.g. 3,3'-diindolylmethane and 5,11-dihydroindolo- [3,2-b]carbazole), chlorophyll and chlorophyllin isothiocyanates, isothiocyanates (e.g. sulforaphane and benzyl isothiocyanate) and combinations thereof.

[0055] In one embodiment, at least one physiologically acceptable excipient may be included in the multi -nutrient composition. The term “physiologically acceptable” is used herein to refer to excipients which are food-grade and thus, acceptable for consumption or administration to an individual. Examples of suitable excipients, which are not to be construed as limiting, include flavouring agents, sweetening agents, anti-caking agents / flowing agents, emulsifiers, stabilizers, masking agents, colorants, preservatives, disintegrants, binders, thickeners and pH adjusters.

[0056] Non-limiting examples of flavouring agents include natural or artificial flavours such as fruit flavours (e.g. raspberry, orange, apple, pomegranate, mixed berry, lemon, lime, watermelon, strawberry, blueberry, pineapple, coconut, grape, cherry, banana, peach, mango, kiwifruit, cranberry), sodium sources (e.g. sodium chloride and monosodium glutamate), high fructose com syrup, vanilla, chocolate, unsweetened chocolate, honey, molasses, brown sugar, coffee, cocoa, mint, maple, almond, or extracts or combinations thereof. Savoury flavourings may also be used (e.g. beef, chicken or vegetable flavourings).

[0057] Non-limiting examples of sweetening agents include natural sweeteners such as, glucose, fructose, sucrose, dextrose, maltose, brown sugar, molasses, honey, maple syrup, com syrup, high fructose com syrup, erythritol, xylitol, sorbitol, isomalt, monatin, monellin, curculin,brazzein, tagatose and mannitol, and artificial sweeteners such as aspartame, acesulfame K, saccharin cyclamate and sucralose.

[0058] Non-limiting examples of anti-caking agents / fl owing agents include silicates and calcium or magnesium stearates.

[0059] Non-limiting examples of emulsifiers include agar, gums, egg yok, lecithin, monostearate, monosodium phosphate, monoglycerides, diglycerides and alginates.

[0060] Non-limiting examples of stabilizers include glycerine, agar, gums, alginates and pectin.

[0061] Non-limiting examples of masking agents include glycerine, sodium chloride, peppermint, lemon-lime, mint, cherry, black liquorice, peach, apricot, raspberry, or sweetening agents such as aspartame or sucrose.

[0062] Non-limiting examples of colorants include those which are suitable for inclusion in foods. For example, commonly used colorants include FD&C blue #1, FD&C blue #2, FD&C citrus red #2, FD&C green #3, FD&C red #3, FD&C red #40, FD&C yellow #5 and FD&C yellow #6.

[0063] Non-limiting examples of preservatives include butylated hydroxyanisole, butylated hydroxytoluene, ethylenediaminetetraacetic acid, nitrates (e.g. sodium nitrate), sulfites (sodium bisulfite), benzoates (sodium benzoate), sorbates (e.g. sodium sorbate) and sodium chloride.

[0064] Non-limiting examples of disintegrants include starches (e.g. potato starch), alginic acid, cellulose and derivatives thereof, and calcium silicate.

[0065] Non-limiting examples of binders include stearic acid, gelatin, saccharides and derivatives thereof, sugar alcohols, polyethylene glycol and cellulose.

[0066] Non-limiting examples of thickeners include polysaccharide-based thickeners such as vegetable gums, pectin and starches or protein-based thickeners such as gelatin, egg white and collagen.

[0067] Non-limiting examples of pH adjusters include citric acid, ammonium carbonate, ammonium phosphate, calcium carbonate, sodium hydroxide, malic acid and phosphoric acid.

[0068] As will be appreciated by one of skill in the art, for each type of excipient (e.g. flavouring agent, sweetener, emulsifier, preservative, etc.), a single excipient may be used, or a combination of two or more may be used.

[0069] In one embodiment, a multi -nutrient composition for use in the present method may be formulated for oral administration including, for example, in solid, semi-solid, liquid, semiliquid, powder, suspension, emulsion, solution, ready-to-drink beverage, gel, bar, pill, tablet or capsule form. The term “oral” or “orally” as used herein is intended to include any method in which the multi -nutrient composition is introduced into the digestive tract including the stomach and small intestine. Examples of oral administration may include administration via mouth, directly into the stomach using a feeding tube, through the nose to the stomach via a feeding tube and through the nose to the small intestine via a feeding tube. In a preferred embodiment, the multi -nutrient composition is provided as a powder or a bar. The powdered composition may be reconstituted in water or any suitable liquid immediately prior to consumption. When provided in powdered form, the multi -nutrient composition may be packaged in individual use containers, packets or sachets, or in larger bulk containers.

[0070] The multi -nutrient composition may also be formulated for administration parenterally, and may be combined with at least one pharmaceutically acceptable adjuvant. The expression "pharmaceutically acceptable" means acceptable for use in the pharmaceutical and veterinary arts, i.e. not being unacceptably toxic or otherwise unsuitable. Examples of pharmaceutically acceptable adjuvants include diluents, excipients and the like. Reference may be made to "Remington's: The Science and Practice of Pharmacy", 21st Ed., Lippincott Williams & Wilkins, 2005, for guidance on drug formulations generally. The selection of adjuvant depends on the intended mode of administration of the composition. In one embodiment of the invention, the composition is formulated for administration by infusion, or by injection either subcutaneously or intravenously. The composition may be prepared as an aqueous solution in sterile and pyrogen- free form and optionally buffered or made isotonic. Thus, the composition may be administered in distilled water or, more desirably, in saline, phosphate-buffered saline or 5% dextrose solution.The composition may also be formulated for topical administration. Creams, lotions and ointments may be prepared for topical application using an appropriate base such as a triglyceride base. Such creams, lotions and ointments may also contain a surface active agent. Aerosol formulations may also be prepared in which suitable propellant adjuvants are used. Other adjuvants may also be added to the composition regardless of how it is to be administered for example, anti-microbial agents may be added to the composition to prevent microbial growth over prolonged storage periods.

[0071] The composition for administration in any form may include a coating or may be encased in a protective material to prevent undesirable degradation thereof by enzymes, acids or by other conditions that may affect the therapeutic activity thereof. For example, the composition may be formulated as nanoparticles, liposomes, micelles and the like.

[0072] The multi -nutrient composition may be administered in an effective amount to a mammal in need thereof, regularly for a sufficient period of time, which may include one or more times per day, for a period ranging from one day to chronic or long-term administration. Generally, the expression “a sufficient period of time” refers to at least a period of time within which body composition in a mammal has been treated as defined herein. The term “effective amount” as used herein, refers to an amount which achieves the effects desired by the mammal, without surpassing an amount which may cause undesirable side effects. For example, an effective amount of the multi -nutrient composition may be administered 1, 2, 3, 4, 5, 6 or more times per day, or an effective amount of the multi -nutrient composition may be divided into 2, 3, 4, 5, 6 or more servings to be administered throughout the day. In one embodiment, the multi -nutrient composition is administered to a mammal in the morning when first waking up. The multi -nutrient composition may also be administered to a mammal in the morning and then administered again prior to the mammal retiring to bed. The multi -nutrient composition may be administered to a mammal in need thereof for 1, 2, 3, 4, 5, 6 or 7 days in a week and for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more weeks. In one embodiment, the multi -nutrient composition is administered chronically to a mammal in need thereof. The term “chronically” as used herein refers to the administration of the multi -nutrient composition for a period of at least 2-4 or more months, for example, administration of the multi -nutrient composition on a continual basis beyond 6 months, at a frequency of at least 2 days / week, and preferably at least 3 or more days a week.

[0073] The term “mammal” or “individual” is used herein to encompass human and nonhuman mammals, such as cats, dogs, horses, cows, goats, sheep, rodents and the like. A mammal in need of treatment is a mammal who is experiencing body composition resistance, and may include, but is not limited to, obese mammals, aging mammals (e.g. mammals who are experiencing time-related deterioration of normal physiological functions) and mammals with disease such as metabolic disease (dysglycemia, dyslipidemia, and / or elevated inflammation).

[0074] The components of the composition may be administered in conjunction, either together, in a single composition, or individually, at the same time or at different times. For example, the protein source, creatine, vitamin D and calcium may be administered together as a powder, while the n-3 fatty acid source may be administered separately as an oil once during the day at the same time as the powder is administered, or at a different time, and at the same frequency or at different frequencies. For example, the powder may be administered twice daily, while the oil is administered once a day.

[0075] Accordingly, in a further aspect of the invention, a kit is provided comprising a first component comprising a protein, creatine, vitamin D and calcium, and a second component comprising an n-3 fatty acid. The first component comprises a first composition that may be in the form of a powder provided in bulk form or as individual doses, e.g. in packets, sachets or capsules, and the second component comprises a composition that may be in the form of an oil in bulk form or as individual doses, e.g. in capsules.

[0076] The first composition comprises a protein, creatine, vitamin D and calcium. Sources of these ingredients are described in detail herein. The protein, thus, may be an animal, dairy, plant, insect or combination thereof. In one embodiment, the protein is a dairy protein such as a whey protein, whey protein concentrate, whey protein isolate, whey protein hydrolysate, milk protein concentrate, milk protein isolate, powdered fat-free milk, micellar casein, acid casein, potassium caseinate, calcium caseinate, sodium caseinate and any combination thereof. In one embodiment, the protein source comprises a combination of a fast release protein such as a whey protein, concentrate, isolate or hydrolysate, with a slower release protein such as casein and / or other slow release protein sources such as pea protein or egg protein. Creatine in any suitable form may be incorporated such as creatine monohydrate, creatine anhydrous, creatine citrate,creatine ethyl ester, creatine nitrate, creatine magnesium chelate, creatine hydrochloride, creatine ascorbate, creatine malate, creatine pyruvate, creatine phosphate, creatine citrate malate, creatine tartrate, creatine HMB (P-hydroxy P-methylbutyrate), effervescent creatine, creatine titrate, buffered creatine, micronized creatine and any combination thereof. The Vitamin D may be any suitable form, such as vitamin D2 and / or vitamin D3. The calcium may be, for example, calcium carbonate, bonemeal calcium, dolomite calcium, calcium citrate, oyster shell calcium, calcium gluconate, calcium lactate, calcium lactobionate, calcium phosphate, calcium citrate malate, calcium orotate, calcium ascorbate, calcium hydroxyapatite, microcrystalline hydroxyapatite and any combination thereof. The amount of the selected protein(s), creatine, vitamin D and calcium in the first composition is as specified herein.

[0077] The second composition comprises at least one n-3 fatty acid (omega-3 fatty acids). Examples of suitable n-3 fatty acids include EP A, DHA, ALA, stearidonic acid, docosapentaenoic acid, and combinations thereof. Preferably, the n-3 fatty acid is a combination of EPA and DHA. The amount of the selected n-3 fatty acid(s) in the second composition is as specified herein.

[0078] The first and second compositions may additionally include at least one physiological acceptable excipient, e.g. an excipient that does not impact the function of the first and and second compositions. Examples of suitable excipients are known in the art, and include, for example, flavouring agents, sweetening agents, anti-caking agents, flowing agents, emulsifiers, stabilizers, masking agents, colorants, preservatives, disintegrants, binders, thickeners, pH adjusters, carbohydrate and oil.

[0079] In the present treatment of body composition resistance in a mammal, while not wishing to be limited by any particular theory or mode of action, the method of administering the selected multi -nutrient composition or composition is useful to treat body composition resistance by targeting and reducing key drivers of anabolic resistance such as one or more of dysglycemia, inflammation and / or dyslipidemia. In particular, the present method of administering the multinutrient composition has surprisingly been shown to be effective to prevent increases in biomarker levels associated with these drivers of anabolic resistance as compared to protein-only treated controls. For example, the present treatment is shown to prevent increases in inflammation (C- reactive protein) of more than 5%, and preferably, to prevent increases of more than 2%, 1%, orto result in no increase in inflammation (i.e. 0%), or results in reduced biomarker levels of dysglycemia and dyslipidemia, in a population of obese individuals as compared to protein-only treated controls or untreated controls. In one embodiment, the present method results in significantly reduced levels of HgbAlc as a biomarker of dysglycemia, and total cholesterol, low density lipoprotein (LDL) and / or triglycerides, as biomarkers of dyslipidemia, as compared to protein-only treated controls in obese populations.

[0080] The present method also exhibits an increase in LBM, muscle mass, an increase in muscle strength, function and muscle quality, a loss of body fat, and / or an increase in body composition index (LBM / fat mass) in a mammal. The method comprises administering to the individual an effective amount of the multi -nutrient composition. The term “improve” is used herein) to refer to either a healthy increase in the property, i.e. an increase in the property (e.g. LBM, muscle mass, muscle strength, muscle quality and BCI), or a decrease in a property (e.g. fat mass) that is considered to promote health / functional capacity. Thus, the present method is applicable to mammals that wish to improve one or more of; lean mass, fat mass or body fat, muscle mass, muscle quality, muscle strength, and / or body composition index (LBM / fat mass), who may be performing exercise and / or other strategies / conditions that promote body fat loss including; very low calorie diets, starvation, therapies such as GLP-1 + / - GIP receptor agonists or SGLT2 inhibitors, multi-ingredient weight loss supplements, or bariatric surgery.

[0081] As used herein, the term “lean body mass (LBM)” refers to the fat-free and bone- free mass of an individual (i.e. the total body mass of an individual minus the mass contributed by fat tissue and bone tissue). Lean body mass primarily reflects the total amount of skeletal muscle mass in an individual. Lean body mass may be measured as whole body LBM, or the LBM of specific regions of the body such as appendicular LBM, lower body LBM or upper body LBM using X-ray technology, e.g. dual energy X-ray absorptiometry (DEXA). The body composition index (BCI) is derived from the LBM / fat mass (both in kg) with higher numbers reflecting more lean mass or muscle and / or less body fat in an individual. Improved LBM / BCI refers to an increase in the amount of LBM in an individual by at least about 0.5%, and preferably an increase of about 1% or more, e.g. by 5%, 10%, 30%, 50%, 70% or greater to the LBM / BCI of the individual prior to treatment with the present composition, or a reduction in the rate of LBM / BCI loss by at least about 1% or more, and preferably a reduction of the rate of LBM / BCI loss by about 5% or more,e.g. by 10%, 30%, 50%, 70%, 90% or greater of the rate of LBM / BCI loss of the individual prior to treatment with the present composition.

[0082] As used herein, the term “body fat” refers to the total fat mass of an individual or to the fat mass of specific regions of the body. During healthy weight loss, the majority of body weight loss is derived from the reduction of excess fat stores in an individual. Improved body fat is a reduction of body fat in an individual by at least about 0.5%, and preferably a reduction of about 1% or more, e.g. by 5%, 10%, 30%, 50%, 70% or greater of body fat in the individual prior to treatment with the present composition, or a reduction in the rate of rising body fat by at least about 1% or more, and preferably a reduction of about 5% or more, e.g. by 10%, 30%, 50%, 70%, 90% or greater to the rate of body fat increase prior to treatment.

[0083] As used herein, the term “muscle” refers to skeletal muscles and non-skeletal muscle. Such muscle may be consciously controlled such as the diaphragm muscles. The term “muscle strength” as used herein, refers to the amount of force which may be generated by a muscle or group of muscles. Muscle strength may be measured by numerous methods, such as a 1- repetition max (1-RM) test, a grip strength test or a callisthenic-type exercise test (e.g. push-ups). Improved muscle mass and / or muscle strength refers to an increase in muscle mass or an increase in muscle strength in an individual by at least about 0.5%, and preferably an increase of about 1% or more, e.g. by 5%, 10%, 30%, 50%, 70% or greater from the muscle mass or muscle strength of the individual prior to treatment with the present composition, or at least a reduction in the rate of muscle mass or muscle strength loss in an individual by at least about 1% or more, and preferably a reduction in the rate of muscle mass or muscle strength loss by about 5% or more, e.g. by 10%, 30%, 50%, 70%, 90% or greater from the rate of muscle strength loss in the individual prior to treatment with the present composition.

[0084] Muscle quality index (MQI) is used herein to refer to the actual quality of muscle. MQI is calculated by dividing appendicular muscle strength by muscle mass, and defines the strength of the muscle per unit of muscle mass. Improved MQI is an increase in the quality of the muscle of an individual by at least about 0.5%, and preferably about 1% or more, e.g. by 5%, 10%, 30%, 50%, 70% or greater as compared to the MQI of the individual prior to treatment in accordance with the present method, or at least a reduction in the rate of loss of MQI by at leastabout 1% or more, and preferably a reduction in the rate of loss of MQI of an individual by about 5% or more, e.g. by 10%, 30%, 50%, 70%, 90% or greater as compared to the MQI of the individual prior to treatment with the present composition.

[0085] As used herein, the term “muscle fiber size” refers to the size of any of the 3 major skeletal muscle fiber types in humans including, type I, type 2a and type 2x. Size reflects the mean fiber area or diameter of the respective fiber type. Improved muscle fiber size is an increase in the size of the muscle fibers of an individual by at least about 0.5%, and preferably about 1% or more, e.g. by 5%, 10%, 30%, 50%, 70% or greater to the muscle fiber size prior to treatment in accordance with the present method, or at least a reduction in the rate of loss of muscle fiber size by at least about 1% or more, and preferably a reduction in the rate of loss of muscle fiber size by about 5% or more, e.g. by 10%, 30%, 50%, 70%, 90% or greater to the muscle fiber size prior to treatment with the present composition.

[0086] As used herein, the term “functional capacity” refers to the ability to complete timed and / or measured tasks designed to reflect the ability to perform activities of daily living. Such tasks include, but are not restricted to; six minute walk test (6MWT, distance in m), timed up and go (TUG, in seconds), 5 time sit-stand time (seconds), 4,6,10 m walk test (seconds), 4-stair climb (seconds), and Short Physical Performance Battery (SPPB). Improved functional capacity refers to an increase in the functional ability of an individual by at least about 0.5%, and preferably an increase of about 1% or more, e.g. by 5%, 10%, 30%, 50%, 70% or greater of an aspect of functional capacity of the individual prior to treatment with the present composition, or at least a reduction in the rate of functional capacity loss by at least about 1% or more, and preferably a reduction of the rate of functional capacity loss by about 5% or more, e.g. by 10%, 30%, 50%, 70%, 90% or greater of the rate of functional capacity loss of the individual prior to treatment with the present composition.

[0087] As used herein, the term “low or very low calorie diet” refers to the intentional reduction in energy intake by an individual for the purpose of weight loss. In contrast, the reduction in energy intake from a low or very low calorie diet due to illness or other circumstances is termed, “starvation”. Very low calorie diets are generally under 800 kcal / d, for example, 400 - 500 kcal / d for women, and 600 - 700 kcal / d for men. In starvation conditions the energy intakecan range from 0 kcal / d for short periods of time to < 60 % of daily intake for longer periods of time.

[0088] As used herein, the term “weight loss supplements” refers to either a single or a multi-ingredient supplement that may comprise one or more of: alpha lipoic acid, green tea extract, green coffee bean extract, forskolin, conjugated linoleic acid, caffeine, bitter melon, and the like.

[0089] As used herein, the term “bariatric surgery” refers to a surgical procedure designed to reduce the ability to ingest energy by lowering the capacity of the stomach (for example, via a gastric sleeve, stapling or other method to reduce the size of the stomach), or by bypassing the stomach (e.g. using any method of diverting food from the esophagus to the intestine directly).

[0090] In another embodiment, a method is provided for increasing lean mass, muscle strength, and dysglycemina, and reducing systemic inflammation levels, blood cholesterol levels and blood triglyceride levels in an individual, comprising administering to the individual the present multi -nutrient composition.

[0091] In a further embodiment, the present method is administered to a mammal who is performing exercise. The term “exercise” is meant to encompass endurance exercise, high- intensity interval training, resistance exercise, and the like, e.g. exercise that achieves a level of working of at least about 3-6 metabolic equivalents (METS), and combinations thereof (e.g. any combination of endurance exercise, high-intensity interval exercise, or > 50 % of the one repetition maximum (resistance exercise)). METS is the energy expenditure of a physical activity or exercise defined as the ratio of the metabolic rate of an exercising individual (and therefore the rate of energy consumption) during a specific physical activity to a reference basal metabolic rate. In a preferred embodiment, the exercise is performed on a regular basis. Regularly performing exercise refers to the performance of exercise for a duration of at least a month and preferably chronically such as for 2, 4 or 6 or more months, at a frequency of at least 2 days / week, and preferably at least 3 or more days a week, for a period of at least 30 consecutive minutes per day, preferably 45 minutes or greater, such as 60 minutes or greater, or 75 - 90 minutes or more. Exercise may include endurance activities such as brisk walking, jogging, running, dancing, swimming, bicycling, sports, interval training, resistance exercise, and the like. Interval training refers to repetitive bouts of exercise that may be at high or lower intensity provided it meets minimal METS requirements.High intensity interval training would include activities such as sprints (e.g. 10 second to 4 minute sprints) followed by a recovery time (e.g. of 10 seconds to 4 minutes). The term “resistance exercise” refers to weight training or other resistance exercise (plyometrics, hydraulic machines, etc.) with a resistance of at least 50% of the one repetition maximum, performed in sets of repetitions (for example, 8-15 repetitions), followed by a recovery between sets, for a period of time sufficient to achieve minimal METS requirements. One repetition maximum is the maximal voluntary contraction strength for a single movement where a second movement is impossible.

[0092] A multi -nutrient composition in accordance with the present method may be administered to a mammal being treated at any time relative to the performance of exercise, i.e. before, during or following the exercise, or any combination thereof. In one embodiment, the multi -nutrient composition is administered to the mammal immediately following exercise.

[0093] The present invention advantageously provides a method of treating body composition resistance in mammals in need thereof, such as mammals who are aging, obese and / or mammals with metabolic disease. The selected multi -nutrient combination unexpectedly targets drivers of body composition resistance, muscle loss and obesity, and results in an efficacy that is significantly enhanced in comparison to previously recommended exercise and protein-only (> 1.2 g protein / kg / d) treatments. It is particularly efficaceous for use to treat obese individuals in combination with exercise training (traditional weight management strategy) or for those attempting weight-loss with bariatric surgery or drug therapy, such as GLP-1 receptor agonists (RA), GIP / GLP-1RA, and SGLT2i’s. A synergistic outcome of the present method is that it simultaneously induces an increase of LBM / muscle mass or an attenuation of LBM / muscle mass loss during weight loss, an improvement in muscle function and / or strength, an improvement of body composition index (BCI), and / or a loss of fat mass / % body fat, whilst treating a mammal with body composition resistance.

[0094] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present application herein described for which they are suitable as would be understood by a person skilled in the art.EXAMPLES

[0095] The following non-limiting examples are illustrative of the present application. While the present application has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.

[0096] All references referred to herein are incorporated by reference in their entirety to the same extent as if each reference was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.Example 1 - A novel multi-ingredient supplement improves the root causes of anabolic resistance in obese adults and promotes simultaneous muscle growth, body fat loss, and enhanced body composition index (BCD.

[0097] To determine if a multi-ingredient composition (M5), in accordance with an embodiment of the invention, comprising a fast-acting anabolic agent (whey protein isolate), slow-acting anabolic agent (casein protein isolate), an anabolic and metabolic support agent (creatine monohydrate), n-3 PUFAs (EPA / DHA)), and muscle / bone-supporting vitamins and minerals (vitamin D3 and calcium), could:1) improve underlying etiological factors in body composition resistance (e.g., inflammation, dysglycemia, and dyslipidemia);2) simultaneously enhance muscle growth (or preserve / attenuate loss) and body fat loss (e.g., body fat mass / % body fat) and improve body composition index (BCI);3) increase maximal strength; and4) improve muscle function and performance in obese older adults undergoing three months of home-based resistance exercise (FIBRE) + step count advice.

[0098] A randomized, double-blind, and placebo-controlled clinical trial was conducted in which 32 participants were randomly allocated into placebo (PLA)) or multi-ingredient supplement (MIS; M5) groups, and analyzed within non-obese (n = 21) and obese (n = 11) body mass index classifications retrospectively. All subjects underwent daily supplement / placebo intake and 3 d / wk home-based resistance exercise (HBRE; whole-body elastic band training) for a total of 12 weeks. Finally, participants were instructed to walk atleast 5000 steps on exercise days and 10,000 steps on rest days.

[0099] One daily serving of the multi-ingredient supplement (M5) contained whey protein isolate (24 g / d), micellar casein (16 g / d), creatine monohydrate (3 g / d), vitamin D3 (1000 lU / d), calcium (416 mg / d), and omega-3 containing fish-oil (EP A; 1.51 g / d, DHA; 0.95 g / d). Sucrose, stevia, and chocolate flavor were included as flavoring agents in both PLA (i.e., collagen and sunflower oil) and M5, which were isocaloric (272 kcal per serving / day), protein-matched (40 g protein / day), and identical in flavor, smell, and appearance (Table 1).Table 1. Ingredients and energy content per serving in ‘Muscle 5’ and Placebo*Note that the vitamin E was identical between groups and was a taste masker / m vitro anti-oxidant and not at a dose designed for in vivo anti-oxidant effects.

[0100] The HBRE program was performed on three non-consecutive days per week and consisted of six lower body and six upper body elastic band exercises, including biceps curl, triceps extension, lateral raise, seated row, bench press, abdominal crunch, calf raise, chair squat, knee extension, knee flexion, hip flexion, and dorsi flexion. In accordance with the progressive overload principle, the participants were encouraged to increase the intensity of the exercise (i.e. elastic band resistance) throughout the course of the study. They were also instructed to walk at least 5000 steps on exercise days and 10,000 steps on rest days.

[0101] Co-primary outcomes were pre-to-post improvements in body composition by dual X-ray absorptiometry (DXA), including total lean mass (TLM), appendicular skeletal muscle mass (ASM), appendicular skeletal muscle mass index (ASMI; ASM / H2), fat mass, body fat percentage, and body composition index (BCI; lean mass / fat mass ratio).Secondary outcomes were pre-to-post improvements in overall muscle quality (MQ), muscle function / performance (5 x Sit-to-Stand, Timed Up and Go and 4-Step Stair Climb times, and Short Physical Performance Battery (SPPB)), and maximal strength (1-RM leg press and isometric knee extension). Other important outcomes of this study were the driving systemic factors of anabolic resistance, body fat gain and muscle loss in aging and obese conditions, including inflammation, dysgycemia, and dyslipidemia.

[0102] For all pre- and post- clinical assessments, subjects were in the fasted state in the morning. All participants underwent muscle strength and functional testing, body composition scans, and venous blood draws on both visits. Exercise, diet, and supplement logs were provided to keep track of activity levels, macronutrient intake and supplementation / HBRE adherence. Step counts and dietary intakes (3-day food record) were assessed one week prior to and following the interventional period.

[0103] All methods and procedures in this randomized , double-blind, placebo- controlled trial were approved by the Hamilton Integrated Research Ethics Board (2018- 46S6- GRA). Study participants were informed about the potential risks of participation prior to giving their informed consent. This clinical trial was registered at clinicaltrials.gov (NCT03S36871).Method / Results

[0104] To evaluate changes in basic anthropometry and macronutrient intakes, bodyweight (BW), body mass index (BMI), energy, protein, carbohydrate, and fat intakes were assessed pre and post intervention in all participants (Table 2). These results indicate that there were no significant changes in bodyweights or BMIs in either O-PLA or O-M5 groups in non- obese or obese cohorts. Self-reported energy and macronutrient intakes were largely the same between O-PLA and O-M5 groups, while total caloric intakes appeared lower in O-M5 groups post intervention. As expected, daily protein intakes were significantly increased in both O-PLA and O-M5 groups, clearly exceeding the recommended dietary allowance of 0.8 g / kg BW / day. The majority of participants also exceeded 1.2 g protein / kg BW / day, including all placebo groups, which is the recommended protein intake by experts in the field of protein and aging.Table 2. Participant descriptives and self-reported energy and macronutrient intakes#Post study energy and macronutrient intakes include both diet sources and the isocaloric placebo or MIS.

[0105] To assess if daily supplementation with the multi-ingredient supplement (MIS; O- M5) enhanced muscle gains over placebo (O- PLA; collagen), percent changes in total lean mass (TLM) (Fig 1 A) and appendicular muscle mass (ASM) (Fig IB) were analyzed by DXA in all participants pre-to-post intervention (% A pre-post). These findings demonstrate that the multiingredient composition enhanced total lean mass and appendicular muscle mass gains over calorie-and protein-matched placebo; thus, effectively mitigating anabolic resistance in non- obese (BMI 20-30) and obese older adults. However, surprisingly, muscle gain in non-obese individuals (BMI 20-30) was shown to be less than 2%, while muscle gain in obese individuals was shown to be at least about 3% (from muscle loss of about 2% in the placebo group to musclegain of about 1% in the M5 treated group). A similar effect was found in ASM, with an increase in ASM in non-obese individuals of less than 1% between the placebo and M5 groups, while the increase in ASM in obese individuals was shown to be at least about 5% (from muscle loss of more than 4% in the placebo group to muscle gain of more than 1% in the M5 treated group).

[0106] To assess if daily supplementation with the multi -ingredient composition (O-M5) reduced body fat gain over placebo (O-PLA), change in total fat mass (Fig 2A) and body fat percentage (Fig 2B) were assessed by DXA pre and post intervention in all participants. These results demonstrate that the multi-ingredient composition promoted total body fat loss and lowered percentage body fat in both non-obese and obese older adults vs. calorie- and protein- matched placebo.

[0107] To assess if daily supplementation of the multi-ingredient composition (M5) improved overall body composition, sarcopenia risk and mortality risk over calorie- and protein- matched placebo, the body composition index (Fig 3 A; BCI) and the appendicular skeletal muscle mass index (Fig 3B; ASMI) were assessed by DXA pre and post intervention in all participants. These findings demonstrate that the multi-ingredient composition (M5) increased both BCI and ASMI over placebo / standard treatment; thus, improving overall body composition and mitigating sarcopenia risk and mortality risk in non-obese and obese older adults. However, surprisingly, the improvement resulting in obese individuals in both BCI and ASMI was notably superior than the improvement seen in non-obese individuals. For example, ASMI increased by about 1% between the non-obese PLA and M5 groups, but increased by greater than 5% between the obese PLA and M5 groups.

[0108] To assess if daily supplementation of the multi-ingredient composition (M5) improved maximal strength gains over calorie- and protein-matched placebo, maximal strength was assessed by isometric knee extension (Fig 4A) and leg press (Fig 4B) pre and post intervention in all participants. These findings demonstrate that the multi-ingredient composition (M5) enhanced maximal strength gains over placebo / standard treatment in non-obese and obese older adults. Again, the results were surprisingly enhanced in the obese population with an improvement in isometric knee extension between PLA and M5 groups of almost double in obese individuals as compared to non-obese individuals, and an improvement in leg pressstrength of about 30% in obese individuals as compared to about 10% in non-obese individuals.

[0109] To assess if daily supplementation of the multi-ingredient composition improved overall muscle quality vs. calorie- and protein-matched placebo, the muscle quality index (MQ) was determined in all participants (Fig 5). These findings demonstrate that the multi-ingredient composition (M5) enhanced the overall muscle quality index (MQ) over placebo / standard treatment in obese older adults. The -12% improvement of MQ in obese adults was notably better than the non-obese groups as well.

[0110] To assess if daily supplementation of the multi-ingredient composition enhanced muscle function and performance over calorie- and protein-matched placebo, 5 x Sit-to-Stand (Fig 6A), 4-Step Stair Climb (Fig 6B) and Get Up and Go (Fig 6C) times were measured pre and post intervention in all participants. These results demonstrate that the multi-ingredient composition (M5) improved muscle function and performance (e.g., decreased execution times) over placebo / standard treatment in obese older adults, with little or no improvement seen in non- obese individuals.

[0111] To determine if daily intake of the multi-ingredient composition (M5, black bars) indeed attenuated the root causes of anabolic resistance vs. calorie- and protein-matched placebo (collagen PL A, white bars), systemic markers of dysglycemia (Panel 1, Fig 7 A), inflammation (Panel 2, Fig 7B), and dyslipidemia (Panel 3, Fig 7C) were assessed pre- and post- intervention in non-obese vs obese participants. These findings clearly demonstrate that the multi-ingredient composition (M5) mitigated the root causes / pathogenic cascade (Panel 4) of body composition resistance (BCR) that drive body fat gain and muscle wasting over standard treatment / placebo in obese older adults.

[0112] To assess the overall adaptive response (A%) to the multimodal intervention in the participants at highest risk for sarcopenic obesity, a subgroup analysis was performed in overweight (BMI > 27) and obese (BMI > 30) individuals with at least one other risk factor for metabolic syndrome (including, baseline obesity (> 27 BMI, > 28% body fat, and > 0.9 m waist- to-hip ratio), dyslipidemia (> 1.69 mmol / L triglycerides and < 0.9 mmol / L HDL), dysglycemia (> 5.7% HbAlc), hypertension (> 140 systolic and > 90 diastolic), as well as systemic inflammation (> 3 mg / L CRP)) (obese / MetS; PLA n = 8, M5, n =12). Each participant was ranked from 1 (lowest responder) to 20 (highest responder) for lean mass (LM, allometric LM,and LM / BF ratios), strength (isometric knee extension and leg press) and performance (4SSC and SPPB score). An overall rank of the anabolic response could then be generated from the average ranks of A lean mass (Fig. 8A; +80% M5 vs. PLA ), A strength (Fig. 8B; +43% M5 vs. PLA), and A performance (Fig. 8C; +43% M5 vs. PLA). For each area of assessment (i.e., Figs. 8A-C), the M5 group consistently ranked higher than the PLA group, indicating a more robust anabolic response in the M5 group (Fig. 8D; +45% M5 vs. PLA). This finding is novel and clearly demonstrates that a lower-quality protein source (PLA; collagen) is inferior to a higher- quality, multi-ingredient supplement (M5) for mitigating age- and obesity -associated anabolic resistance in older adults.

[0113] Finally, to assess if the adaptive response to the multimodal intervention affected polymorbidity in the obese / MetS subgroup, a Sarcopenic Obesity Risk Rank was generated for all participants. Thus, individuals were ranked according to baseline obesity (% BF; low-to-high), allometric lean mass (ASM / BW; high-to-low), 5XSTS time (low-to-high), and number of metabolic risk factors (low-to-high), with a ranking of 1 representing the lowest and a ranking of 20 representing the highest risk for each diagnostic criteria, respectively, which were then averaged to obtain an overall Sarcopenic Obesity Risk Rank. The M5 group exhibited a significant decrease in the risk ranking vs PLA following three months of HBRE (see Fig. 9; M5 -7% vs. PLA +31%) indicating that the differential adaptive response to the multimodal intervention led to a significant decrease in sarcopenic obesity risk. This finding is the most striking piece of evidence that demonstrates that the high-quality, multi-ingredient supplement (M5) reduces sarcopenic obesity risk in older adults.

[0114] Surprisingly, it was found that the present multi-ingredient supplement (M5) containing a higher-quality protein source (whey / casein), creatine, vitamin D, calcium and n-3 fatty acids, led to greater improvements in lean body mass, strength, function, and body composition index (LBMZFM) concurrent with reductions in body fat as compared to an isoenergetic, collagen supplement that was matched for total protein content (40 g / day). Thus, the present multi -nutrient composition led to superior adaptations and body re-composition in older adults (both non-obese and obese) even when calories and total protein content were matched between products. In fact, the total protein intake (diet and supplement) for the collagen group (PLA) was higher (1.4 gPRO / kg / d) than for the M5 group (1.1 gPRO / kg / d) during the intervention.Despite this, the adaptations were significantly greater in the M5 group, including superior improvements in the underlying drivers of body composition / anabolic resistance in the obese participants (M5), while these benefits were not observed in those consuming the isocaloric collagen supplement (PLA).

[0115] Unexpectedly, the obese PLA group even exhibited lowered LBM, ASM and body composition index (LBM / fat mass), increased body fat, showed worsening to minimal functional capacity improvements, and minimal improvement (knee extension) to worsening (leg press) in strength following the training program despite consuming 1.4 gPRO / kg / d per day (i.e. which is above expert recommendations). In contrast, it was striking that age-matched obese men performing the same exercise program showed robust increases in strength, muscle mass, body composition index, functional capacity with simultaneous reductions in body fat using the novel multi-ingredient composition (M5), despite consuming less total protein per day (1.2 gPRO / kg / d) as compared to the 1.4 gPRO / kg / d in the PLA group.

[0116] Collectively, these data show superiority of the present multi-ingredient composition (M5) over the standard treatment / placebo (exercise and total daily protein intake > 1.2 gPRO / kg / d) to mitigate body composition resistance by increasing LBM, leg strength, muscle quality, and functional capacity concurrent with lowering body fat mass and percentage in obese adults. Specifically, it is shown that the present multi-ingredient composition (M5) attenuates the driving pathogenic mechanisms of body composition resistance, and therefore, allows for simultaneous muscle gain and fat loss in obese individuals, which is a novel and unexpected finding given that the standard treatment for those with obesity and aging does not improve nor attenuate the loss of LBM / muscle mass.Example 2 - A combination of multi-ingredient supplement (M5) and antioxidants (T7) promote muscle growth, body fat loss, and body re-composition in overweight and obese males and females

[0118] To determine if the multi -ingredient composition (M5), in accordance with an embodiment of the invention, comprising a fast-acting anabolic agent (whey protein isolate), slow-acting anabolic agent (casein protein isolate), an anabolic and metabolic support agent (creatine monohydrate), muscle / bone-supporting vitamins and minerals (vitamin D3 and calcium), could promote a more favorable BCI in overweight and obese men and women whileexercising and taking an antioxidant supplement (T7; green coffee bean extract, green tea extract, beet root extract, forskolin, CoQlO, a-lipoic acid, and vitamin E) having weight loss properties as described in WO 2019 / 178689, the contents of which are incorporated herein by reference. The specific outcomes were assessed using:1. muscle growth (TLM and ASM),2. body fat loss (e.g., body fat mass / % body fat), and3. the body composition index (BCI; lean mass / fat mass ratio) in overweight and obese individuals undergoing three months of supervised exercise training (EnduRX).

[0119] A randomized, double-blind, and placebo-controlled clinical trial was conducted in which 51 overweight and obese males and females (18-45 years old) were randomly allocated into placebo (PLA) or multi-ingredient supplement (MIS; M5 / T7) groups and underwent daily supplement / placebo intake and 3 d / wk mixed endurance and resistance exercise (EnduRX) for 12 weeks. The participants were instructed to maintain their normal dietary intake and step counts throughout the trial to investigate the effects of MIS and EnduRX on body composition in isolation of other confounding variables.

[0120] One daily serving of the multi-ingredient supplement (M5 / T7) contained whey protein isolate (24 g / d), micellar casein (16 g / d), creatine monohydrate (3 g / d), vitamin D3 (1000 lU / d), calcium (416 mg / d), green coffee bean extract (500 mg / d), green tea extract (500 mg / d), beet root extract (500 mg / d), forskolin (50 mg / d), CoQlO (200 mg / d), a-lipoic acid (400 mg / d), and vitamin E (200 lU / d). Sucrose, stevia, and chocolate flavor were included as flavoring agents in both PLA (i.e., collagen and microcrystalline cellulose) and M5 / T7, which were isocaloric (272 kcal per serving / day), protein-matched (40 g protein / day), and identical in flavor, smell, and appearance (see Table 1 in Example 1).

[0121] The EnduRX program was performed on three days per week and consisted of supervised aerobic exercise (treadmill, elliptical, stationary bike, recumbent bike, or aerobics ), whole-body resistance exercise (3 sets x 20-25 repetitions per exercise; leg press, chest press, hamstring curls, lateral -to-front shoulder raises, knee extension, lat pulldown, squat-to-calf raise, biceps curls, seated Russian twist, and triceps extension), and whole-body elastic band training (3 sets x 20-25 repetitions per exercise; exercises are described in Example 1). Inaccordance with the progressive overload principle, the participants increased the resistance / intensity after 4 and 8 weeks of training. Once weekly, participants engaged in continuous aerobic exercise (30 min / session) and progressed from -55% to -65% VO2peak from the beginning to the end of the trial, respectively. Alternatively, participants were allowed to engage in fartlek training (30 min 50-80% VO2peak) on this training day. Lastly, high- intensity interval training (HIIT; 10 x 1 min at 90% HRmax) and aerobics (moderate intensity) were performed on the remaining two training days.

[0122] Co-primary outcomes were pre-to-post improvements in body composition by dual X-ray absorptiometry (DXA), including total lean mass (TLM), appendicular skeletal muscle mass (ASM), appendicular skeletal muscle mass index (ASMI; ASM / H2), fat mass, body fat percentage, and body composition index (BCI; lean mass / fat mass ratio).

[0123] For all pre- and post- clinical assessments, participants arrived in the rested and fasted state to the laboratory. Assessments included strength, aerobic capacity, anthropometric, and body composition testing on both visits. All participants completed 3-day dietary records pre- and pos-t intervention to determine changes in energy and macronutrient intakes. Habitual physical activity levels were estimated by recording 7-day step counts with pedometers pre- and post- intervention.

[0124] All methods and procedures were approved by the Office of Research Ethics at University of Waterloo (approval number 43396). Study participants were informed about the potential risks of participation prior to giving their informed consent. This clinical trial was registered at clinicaltrials.gov (NCT05384431).Method / Results

[0125] To evaluate changes in basic anthropometry and dietary intakes from daily M5 / T7 or PLA supplementation and EnduRX, bodyweight (BW), body mass index (BMI), energy, protein, carbohydrate, and fat intakes were assessed pre- and post- intervention (Table 3). Both groups exhibited mild reductions in bodyweights (~ -1%) and BMIs, with no significant differences between treatments. Interestingly, the individuals receiving M5 / T7 consumed less calories pre-to-post intervention (from diet), although this clearly did not affectthe anthropometrical outcomes of this study. However, because the whey and casein in M5 (60:40 ratio; humanized milk ratio) are outstanding functional components of milk that have co-evolved to optimize satiety and growth, this in combination with the other components of the composition contributed to the observed body re-composition in the M5 / T7 group (see body composition results). Adherence to the EnduRX and MIS intervention was -90% in both groups, and total daily protein intake (diet + supplement) was -1.2 g protein / kg BW / day during the intervention.Table 3. Example 2: Anthropometry and dietary intakes (food only).Groups Age Bodyweight BMI Energy Protein Protein Carbohydrate Fat(n size) (years) (kg) (kg / in2) (kcal / d) (g / d) (g / kg (g / d) (g / d)BW / d)Pre Pre Post Pre Post Pre Post Pre Post Pre Post Pre Post Pre PostPLA 30 94.9 93 9 32.9 32.6 1994 1871 99 88 1.12 0.96 212 199 84 83(n = 24)M5 / T7 30 97.7 97.2 33.6 33.4 2019 1593 79 72 0.85 0.77 238 180 82 62(n = 27)NOTE: Daily dietary intakes do not include the energy or macronutrients provided by daily intake of PLA (40 g collagen and 272 kcal) or M5 / T7 (40 g whey / casein and 272 kcal).

[0126] To assess if daily supplementation with the multi -ingredient supplement M5 / T7 enhanced muscle gains over placebo (PLA), percent changes in total lean mass (TLM) (Fig. 10A) and appendicular muscle mass (ASM) (Fig. 10B) were analyzed by DXA in all participants pre-to-post intervention (% A pre-post). These findings demonstrate that the multiingredient composition enhanced total lean mass and appendicular muscle mass gains over calorie-and protein-matched placebo; thus, effectively mitigating anabolic resistance in overweight and obese men and women (BMIs > 25). Surprisingly, the pre-to-post improvements in TLM and ASM were not significant in the placebo group, indicating that increasing daily protein intake to > 1.2 g PRO / kg BW / day with a lower quality protein source (collagen) may not be sufficient for improving lean mass during exercise / nutrition polytherapy in overweight or obese individuals.

[0127] To assess if daily supplementation with the multi -ingredient composition M5 / T7 reduced body fat gain over placebo (PLA), change in total fat mass (Fig. 11 A) and body fat percentage (Fig. 1 IB) were assessed by DXA pre- and post- intervention in allparticipants. These results demonstrate that the multi-ingredient composition promoted total body fat loss and lowered percentage body fat in overweight and obese males and females vs. calorie- and protein matched placebo. Importantly, this proves that the M5 / T7 formulation promotes simultaneous LBM gain and body fat loss.

[0128] To assess if daily supplementation of the multi -ingredient composition (M5 / T7) improved overall body composition vs placebo (PLA), lean-to-fat mass ratios were assessed by DXA pre- and post- intervention in all participants, including TLM / FM (body composition index; Fig. 12 A), TLM / %BF (Fig. 12B), ASM / FM (Fig. 12C), and ASM / %BF (Fig. 12D). These results demonstrate that the multi-ingredient composition (M5 / T7) increased all lean-to-fat mass ratios over placebo / standard treatment; thus, improving overall body composition in overweight and obese males and females. These findings also confirm that M5 / T7 promoted body re-composition even when bodyweight or BMI were not drastically changed. Surprisingly, the pre-to-post improvements were significantly inferior in the placebo group, indicating that increasing daily protein intake to > 1.2 g PRO / kg BW / day with a lower quality protein source (collagen) may be insufficient to induce significant changes in body composition in overweight or obese individuals undergoing standard therapy.

[0129] To assess if daily supplementation of the multi -ingredient composition (M5 / T7) improved a negative predictor of mortality risk over calorie- and protein matched placebo, the appendicular skeletal muscle mass index (Fig 13; ASMI) was assessed by DXA pre- and post- intervention. These findings demonstrate that the multi-ingredient composition (M5 / T7) increased ASMI over placebo / standard treatment; thus, reducing mortality risk in overweight and obese males and females.

[0130] To assess if daily supplementation of the multi -ingredient composition (M5 / T7) improved gains in maximal upper and lower body strength and aerobic capacity (VO2peak) vs. placebo, 1 -repetition maximal strength (1-RM; shoulder press and leg press) and VO2peak were assessed pre and post intervention (Table 4). These findings demonstrate that both groups improved maximal strength and VO2peak significantly, while improvements in maximal strength were more robust in the M5 / T7 group. Notably, the addition of antioxidants to the M5 composition did not limit functional adaptations to exercise / nutrition polytherapy in overweight and obese individuals.Table 4. Example 2: Pre-to-post intervention changes in strength and aerobic capacityGroups Shoulder press A Leg press A VO2max A (n size) (1-RM) (1-RM) (mls / kg / min) Pre Post Pre Post Pre PostPLA 83 106 28% 321 522 63% 25.2 29.1 15%(n = 24) M5 / T7 88 120 36% 302 598 98% 24.8 28.0 13%(n = 27)

[0131] Collectively, these data show superiority of the present multi -ingredient composition (M5 / T7) over placebo (collagen) to treat body composition resistance by promoting simultaneous LBM gain, body re-composition (lean-to-fat mass ratios), and strength in overweight and obese males and females undergoing exercise / nutrition polytherapy. Again, a novel and unexpected finding is that increasing protein intake with a lower quality protein source (collagen) is not sufficient for promoting significant body re-composition in this population. Collectively, the addition of antioxidants to the novel M5 formulation effectively mitigates anabolic resistance in obese males and females (18-45 years of age). Further, the present M5 formulation effectively mitigated anabolic resistance in obese males and females (18-45 years of age) when they were also consuming an antioxidant supplement (T7) that lowered body fat (i.e., subjects are able to not just preserve muscle / LBM but actually gain muscle mass / LBM whilst simultaneously taking an antioxidant supplement).Example 3 - A multi-ingredient supplement (M5) improves muscle mass during GLP-1 receptor agonist treatment in Western diet-fed C57BL6 / J mice.

[0132] To determine if the present multi-ingredient composition (M5), in accordance with an embodiment of the invention could improve or maintain muscle mass during treatment with a pharmacological weight loss agent known to effectively treat obesity (GLPl-Ra drug, semaglutide). The specific outcomes used to determine of the composition could attenuate the loss of muscle mass were:1. muscle mass and2. muscle / fat ratio in Western diet-fed (WD) C57BL6 / J mice undergoing semaglutide treatment.

[0133] Western diet-fed male C57BL6 / J mice (WD; n = 24) were matched by body weight and divided into two groups (WD; n = 12 and WD+M5; n = 12). Thereafter, all animals received daily semaglutide injections for six weeks during which the WD mice received a standard high-fat / sucrose diet and the WD+M5 mice a high-protein, isocaloric diet containing the M5 formulation.

[0134] Dietary compositions were as follows for the WD and WD+M5 diets, respectively: energy content (4.6 vs 4.6 kcal / g), protein content (15% vs 23.7% total kcal), carbohydrate content (43.3% vs 34.7% total kcal), and fat content (41.7% vs 41.6% total kcal). Thus, protein content was increased in WD+M5 by -60% by adding whey / casein at 60 / 40 ratio (“humanized milk ratio). In addition, creatine was added (10 g / kg), calcium (1%) and vitamin D3 (2000 lU / kg) doubled, and 40 g lard replaced by Omega-3 s compared to the WD diet.Because the diets were isocaloric, food intakes did not differ significantly between the groups during the intervention.

[0135] At the conclusion of the study, all mice were sacrificed under anesthesia and fast- and slow-twitch muscles of the hindlimb excised, weighed and muscle / fat ratios assessed by a trained technician blinded to the treatment allocations.

[0136] All methods and procedures were approved by the Animal Care Committee (ACC) at York University.Method / Results

[0137] To assess if daily intake of the multi-ingredient formulation M5 in a population on a weight-loss drug improved muscle mass over the standard diet, fast- and slow- twitch muscle groups were carefully removed, cleaned, and weighed on a high-precision balance (Fig 14A-F). These findings demonstrate that the multi-ingredient composition improved muscle mass with -20% over the standard diet; thus, significantly attenuating lean mass loss during semaglutide treatment in Western diet-fed male C57BL6 / J mice.

[0138] To assess if daily intake of the multi-ingredient formulation M5 improved overall body composition during treatment with a weight loss drug, muscle / fat ratios were also assessed (Fig 15). These findings show that the multi -ingredient composition improvedmuscle mass by about -20% over the standard diet; thus, significantly attenuating lean mass loss during semaglutide treatment in Western diet-fed male C57BL6 / J mice.

[0139] Collectively, these data show that using the multi-ingredient composition (M5) promotes LBM gain during pharmacological treatment with weight loss drugs, such as GLP-1 receptor agonists.

Claims

CLAIMS1. A method of treating body composition resistance in a mammal comprising administering to the mammal nutrients comprising protein in an amount of 10- 100g, creatine in an amount of l-10g, vitamin D in an amount of 50-4000 IU, calcium in an amount of 100-1500 mg, and optionally omega-3 fatty acids in an amount of 500-15,000 mg.

2. The method of claim 1, wherein the protein comprises a fast-acting protein and a slow-acting protein.

3. The method of claim 1, wherein the protein comprises whey protein and casein.

4. The method of claim 3, wherein the whey protein is a whey protein isolate, whey protein concentrate or hydrolyzed whey protein.

5. The method of any one of claims 1-4, comprising administration of omega-3 fatty acids comprising eicosapentaenoic acid (EP A) and docosahexaenoic acid (DHA).

6. The method of claim 5, wherein DHA is administered in an amount of 100-2000mg and EPA is administered in an amount of 100-3000mg.

7. The method of claim 5, comprising administering protein in an amount of 10-40 of whey protein isolate, 10-30 g of casein, 1 -5g of creatine monohydrate, 800-1800 IU of vitamin D3, 200- 800 mg of calcium carbonate or citrate, 800-1600mg of DHA and 1000-2500mg of EPA.

8. The method of any one of claims 1-7, wherein said nutrients are in the form of 1 or more compositions.

9. The method of claim 8, wherein the nutrients are administered in the form of a first composition comprising the protein, creatine, vitamin D and calcium and a second composition comprising the DHA and EPA.

10. The method of claim 9, wherein the first composition is in powder form and the second composition is an oil.

11. The method of any one of claims 1-10, wherein the mammal is aging, obese or has metabolic disease.

12. The method of any one of claims 1-10, wherein the mammal is performing regular exercise.

13. The method of any one of claims 1-12, which additionally results in increases in at least one of lean mass, muscle mass, muscle strength, or body composition index, or a decrease in body fat or fat mass, in the mammal as compared to a protein-only treated control.

14. The method of claim 11, wherein the mammal is obese.

15. The method of any one of claims 1-14, which treats dysglycemia as shown by a reduction in the level of HbAlc in the mammal as compared to a protein -only treated control.

16. The method of any one of claims 1-14, which reduces inflammation as shown by a reduction in the level of c-reactive protein in the mammal as compared to a protein-only treated control.

17. The method of any one of claims 1-14, which treats dyslipidemia as shown by a reduction in the level of cholesterol, LDL and / or triglycerides in the mammal as compared to a protein-only treated control.

18. The method of any one of claims 1-17, wherein the mammal is taking a weight loss medication.

19. The method of any one of claims 1-18, wherein one or more antioxidants are administered to the mammal.

20. The method of claim 19, wherein the antioxidants are selected from the group of green coffee bean extract, green tea extract, beet root extract, forskolin, CoQlO, a-lipoic acid, and vitamin E.

21. A method of treating body composition resistance in a mammal that is obese, aging and / or has metabolic disease, the method comprising administering to the mammal for a sufficient period of time, protein in an amount of 10-100g, creatine in an amount of l-5g, vitamin D in an amount of 50- 4000 IU, calcium in an amount of 100-1000 mg, and omega-3 fatty acids in an amount of 500-15,000 mg.

22. The method of claim 21, which results in increases in at least one of lean mass, muscle mass, muscle strength, or body composition index, or a decrease in body fat or fat mass, in the mammal as compared to a protein-only treated control.

23. The method of claim 22, wherein at least one of body composition index and muscle quality index exhibits an increase of at least 5% in comparison to a protein-only treated control.

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