Use of amino acid supplementation to improve muscle protein synthesis

Abstract

To provide a method for stimulating muscle protein synthesis.SOLUTION: A method comprises administering to a subject a composition comprising the following concentrations of amino acids: about 1 to 2% of histidine, about 9 to 11% of isoleucine, about 35 to 38% of leucine, about 14 to 17% of lysine, about 2 to 4% of methionine, about 5 to 7% of phenylalanine, about 8 to 9% of threonine, about 9 to 11% of valine, about 0.005 to 0.8% of tryptophan, and about 8 to 11% of arginine, so that stimulation of muscle protein synthesis is increased compared to a subject to whom the amino acid composition is not administered.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Application No. 62 / 580,86I, filed on November 2, 2017, the entire disclosure of which is incorporated herein by reference.

[0002] Government Rights This invention was made with government support under AR052293 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0003] The present disclosure generally relates to the use of anabolic amino acid compositions for stimulating muscle protein synthesis. Specifically, it discloses compositions and methods of using the same for preventing and / or treating a decrease in muscle mass, muscle strength, muscle function, and physical function, or any one of their combinations, in mammals, particularly adult mammals.

Background Art

[0004] Stimulating muscle protein synthesis requires a large intake of dietary protein. Nutrient assimilation is mainly driven by transferring and incorporating amino acids obtained from dietary protein sources into skeletal muscle proteins. The purpose of this action is to compensate for muscle proteins lost during fasting (post - absorptive) periods, for example, by amino acid oxidation and / or carbon donation for hepatic gluconeogenesis. However, as humans age, the body's efficiency in digesting protein decreases. The decline in muscle mass begins at age 30 and decreases at a rate of 3 - 8% every 10 years, accelerating from age 60. In the elderly over 70 years old, such a decline can reach up to 35 - 40%, so sarcopenia is particularly prominent in the elderly. As a result, the progressive decline in muscle mass associated with aging increases the risk of injury and physical disability.

[0005] Thus, there is a need for dietary supplements that promote the rate of muscle protein synthesis more than can be expected from the consumption of dietary protein.

Summary of the Invention

[0006] Various aspects of the present disclosure include methods and compositions for stimulating muscle protein synthesis. Generally, the present disclosure provides a nutritional composition comprising an effective amount of amino acids that stimulates muscle protein synthesis in a subject as compared to a subject not administered the composition.

[0007] One aspect of the present disclosure is directed to providing a composition comprising histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine, tryptophan, and arginine. In one aspect, the present disclosure provides a composition comprising amino acids at the following concentrations in terms of w / w%: about 1-2% histidine, about 9-11% isoleucine, about 35-38% leucine, about 14-17% lysine, about 2-4% methionine, about 5-7% phenylalanine, about 8-9% threonine, about 9-11% valine, about 0.005-0.8% tryptophan, and about 8-11% arginine.

[0008] In another aspect, muscle protein synthesis is improved by about 50% at a minimum dose of 3.6 grams. In yet another aspect, the composition comprises amino acids at the following concentrations in terms of w / w%: 1.5% histidine, 9.7% isoleucine, 36.4% leucine, 15.2% lysine, 3% methionine, 6.1% phenylalanine, 8.5% threonine, 10% valine, 0.6% tryptophan, and 9% arginine.

[0009] A further aspect of the present disclosure is directed to a method of administering a nutritional composition comprising amino acids at a dose of 12 g once a day, 3.6 g three times a day, 3.6 g once a day, 3.6 g twice a day, 6.7 g once a day, 6.7 g twice a day, or 6.7 g three times a day.

[0010] Another aspect provides a composition that stimulates muscle protein synthesis by more than 50% at a dosage of less than 12 grams, regardless of the presence or absence of exercise, and contains amino acids such as histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine, tryptophan, and arginine.

[0011] In yet another aspect, the present disclosure provides a composition and method of use thereof for stimulating muscle protein synthesis using a dosage of less than 12 grams. In some embodiments, the composition contains amino acids at the following concentrations, converted to w / w%: about 1-2% histidine, about 9-11% isoleucine, about 35-38% leucine, about 14-17% lysine, about 2-4% methionine, about 5-7% phenylalanine, about 8-9% threonine, about 9-11% valine, about 0.005-0.8% tryptophan, and about 8-11% arginine.

[0012] Although multiple embodiments are disclosed, further other embodiments of the present invention will be apparent to those skilled in the art from the following detailed description that illustrates and describes exemplary embodiments of the present invention. Therefore, this detailed description should be considered to be illustrative in nature and not restrictive.

Mode for Carrying Out the Invention

[0013] The present disclosure generally provides methods and compositions comprising certain amino acid formulations that can stimulate the rate of muscle protein synthesis. The Applicants have surprisingly found that the compositions described herein stimulate muscle protein synthesis in an efficient manner. Specifically, the compositions disclosed herein stimulate the production of more muscle protein than the total weight of the components ingested. According to the present invention, it has been discovered that formulations containing free essential amino acids (EAAs) and arginine in specific amounts are very suitable for stimulating muscle protein synthesis. Specifically, it has been found that nutritional compositions containing a mixture of essential amino acids and arginine containing a substantial amount of leucine effectively stimulate muscle protein synthesis. Advantageously, the formulations have been found to be effective in adults in whom the stimulation of muscle protein synthesis is inefficient. Muscle protein synthesis is improved compared to subjects not administered the amino acid composition, and this occurs regardless of the presence or absence of exercise.

[0014] The components used in the preparation of the disclosed compositions, as well as the compositions themselves used within the scope of the methods disclosed herein, are disclosed. Although such substances and other substances are disclosed herein, if combinations, subsets, interactions, groups, etc. of such substances are disclosed, even if specific references to each of the various individual and collective combinations and permutations of these compounds are not explicitly disclosed, each is clearly contemplated and understood to be described herein. For example, if a particular compound is disclosed and considered, and some of the modifications that can be made to some of the molecules of that compound are considered, then, absent a contrary explicit statement, every combination and permutation of the compound and possible modifications are specifically contemplated. Thus, if classes of molecules A, B, and C and classes of molecules D, E, and F are disclosed, and an example of a combination molecule A-D is disclosed, then each is individually and collectively contemplated, meaning that combinations A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are considered to be disclosed, even if each is not individually enumerated. Similarly, any subset or combination of these is also disclosed. Thus, for example, the subgroups A-E, B-F, and C-E are considered to be disclosed. This concept applies to all aspects of this application, including but not limited to the steps in the methods of making and using the disclosed compositions. Thus, if there are various additional steps that are practicable, each of these additional steps is understood to be capable of being carried out by any specific embodiment or combination of embodiments of the disclosed method.

[0015] The various aspects of the present invention are described in further detail in the following sections.

[0016] (I) Compositions This specification provides an assimilable amino acid composition of a composition useful for stimulating muscle protein synthesis. In some embodiments, the composition may include the essential amino acids histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine, and tryptophan. In preferred embodiments, the composition may include the amino acid arginine in addition to the essential amino acids histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine, and tryptophan.

[0017] Generally, the amino acids can be L-amino acids, D-amino acids, or mixtures thereof. In preferred embodiments, the amino acids are L-amino acids. Those skilled in the art will understand that the amino acids of the composition may be free amino acids or amino acid salts. The amino acid composition of the present invention may also be in the form of a complete protein or peptide as long as the protein or peptide contains the amino acids of the present invention in appropriate concentrations relative to each other. In preferred embodiments, the amino acids of the present invention are free amino acids or amino acid salts. If commercially available products are not purchased, the individual amino acids can be produced by methods well known in the art, including chemical synthesis or the use of recombinant microorganisms.

[0018] The amino acids can be the standard amino acids histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine, tryptophan, and arginine, or non-standard amino acid derivatives (e.g., precursors of amino acids). Non-limiting examples of non-standard amino acids that can be used in the present invention include L-lysine acetate, L-tyrosine derivatives, ornithine, keto acid analogs, hydrochloride (L-cysteine HCL.H2O), and N-acetyl derivatives of various amino acids. In preferred embodiments, the amino acids used in the compositions and solutions of the present invention can be in free form or in salt form.

[0019] In one aspect, the amino acid composition of the present disclosure comprises one or more essential amino acids. Essential amino acids (EAAs) are amino acids that the subject cannot synthesize de novo and thus need to be supplemented in the diet. The amino acids considered essential for humans are phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, and histidine. In some embodiments, the combination of EEAs comprises phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, and histidine. In preferred embodiments, the amino acid composition also comprises at least one conditionally essential amino acid.

[0020] Amino acids such as arginine, cysteine, glycine, glutamine, proline, serine, and tyrosine are considered conditionally essential and are not normally considered essential in the diet, but it means that specific groups that do not synthesize these amino acids in sufficient amounts must be supplemented externally. For example, the body produces enough arginine to meet metabolic requirements under normal conditions. Therefore, for many subjects, there is no need to supplement arginine to stimulate muscle protein synthesis. However, in certain clinical situations, including elderly patients with heart failure, endogenous arginine production is insufficient to meet all the needs. For such groups, the combination of the present invention containing arginine or a precursor of the amino acid arginine is advantageous. The term "amino acid precursor" refers to a metabolic precursor of an amino acid. For example, serine is a metabolic precursor of cysteine and glycine, and 3-phosphoglyceric acid is a metabolic precursor of serine and is therefore also a metabolic precursor of cysteine and glycerine. As a second example, citrulline is a metabolic precursor of arginine. Metabolic pathways for synthesizing amino acids are well known in the art, and those skilled in the art can refer to this to identify metabolic precursors of other amino acids. In some embodiments, the combination of the present invention includes phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, histidine, and at least one conditionally essential amino acid. For example, the combination may include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 conditionally essential amino acids. In other embodiments, the combination of the present invention includes phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, histidine, and at least one conditionally essential amino acid precursor. For example, the combination may include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 conditionally essential amino acid precursors. In still other embodiments, the combination of the present invention includes phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, histidine, at least one non-essential amino acid, and at least one conditionally essential amino acid precursor.

[0021] In some embodiments, the composition may contain histidine at the following concentrations, expressed as w / w%: about 1% to about 2%, about 1.1% to about 2%, about 1.2% to about 2%, about 1.3% to about 2%, about 1.4% to about 2%, about 1.5% to about 2%, about 1.6% to about 2%, about 1.7% to about 2%, about 1.8% to about 2%, about 1.9% to about 2%, about 1% to about 1.9%, about 1% to about 1.8%, about 1% to about 1.7%, about 1% to about 1.6%, about 1% to about 1.5%, about 1% to about 1.4%, about 1% to about 1.3%, about 1% to about 1.2%, or about 1% to about 1.1% histidine.

[0022] In some embodiments, the composition may contain isoleucine at the following concentrations, expressed as w / w%: about 9% to about 11%, about 9.1% to about 11%, about 9.2% to about 11%, about 9.3% to about 11%, about 9.4% to about 11%, about 9.4% to about 11%, about 9.4% to about 11%, about 9.5% to about 11%, about 9.6% to about 11%, about 9.7% to about 11%, about 9.8% to about 11%, about 9.9% to about 11%, about 10% to about 11%, about 10.1% to about 11%, about 10.2% to about 11%, about 10.3% to about 11%, about 10.4% to about 11%, about 10.5% to about 11%, about 10.6% to about 11%, about 10.7% to about 11%, about 10.8% to about 11%, about 10.9% to about 11%, about 9% to about 10.9%, about 9% to about 10.8%, about 9% to about 10.7%, about 9% to about 10.6%, about 9% to about 10.5%, about 9% to about 10.4%, about 9% to about 10.3%, about 9% to about 10.2%, about 9% to about 10.1%, about 9% to about 10%, about 9% to about 9.9%, about 9% to about 9.8%, about 9% to about 9.7%, about 9% to about 9.6%, about 9% to about 9.5%, about 9% to about 9.4%, about 9% to about 9.3%, about 9% to about 9.2%, or about 9% to about 9.1% isoleucine.

[0023] In some embodiments, the composition may contain leucine at the following concentrations, expressed as % w / w: from about 35% to about 38%, from about 35.2% to about 38%, from about 35.4% to about 38%, from about 35.6% to about 38%, from about 35.8% to about 38%, from about 36% to about 38%, from about 36.2% to about 38%, from about 36.4% to about 38%, from about 36.6% to about 38%, from about 36.8% to about 38%, from about 37% to about 38%, from about 37.2% to about 38%, from about 37.4% to about 38%, from about 37.6% to about 38%, from about 37.8% to about 38%, from about 35% to about 37.8%, from about 35% to about 37.6%, from about 35% to about 37.4%, from about 35% to about 37.2%, from about 35% to about 37%, from about 35% to about 36.8%, from about 35% to about 36.6%, from about 35% to about 36.4%, from about 35% to about 36.2%, from about 35% to about 36%, from about 35% to about 35.8%, from about 35% to about 35.6%, from about 35% to about 35.4%, or from about 35% to about 35.2% leucine.

[0024] In some embodiments, the composition may contain lysine at the following concentrations, expressed as % w / w: from about 14% to about 17%, from about 14.2% to about 17%, from about 14.4% to about 17%, from about 14.6% to about 17%, from about 14.8% to about 17%, from about 15% to about 17%, from about 15.2% to about 17%, from about 15.4% to about 17%, from about 15.6% to about 17%, from about 15.8% to about 17%, from about 16% to about 17%, from about 16.2% to about 17%, from about 16.4% to about 17%, from about 16.6% to about 17%, from about 16.8% to about 17%, from about 14% to about 16.8%, from about 14% to about 16.6%, from about 14% to about 16.4%, from about 14% to about 16.2%, from about 14% to about 16%, from about 14% to about 15.8%, from about 14% to about 15.6%, from about 14% to about 15.4%, or from about 14% to about 15.2% lysine.

[0025] In some embodiments, the composition may contain methionine at the following concentrations, expressed as w / w%: about 2% to about 4%, about 2.1% to about 4%, about 2.2% to about 4%, about 2.3% to about 4%, about 2.4% to about 4%, about 2.5% to about 4%, about 2.6% to about 4%, about 2.7% to about 4%, about 2.8% to about 4%, about 2.9% to about 4%, about 3% to about 4%, about 3.1% to about 4%, about 3.2% to about 4%, about 3.3% to about 4%, about 3.4% to about 4%, about 3.5% to about 4%, about 3.6% to about 4%, about 3.7% to about 4%, about 3.8% to about 4%, about 3.9% to about 4%, about 2% to about 3.9%, about 2% to about 3.8%, about 2% to about 3.7%, about 2% to about 3.6%, about 2% to about 3.5%, about 2% to about 3.4%, about 2% to about 3.3%, about 2% to about 3.2%, about 2% to about 3.1%, about 2% to about 3%, about 2% to about 2.9%, about 2% to about 2.8%, about 2% to about 2.7%, about 2% to about 2.6%, about 2% to about 2.5%, about 2% to about 2.4%, about 2% to about 2.3%, about 2% to about 2.2%, or about 2% to about 2.1% methionine.

[0026] In some embodiments, the composition may contain phenylalanine at the following concentrations, expressed as w / w%: about 5% to about 7%, about 5.1% to about 7%, about 5.2% to about 7%, about 5.3% to about 7%, about 5.4% to about 7%, about 5.5% to about 7%, about 5.6% to about 7%, about 5.7% to about 7%, about 5.8% to about 7%, about 5.9% to about 7%, about 6% to about 7%, about 6.1% to about 7%, about 6.2% to about 7%, about 6.3% to about 7%, about 6.4% to about 7%, about 6.5% to about 7%, about 6.6% to about 7%, about 6.7% to about 7%, about 6.8% to about 7%, about 6.9% to about 7%, about 5% to about 6.9%, about 5% to about 6.8%, about 5% to about 6.7%, about 5% to about 6.6%, about 5% to about 6.5%, about 5% to about 6.4%, about 5% to about 6.3%, about 5% to about 6.2%, about 5% to about 6.1%, about 5% to about 6%, about 5% to about 5.9%, about 5% to about 5.8%, about 5% to about 5.7%, about 5% to about 5.6%, about 5% to about 5.5%, about 5% to about 5.4%, about 5% to about 5.3%, about 5% to about 5.2%, or about 5% to about 5.1% phenylalanine.

[0027] In some embodiments, the composition may contain threonine at the following concentrations, expressed as w / w%: about 8% to about 9%, about 8.1% to about 9%, about 8.2% to about 9%, about 8.3% to about 9%, about 8.4% to about 9%, about 8.5% to about 9%, about 8.6% to about 9%, about 8.7% to about 9%, about 8.8% to about 9%, about 8.9% to about 9%, about 8% to about 8.9%, about 8% to about 8.8%, about 8% to about 8.7%, about 8% to about 8.6%, about 8% to about 8.5%, about 8% to about 8.4%, about 8% to about 8.3%, about 8% to about 8.2%, or about 8% to about 8.1% threonine.

[0028] In some embodiments, the composition may contain valine at the following concentrations, expressed as w / w%: about 9% to about 11%, about 9.1% to about 11%, about 9.2% to about 11%, about 9.3% to about 11%, about 9.4% to about 11%, about 9.4% to about 11%, about 9.4% to about 11%, about 9.5% to about 11%, about 9.6% to about 11%, about 9.7% to about 11%, about 9.8% to about 11%, about 9.9% to about 11%, about 10% to about 11%, about 10.1% to about 11%, about 10.2% to about 11%, about 10.3% to about 11%, about 10.4% to about 11%, about 10.5% to about 11%, about 10.6% to about 11%, about 10.7% to about 11%, about 10.8% to about 11%, about 10.9% to about 11%, about 9% to about 10.9%, about 9% to about 10.8%, about 9% to about 10.7%, about 9% to about 10.6%, about 9% to about 10.5%, about 9% to about 10.4%, about 9% to about 10.3%, about 9% to about 10.2%, about 9% to about 10.1%, about 9% to about 10%, about 9% to about 9.9%, about 9% to about 9.8%, about 9% to about 9.7%, about 9% to about 9.6%, about 9% to about 9.5%, about 9% to about 9.4%, about 9% to about 9.3%, about 9% to about 9.2%, or about 9% to about 9.1% valine.

[0029] In some embodiments, the composition may contain tryptophan at the following concentrations, expressed as w / w%: from about 0.005% to about 0.8%, from about 0.006% to about 0.8%, from about 0.007% to about 0.8%, from about 0.008% to about 0.8%, from about 0.009% to about 0.8%, from about 0.01% to about 0.8%, from about 0.02% to about 0.8%, from about 0.03% to about 0.8%, from about 0.04% to about 0.8%, from about 0.05% to about 0.8%, from about 0.06% to about 0.8%, from about 0.07% to about 0.8%, from about 0.08% to about 0.8%, from about 0.09% to about 0.8%, from about 0.1% to about 0.8%, from about 0.2% to about 0.8%, from about 0.3% to about 0.8%, from about 0.4% to about 0.8%, from about 0.5% to about 0.8%, from about 0.6% to about 0.8%, from about 0.7% to about 0.8%, from about 0.005% to about 0.7%, from about 0.005% to about 0.6%, from about 0.005% to about 0.5%, from about 0.005% to about 0.4%, from about 0.005% to about 0.3%, from about 0.005% to about 0.2%, from about 0.005% to about 0.1%, from about 0.005% to about 0.09%, from about 0.005% to about 0.08%, from about 0.005% to about 0.07%, from about 0.005% to about 0.06%, from about 0.005% to about 0.05%, from about 0.005% to about 0.04%, from about 0.005% to about 0.03%, from about 0.005% to about 0.02%, from about 0.005% to about 0.01%, from about 0.005% to about 0.009%, from about 0.005% to about 0.008%, from about 0.005% to about 0.007%, or from about 0.005% to about 0.006% of tryptophan.

[0030] In some embodiments, the composition may contain arginine at the following concentrations, expressed as w / w%: about 8% to about 11%, about 8.1% to about 11%, about 8.2% to about 11%, about 8.3% to about 11%, about 8.4% to about 11%, about 8.5% to about 11%, about 8.6% to about 11%, about 8.7% to about 11%, about 8.8% to about 11%, about 8.9% to about 11%, about 9% to about 11%, about 9.1% to about 11%, about 9.2% to about 11%, about 9.3% to about 11%, about 9.4% to about 11%, about 9.4% to about 11%, about 9.4% to about 11%, about 9.5% to about 11%, about 9.6% to about 11%, about 9.7% to about 11%, about 9.8% to about 11%, about 9.9% to about 11%, about 10% to about 11%, about 10.1% to about 11%, about 10.2% to about 11%, about 10.3% to about 11%, about 10.4% to about 11%, about 10.5% to about 11%, about 10.6% to about 11%, about 10.7% to about 11%, about 10.8% to about 11%, about 10.9% to about 11%, about 8% to about 10.8%, about 8% to about 10.7%, about 8% to about 10.6%, about 8% to about 10.5%, about 8% to about 10.4%, about 8% to about 10.3%, about 8% to about 10.2%, about 8% to about 10.1%, about 8% to about 10%, about 8% to about 9.9%, about 8% to about 9.8%, about 8% to about 9.7%, about 8% to about 9.6%, about 8% to about 9.5%, about 8% to about 9.4%, about 8% to about 9.3%, about 8% to about 9.2%, about 8% to about 9.1%, about 8% to about 9%, about 8% to about 8.9%, about 8% to about 8.8%, about 8% to about 8.7%, about 8% to about 8.6%, about 8% to about 8.5%, about 8% to about 8.4%, about 8% to about 8.3%, about 8% to about 8.2%, or about 8% to about 8.1% arginine.

[0031] In some embodiments, the composition may contain amino acids at the following concentrations, expressed as w / w%: about 1% to about 2% histidine, about 9% to about 11% isoleucine, about 35% to about 38% leucine, about 14% to about 17% lysine, about 2% to about 4% methionine, about 5% to about 7% phenylalanine, about 8% to about 9% threonine, about 9% to about 11% valine, about 0.005% to about 0.8% tryptophan, and about 8% to about 11% arginine. In an exemplary embodiment, the composition may comprise, consist essentially of, or consist of 1.5% histidine, 9.7% isoleucine, 36.4% leucine, 15.2% lysine, 3% methionine, 6.1% phenylalanine, 8.5% threonine, 10% valine, 0.6% tryptophan, and 9% arginine.

[0032] In another aspect, the combination of the present invention may further include one or more additional nutrients. As used herein, the term "nutrient" refers to prebiotics, vitamins, carbohydrates, fiber, fatty acids, sulfates, minerals, antioxidants, and other food components that are taken up from the internal environment and used in the metabolism of the subject. Suitable vitamins can include, but are not limited to, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B9, vitamin B12, lipoic acid, vitamin A, biotin, vitamin K, vitamin C, vitamin D, and vitamin E. Suitable minerals can include, but are not limited to, compounds containing iron, copper, magnesium, manganese, molybdenum, nickel, and zinc. Suitable enzyme cofactors can include, but are not limited to, adenosine triphosphate (ATP), S-adenosylmethionine (SAM), coenzyme B, coenzyme M, coenzyme Q, glutathione, heme, methanofuran, and nucleotide sugars. Suitable lipids can include, but are not limited to, fatty acids, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids, prenol lipids, saccharolipids, and polyketides. Non-limiting examples of fatty acids include myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, hexadecatrienoic acid, alpha-linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, heneicosapentaenoic acid, docosapentaenoic acid, curpanodonic acid, docasehaenoic acid, tetracosapentaenoic acid, and tetracosahexaenoic acid.Non-limiting examples of nutrient addition may include thiamine, riboflavin, niacin, folic acid, pantothenic acid, calcium, phosphorus, magnesium, manganese, iron, zinc, copper, selenium, sodium, potassium, beta-carotene, retinol, alpha-tocopherol, beta-tocopherol, gamma-tocopherol, delta-tocopherol, alpha-tocotrienol, beta-tocotrienol, gamma-tocotrienol, delta-tocotrienol, apo-8-carotenal, trans-lycopene, cis-lycopene, trans-beta-carotene, and cis-beta-carotene, and caffeine. In some embodiments, the combination of the present invention further comprises one nutrient. In other embodiments, the combination of the present invention further comprises at least one nutrient. For example, the combination of the present invention may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more nutrients. In still other embodiments, the combination of the present invention further comprises two or more nutrients. In a preferred embodiment, the combination of the present invention further comprises at least one nutrient selected from the group consisting of omega-3 fatty acids and biotin. In other preferred embodiments, the combination of the present invention further comprises two nutrients selected from the group consisting of omega-3 fatty acids and biotin. The total contribution of one or more nutrients to the total weight of the combination is substantially less than the contribution of a plurality of amino acids. Generally, one or more nutrients constitute about 10% by weight or less, preferably about 5% by weight or less, more preferably about 3% by weight or less of the combination. In a preferred embodiment, one or more nutrients are biotin. In other preferred embodiments, one or more nutrients are one or more omega-3 fatty acids. In other preferred embodiments, one or more nutrients are biotin and one or more omega-3 fatty acids.

[0033] (II) Formulation In each of the above embodiments, the amino acids and nutrients (if present) can be formulated for animal or human use. In some embodiments, each amino acid and nutrient (if present) is formulated separately. In other embodiments, two or more amino acids and nutrients (if present) are formulated together. In still other embodiments, all of the amino acids and nutrients that make up the combination of the present invention are formulated together. One or more formulations can be further processed into one or more dosage forms and administered simultaneously, sequentially, or over a period of time (e.g., over 1 minute, 10 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 9 hours, 12 hours, 18 hours, 24 hours, or more). Administration can be carried out using effective standard techniques including oral, parenteral (e.g., intravenous, intraperitoneal, subcutaneous, intramuscular), buccal, sublingual, or suppository administration. As used herein, the term oral refers to any form of oral administration, including addition of the composition to animal feed or other food. For the formulation of pharmaceutical compositions, see, for example, Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. (1975), and Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y. (1980).

[0034] Methods for preparing compositions for animal or human use are well known in the art. For example, compositions can generally be formulated as liquid compositions, solid compositions, or semi-solid compositions. Liquid compositions include, but are not limited to, suspension aqueous solutions, solutions, emulsions, elixirs, or syrups. Liquid compositions are typically considered to include a solvent carrier selected from polar solvents, non-polar solvents, or a combination of both. The choice of solvent is influenced by the properties of the components of the composition. For example, if the components are water-soluble, a polar solvent may be used. Alternatively, if the components of the composition are fat-soluble, a non-polar solvent may be used. Suitable polar and non-polar solvents are known in the art. Semi-solid compositions include irrigation solutions, suppositories, creams, and topical agents. Dry compositions include, but are not limited to, reconstitutable powders, chewable tablets, instant tablets, effervescent tablets, multi-layer tablets, bilayer tablets, capsules, soft gelatin capsules, hard gelatin capsules, caplets, troches, chewable troches, beads, powders, granules, pellets, microparticles, and dispersible granules. The formulations can include the combinations of the present invention together with excipients. Non-limiting examples of excipients include binders, diluents (bulking agents), disintegrants, effervescent disintegrants, preservatives (antioxidants), flavor modifiers, lubricants and glidants, dispersants, colorants, pH adjusters, chelating agents, antibacterial agents, release control polymers, and combinations of any of these agents.

[0035] Non-limiting examples of binders suitable for the formulations of various embodiments include starch, pregelatinized starch, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamide, polyvinyl oxazolidone, polyvinyl alcohol, C12-C18 fatty alcohols, polyethylene glycol, polyols, saccharides, oligosaccharides, polypeptides, oligopeptides, and combinations thereof. The polypeptide can be amino acids in any sequence in the range of about 100 to about 300,000 daltons. In one embodiment, the binder can be introduced into the mixture and granulated into a solid form, which includes, but is not limited to, crystals, particles, powders, or any other micronized solid form known in the art. In another embodiment, the binder can be dissolved or suspended in a solvent and sprayed as a liquid binder into the mixture in the granulator during granulation.

[0036] Non-limiting examples of diluents (also referred to as "filler" or "diluent") include carbohydrates, inorganic compounds, and biocompatible polymers such as polyvinylpyrrolidone (PVP). Other non-limiting examples of diluents include dibasic calcium sulfate, tribasic calcium sulfate, starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, dicalcium phosphate, tricalcium phosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc, modified starch, saccharides such as sucrose, dextrose, lactose, microcrystalline cellulose, fructose, xylitol, and sorbitol, polyhydric alcohols; starch; pre-manufactured direct compression diluents; and mixtures of any of the foregoing.

[0037] The disintegrant may be effervescent or non-effervescent. Non-limiting examples of non-effervescent disintegrants include starches such as corn starch, potato starch, pregelatinized starch, and their modified starches, sweeteners, clays such as bentonite, microcrystalline cellulose, alginates, sodium starch glycolate, gums such as agar, guar, locust bean, karaya, pectin, and tragacanth. Suitable effervescent disintegrants include, but are not limited to, sodium bicarbonate combined with citric acid and sodium bicarbonate combined with tartaric acid.

[0038] Non-limiting examples of preservatives include ascorbic acid and its salts, ascorbyl palmitate, ascorbyl stearate, anoxomer, N-acetylcysteine, benzyl isothiocyanate, m-aminobenzoic acid, o-aminobenzoic acid, p-aminobenzoic acid (PABA), butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), caffeic acid, canthaxanthin, alpha-carotene, beta-carotene, beta-cryptoxanthin, beta-apo-carotenic acid, carnosol, carvacrol, catechin, cetyl gallate, chlorogenic acid, citric acid and its salts, clove extract, coffee bean extract, p-coumaric acid, 3,4-dihydroxybenzoic acid, N,N'-diphenyl-p-phenylenediamine (DPPD), dilauryl thiodipropionate, distearyl thiodipropionate, 2,6-di-tert-butylphenol, dodecyl gallate, edetic acid, ellagic acid, erythorbic acid, sodium erythorbate, esculetin, esculin, 6-ethoxy-1,2-dihydro-2,2,4-trimethylquinoline, ethyl gallate, ethyl maltol, ethylenediaminetetraacetic acid (EDTA), eucalyptus extract, eugenol, ferulic acid, flavonoids (e.g., catechin, epicatechin, epicatechin gallate, epigallocatechin (EGC), epigallocatechin gallate (EGCG), polyphenol epigallocatechin-3-gallate), flavones (e.g., apigenin, chrysin, luteolin), flavonols (e.g., datiscetin, myricetin, daemfero), flavanone, quercetin, fumaric acid, gallic acid, gentiana extract, gluconic acid, glycine, guaiac gum, hesperetin, alpha-hydroxybenzylphosphinic acid, hydroxycinnamic acid, hydroxyglutaric acid, hydroquinone, N-hydroxy succinic acid, hydroxytyrosol, hydroxyurea, rice bran extract, lactic acid and its salts, lecithin, lecithin citrate; R-alpha-lipoic acid, lutein, lycopene, malic acid, maltol, 5-methoxytryptamine, methyl gallate, citric acid monoglyceride; citric acid monoisopropyl;Morin, beta-naphthoflavone, nordihydroguaiaretic acid (NDGA), octyl gallate, oxalic acid, palmitoyl citrate, phenothiazine, phosphatidylcholine, phosphoric acid, phosphate, phytic acid, phytyl bichromel, pimento extract, propyl gallate, polyphosphate, quercetin, trans-resveratrol, rosemary extract, rosmaric acid, sage extract, sesamol, silymarin, sinapic acid, succinic acid, stearyl citrate, syringic acid, tartaric acid, thymol, tocopherol (i.e., alpha-tocopherol, beta-tocopherol, gamma-tocopherol, and delta-tocopherol), tocotrienol (i.e., alpha-tocotrienol, beta-tocotrienol, gamma-tocotrienol, and delta-tocotrienol), tyrosol, vanillic acid, 2,6-di-tert-butyl-4-hydroxymethylphenol (i.e., Ionox 100), 2,4-(tris-3´,5´-bi-tert-butyl-4´-hydroxybenzyl)-mesitylene (i.e., Ionox 330), 2,4,5-trihydroxybutyrophenone, ubiquinone, tert-butylhydroquinone (TBHQ), thiodipropionic acid, trihydroxybutyrophenone, tryptamine, tyramine, uric acid, vitamin K and derivatives, vitamin Q10, wheat germ oil, zeaxanthin, or combinations thereof, but not limited thereto. In an exemplary embodiment, the preservative is an antioxidant such as a-tocopherol or ascorbic acid, and an antibacterial agent such as parabens, chlorobutanol, or phenol.;

[0039] Suitable flavor modifiers include flavoring agents, taste-correcting agents, sweeteners, and the like. Examples of flavoring agents include, but are not limited to, synthetic essential oils and flavoring fragrances and / or extracts from natural oils, plants, leaves, flowers, fruits, and combinations thereof. Other non-limiting examples of flavorings include cinnamon oil, wintergreen oil, peppermint oil, clover oil, hay oil, anise oil, eucalyptus, vanilla, lemon oil, orange oil, citrus oils such as grape oil and grapefruit oil, and fruit essential oils such as apple, peach, pear, strawberry, raspberry, cherry, plum, pineapple, and apricot.

[0040] As flavoring agents, hydroxypropyl ethers (HPC) of cellulose, such as Klucel (registered trademark), Nisswo HPC, and PrimaFlo HP22; low-substitution hydroxypropyl ethers (L-HPC); hydroxypropyl methyl ethers (HPMC) of cellulose, such as Seppifilm-LC, Pharmacoat (registered trademark), Metolose SR, Opadry YS, PrimaFlo, MP3295A, Benecel MP824, and Benecel MP843; methylcellulose polymers, such as Methocel (registered trademark) and Metolose (registered trademark); ethylcellulose (EC) and its mixtures, such as E461, Ethocel (registered trademark), Aqualon (registered trademark)-EC, Surelease; polyvinyl alcohol (PVA) such as Opadry AMB; hydroxyethylcellulose such as Natrosol (registered trademark); carboxymethylcellulose and salts of carboxymethylcellulose (CMC), such as Aualon (registered trademark)-CMC; polyvinyl alcohol and polyethylene glycol copolymers, such as Kollicoat IR (registered trademark); monoglycerides (Myverol), triglycerides (KLX), polyethylene glycol, modified food starch, acrylic polymers, and mixtures of acrylic polymers and cellulose ethers, such as Eudragit (registered trademark) EPO, Eudragit (registered trademark) RD100, and Eudragit (registered trademark) E100; cellulose acetate phthalate; Sepifilm, cyclodextrin, such as a mixture of HPMC and stearic acid, and mixtures of these materials are included, but not limited to these. In other embodiments, additional flavoring agents contemplated are those described in U.S. Patent Nos. 4,851,226, 5,075,114, and 5,876,759, each of which is hereby incorporated by reference in its entirety.

[0041] Non-limiting examples of sweeteners include glucose (corn syrup), dextrose, invert sugar, fructose, and mixtures thereof (when not used as a carrier); saccharin and its various salts, such as the sodium salt; dipeptide sweeteners such as aspartame; dihydrochalcone compounds, glycyrrhizin; Stevia rebaudiana (stevioside); chloro derivatives of sucrose, such as sucralose; sugar alcohols, such as sorbitol, mannitol, xylitol, hydrolyzed hydrogenated starch, and the synthetic sweetener 3,6-dihydro-6-methyl-1,2,3-oxathiazin-4-one-2,2-dioxide, particularly the potassium salt (acesulfame K), as well as its sodium and calcium salts.

[0042] The components can be lubricated using a lubricant to form the composition of the present invention. The lubricant as a lubricant facilitates the removal of the solid dosage form during the manufacturing process. Non-limiting examples of lubricants and lubricating agents include magnesium stearate, calcium stearate, zinc stearate, hydrogenated vegetable oil, sterotex, polyoxyethylene monostearate, talc, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, magnesium lauryl sulfate, and light mineral oil. The composition generally contains from about 0.01% to about 10% by weight of the lubricant. In some embodiments, the composition contains from about 0.1% to about 5% by weight of the lubricant. In a further embodiment, the composition contains from about 0.5% to about 2% by weight of the lubricant.

[0043] The dispersant may include, but is not limited to, starch, alginic acid, polyvinylpyrrolidone, guar gum, kaolin, bentonite, purified wood cellulose, sodium starch glycolate, isomorphous silicate, and microcrystalline cellulose, which is an emulsifying surfactant with a high hydrophilic-lipophilic balance (HLB).

[0044] In some embodiments, it may be desirable to include a colorant. Suitable color additives include, but are not limited to, Food, Drug, and Cosmetic Colors (FD&C), Drug and Cosmetic Colors (D&C), or External Drug and Cosmetic Colors (Ext. D&C). These colors or dyes and their corresponding lakes, as well as certain natural and derived colorants, may be suitable for use in various embodiments.

[0045] Non-limiting examples of pH adjusters include citric acid, acetic acid, tartaric acid, malic acid, fumaric acid, lactic acid, phosphoric acid, sorbic acid, benzoic acid, sodium carbonate, and sodium bicarbonate.

[0046] To inhibit oxidative degradation of the morphinan by the oxidative groups, chelating agents may be included as excipients to immobilize the oxidative groups, including, but not limited to, metal ions. Non-limiting examples of chelating agents include lysine, methionine, glycine, gluconate, polysaccharides, glutamate, aspartate, and disodium ethylenediaminetetraacetate (Na 2 EDTA).

[0047] Antimicrobial agents can be included as excipients to minimize degradation of the compounds described herein by microbial agents, including, but not limited to, bacteria and fungi. Non-limiting examples of antimicrobial agents include parabens, chlorobutanol, phenol, calcium propionate, sodium nitrate, sodium nitrite, Na 2 These include EDTA, and sulfites, including but not limited to sulfur dioxide, sodium bisulfite, and potassium bisulfite.

[0048] In various embodiments of the solid dosage form compositions incorporating the compounds according to the present disclosure, a release control polymer may be included. In one embodiment, the release control polymer can be used as a tablet coating. In other embodiments including, but not limited to, two-layer tablets, the release control polymer may be mixed with granules and other excipients prior to tablet formation by known processes including, but not limited to, compression into tablet form. Suitable release control polymers include, but are not limited to, hydrophilic polymers and hydrophobic polymers.

[0049] Suitable hydrophilic release control polymers include, but are not limited to, cellulose acetate, cellulose diacetate, cellulose triacetate, cellulose ethers, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, nitrocellulose, crosslinked starch, agar, casein, chitin, collagen, gelatin, maltose, mannitol, maltodextrin, pectin, pullulan, sorbitol, xylitol, polysaccharides, ammonium alginate, sodium alginate, calcium alginate, potassium alginate, propylene glycol alginate, sodium carboxymethylcellulose alginate, calcium carboxymethylcellulose, carrageenan, fucoidan, gellan gum, gum arabic, carrageenan gum, gatti gum, guar gum, karaya gum, locust bean gum, okra gum, tragacanth gum, scleroglucan gum, xanthan gum, laminariaceae, laminaran, acrylic polymers, acrylate polymers, carboxyvinyl polymers, copolymers of maleic anhydride and styrene, copolymers of maleic anhydride and ethylene, copolymers of maleic anhydride and propylene or copolymers of maleic anhydride and isobutylene, crosslinked polyvinyl alcohol and poly N-vinyl-2-pyrrolidone, diesters of polyglucan, polyacrylamides, polyacrylic acid, polyamides, polyethylene glycol, polyethylene oxides, poly(hydroxyalkyl methacrylate), polyvinyl acetate, polyvinyl alcohol, polyvinyl chloride, polystyrene, polyvinyl pyrrolidone, anionic and cationic hydrogels, and combinations thereof.

[0050] The amino acid compositions disclosed herein may also include compositions that can be made as powders that can be added to foods such as baked products (e.g., cookies and brownies), and as concentrates. The concentrate can be added to water or other ingestible liquids to make a nutritional beverage. Nutritional supplements are typically contained in single - serving or multi - serving containers such as packages, boxes, cartons, wrappers, bottles, or cans. When preparing a nutritional supplement in the form of a concentrate that can be added to and mixed with a beverage, bottles or cans can be used for packaging the concentrate. A nutritional supplement may contain water.

[0051] (a) Amino acid dosage As will be appreciated by those skilled in the art, the dosage of the amino acid compositions of the present invention can and will vary depending on the weight, sex, age, and / or medical condition of the subject, the intensity of physical activity by the subject, and the method of administration. Non - limiting examples of species can include humans, companion animals, laboratory animals, zoo animals, or agricultural animals. The required dosage can be readily determined by routine experimentation. A typical amino acid dosage for oral administration can be about 7 g per dose. In some embodiments, an amino acid composition with an amino acid dosage of about 3, 4, 5, 10, or 12 g can be administered. In other embodiments, an amino acid composition with an amino acid dosage of about 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 g can be administered. In an exemplary embodiment, the amino acid dosage can be about 7 g per dose.

[0052] Also, to provide the desired stimulation of muscle protein synthesis, multiple doses of the amino acid composition per day can be used as needed. For example, one, two, three, four, or more doses of the amino acid composition can be administered per day. In a preferred embodiment, one dose of the amino acid composition can be administered per day. In another preferred embodiment, two doses of the amino acid composition can be administered per day. In yet another preferred embodiment, three doses of the amino acid composition can be administered per day.

[0053] The timing and duration of administration of the composition of the present invention can be changed and will be different. For example, when administering the composition to stimulate muscle protein synthesis regardless of the presence or absence of exercise, the composition can be administered without regular exercise, before starting regular exercise, during regular exercise, or after regular exercise. Alternatively, when administering the composition to improve muscle protein synthesis in a subject prone to muscle problems, such as an elderly human subject, the composition can be administered regularly.

[0054] In an exemplary embodiment, when the subject is human, 3 g of the amino acid composition per dose can be administered three times a day. In another exemplary embodiment, 7 g of the amino acid composition per dose can be administered twice a day. In yet another exemplary embodiment, 7 g of the amino acid composition per dose can be administered once a day.

[0055] (b) Administration The composition of the present invention can be administered by intravenous injection, intramuscular injection, subcutaneous injection, or parenterally. In some embodiments, the composition can be formulated for administration by injection (e.g., intraperitoneal, intravenous, subcutaneous, intramuscular, etc.). Accordingly, these compositions are preferably combined with a pharmaceutically acceptable vehicle such as physiological saline, Ringer's solution, dextrose solution, etc.

[0056] The amino acid composition of the present invention may contain a pharmaceutical carrier (or excipient). Such a carrier may be any solvent or a solid material for encapsulation, and is non-toxic. The carrier may be something that forms a shape or imparts viscosity, or may act as a diluent. Suitable pharmaceutical carriers may include liquid carriers such as normal saline and other non-toxic salts at physiological concentrations or approximations thereof, as well as solid carriers not used in humans such as talc or sucrose, or animal feed. The carrier may also include stabilizers, wetting agents, and emulsifiers, salts for osmotic pressure changes, encapsulating agents, buffering agents, and skin penetration enhancers. Carriers and excipients, as well as formulations for parenteral and oral drug delivery, are described in Remington’s Pharmaceutical Sciences 19th Ed. Mack Publishing (1995).

[0057] For parenteral administration (including subcutaneous, intradermal, intravenous, intramuscular, and intraperitoneal), the composition may be an aqueous solution or an oily solution. The aqueous solution may include water, a sterile diluent such as physiological saline, a pharmaceutically acceptable polyol such as glycerol, propylene glycol, or other synthetic solvents; an antibacterial and / or antifungal agent such as benzyl alcohol, methylparaben, chlorobutanol, phenol, thimerosal; an antioxidant such as ascorbic acid or sodium bisulfite; a chelating agent such as ethylenediaminetetraacetic acid; a buffer such as acetate, citrate, or phosphate; and / or an agent for isotonicity adjustment such as sodium chloride, dextrose, or a polyhydric alcohol such as mannitol or sorbitol. The pH of the aqueous solution can be adjusted with an acid or a base such as hydrochloric acid or sodium hydroxide. The oily solution or suspension may further include sesame oil, peanut oil, olive oil, or mineral oil.

[0058] In a preferred embodiment, the amino acid composition of the present invention can be administered orally. Non-limiting examples of oral formulations that can be used for the administration of the amino acid composition of the present invention include nutritional formulations, medical foods, medical beverages, complete food forms, partial food forms, food additives in the form of powders suitable for decomposition, pharmaceutical formulations in the form of tablets, pills, sachets, or capsules, or those by means of enteral nutrition, such as nasogastric tubes, nasoduodenal tubes, esophageal fistula tubes, gastric fistula tubes, or jejunal fistula tubes, or by means of peripheral or total parenteral nutrition methods. In an exemplary embodiment, the composition of the present invention can be administered orally as a dietary supplement.

[0059] Compositions for oral administration generally contain an inert excipient in addition to the amino acid component of the composition. Oral preparations can also be enclosed in gelatin capsules or compressed into tablets. Common excipients used in such preparations include pharmaceutically compatible bulking agents / diluents, such as microcrystalline cellulose, hydroxypropylmethylcellulose, starch, lactose, sucrose, glucose, mannitol, sorbitol, dibasic calcium phosphate, or calcium carbonate; binders, such as alginic acid, carboxymethylcellulose, microcrystalline cellulose, gelatin, tragacanth gum, or polyvinylpyrrolidone; disintegrants, such as alginic acid, cellulose, starch, or polyvinylpyrrolidone; lubricants, such as calcium stearate, magnesium stearate, talc, silica, or sodium stearyl fumarate; lubricants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; flavoring agents, such as peppermint, methyl salicylate, or citrus flavors; coloring agents; and preservatives, such as antioxidants (e.g., vitamin A, vitamin C, vitamin E, or retinyl palmitate), citric acid, or sodium citrate. Oral preparations can also be administered as aqueous suspensions, elixirs, or syrups. In that case, various sweeteners or flavoring agents, coloring agents, and, if desired, emulsifying agents and / or suspending agents, and diluents such as water, ethanol, glycerin, and combinations thereof can be combined with the active ingredient.

[0060] The composition according to the invention can be nutritionally complete. That is, in addition to vitamins, minerals, trace elements, it can contain a source of nitrogen, carbohydrates, and fats and / or fatty acids so as to be used as a single nutritional source that supplies substantially all of the essential daily amounts of vitamins, minerals, carbohydrates, fats and / or fatty acids, proteins, etc.

[0061] (II) Method The invention also provides a method for increasing muscle protein synthesis, a method for preventing and / or treating a decrease in any one of muscle mass, muscle strength, muscle function, and physical function, or any combination thereof. The method includes administering to a subject a composition comprising a plurality of essential amino acids and optionally non-essential amino acids and / or nutrients. Suitable combinations and formulations for administration are described above in items I and II respectively.

[0062] The nutritional composition according to the present disclosure can advantageously be used in the manufacture of a medicament for the prevention or treatment of a disease or condition associated with muscle loss in a mammal, particularly an adult mammal. Alternatively, the nutritional composition according to the invention can advantageously be used in the manufacture of a medicament for the prevention or treatment of a disease or condition selected from sarcopenia, muscle loss, insufficient muscle protein synthesis, muscle breakdown, muscle proteolysis, muscle atrophy, muscular dystrophy, muscle catabolism, muscle wasting, decrease in muscle strength, decrease in muscle mass, decrease in muscle function, decrease in physical ability, decrease in physical performance, movement disorder, frailty, surgery, physical disability, risk of falling, and risk of fracture associated with falling. Suitable subjects can include humans, domestic animals, companion animals, laboratory animals, or zoological animals. In a preferred embodiment, the subject is a human. The degree of response may vary, in part, depending on a given dose, the exact amino acid combination, the physiological state and / or age of the subject, and / or the timing of administration in relation to the performance of exercise.

[0063] Preferably, the subject is an elderly human. In this regard, in the context of the present application, an elderly human is a person aged 50 years or older, particularly 55 years or older, more particularly 60 years or older, and even more particularly 65 years or older. This somewhat broad definition takes into account the fact that the average age varies between different populations, different continents, etc. Most of the advanced countries in the world consider a calendar age of 65 years as the definition of "elderly" or old people (related to the start of eligibility for pension benefits), but like many Western concepts, it does not fit well with the situation in Africa, for example. At present, there is no standard numerical criterion of the United Nations (UN), but in the UN consensus, when referring to the elderly population in the Western world, 60 years + is used as the lower limit. The traditional definition of the elderly or "aged" people in Africa corresponds to a calendar age of 50 - 65 years, depending on the living environment, region, and country.

[0064] The nutritional composition according to the present disclosure can advantageously be used to prevent or treat muscle loss, particularly loss of muscle mass, during or after a weight maintenance period, during or after a calorie restriction period, during or after bed rest, or during the recovery period after physical trauma. In a particularly preferred embodiment, the composition of the present invention is used for the treatment of a subject, such as a subject suffering from overweight or obesity, after a weight loss program, a calorie restriction program, and / or an exercise program. The subject can be a pediatric, adolescent, adult, or elderly subject. In one embodiment, the subject can be a pediatric, adolescent, or adult.

[0065] Muscle mass and muscle function gradually decline with aging, and as a result, many of the originally healthy human subjects reach a critical point where the function begins to be affected by the age of 60 years. In one aspect, by administering the composition of the present invention to any healthy subject, an increase in muscle protein synthesis, an increase in muscle strength, an increase in muscle function, or a combination thereof is possible, but the degree of response may vary depending on the age and / or physiological state of the subject.

[0066] A decrease in body protein often accompanies a disease, regardless of whether the disease is short-term or long-term. In another aspect, by administering the combination of the present invention to a subject whose body protein has decreased as a result of a disease, an increase in muscle protein synthesis, an increase in muscle strength, an increase in muscle function, or a combination thereof in the subject is possible. In another aspect, by administering the combination of the present invention to a subject at risk of or suffering from muscle atrophy, an increase in muscle protein synthesis, an increase in muscle strength, an increase in muscle function, or a combination thereof is possible. Non-limiting examples of subjects at risk of or suffering from muscle atrophy include humans with a sedentary lifestyle, sedentary workers, medical conditions with restricted movement, bedridden subjects, subjects away from the Earth's gravitational field, subjects with damage to the nerves connecting to the muscles, and subjects at risk of or diagnosed with a disease affecting the nerves controlling the muscles.Non-limiting examples of diseases, disorders, or conditions that cause muscle atrophy include polio, Guillain-Barré syndrome, chronic obstructive pulmonary disease, congestive heart failure, acute coronary syndrome, chronic heart failure, cardiac cachexia, cancer, sarcopenia, spinal cord injury, osteoarthritis, arthritis, stroke, malnutrition, muscular dystrophy (e.g., Becker type, congenital, Duchenne type, distal type, Emery-Dreifuss type, facioscapulohumeral type, limb-girdle type, oculopharyngeal type), spinal muscular atrophy (e.g., ALS, motor neuron disease, infantile progressive spinal muscular atrophy, intermediate spinal muscular atrophy, juvenile spinal muscular atrophy, adult spinal muscular atrophy), inflammatory muscle diseases (e.g., dermatomyositis, polymyositis), peripheral nerve diseases (e.g., Charcot-Marie-Tooth disease, Dejerine-Sottas disease, Friedreich's ataxia), neuromuscular junction diseases (myasthenia gravis, Lambert-Eaton syndrome), muscle metabolic diseases (acid maltase deficiency, carnitine deficiency, carnitine palmitoyltransferase deficiency, debrancher enzyme deficiency, lactate dehydrogenase deficiency, mitochondrial deaminase deficiency, phosphorylase deficiency, phosphofructokinase deficiency, phosphoglycerate kinase deficiency), other muscle diseases (central core myopathy, hyperthyroid myopathy, congenital myotonia, myotubular myopathy, congenital paramyotonia, hypokalemic-hyperkalemic periodic paralysis), or any other disease or condition in which the subject needs to be immobilized or bedridden, particularly chronic heart failure and cardiac cachexia.

[0067] The methods of the invention generally comprise administering an amino acid composition to a subject. Suitable subjects include animals such as mammals and humans. Non-limiting examples of suitable animals include companion animals such as cats, dogs, rodents, and horses; laboratory animals such as mice, rats, and other rodents; agricultural animals such as dairy cows, cattle, pigs, goats, sheep, horses, deer, chickens, and other poultry; zoo animals; and primates such as chimpanzees, monkeys, and gorillas. In a preferred embodiment, the subject is a human. In another preferred embodiment, the subject is a mammal.

[0068] The dosing frequency can be once, twice, three times, or more per day, once, twice, three times, or more per week or month, as necessary to achieve the desired effect. The dosage administered to the subject can vary and will differ depending on the subject (e.g., age, weight, health status) and the particular mode of administration. Those skilled in the art will also understand that the dosage can be determined in accordance with the guidance of Goodman & Goldman’s The Pharmacological Basis of Therapeutics, Tenth Edition (2001), Appendix II, pp. 475-493, and the Physicians’ Desk Reference.

[0069] The human subject can be of any age. In some embodiments, the human subject can be about 30, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or older. In some preferred embodiments, the human subject is 40 years or older. In other preferred embodiments, the human subject is 45 years or older. In still other preferred embodiments, the human subject is 50 years or older. In yet other preferred embodiments, the human subject is 55 years or older. In other preferred embodiments, the human subject is 60 years or older. In still other preferred embodiments, the human subject is 65 years or older. In yet other preferred embodiments, the human subject is 70 years or older. In other preferred embodiments, the human subject is 75 years or older. In still other preferred embodiments, the human subject is 80 years or older. In yet other preferred embodiments, the human subject is 85 years or older. In still other preferred embodiments, the human subject is 90 years or older.

[0070] In some embodiments, the subject can be an option for embodiments for subjects with insufficient daily exercise.

[0071] In other embodiments, the subject can be an option for embodiments for subjects lacking the ability to efficiently stimulate muscle protein synthesis.

[0072] In other embodiments, the subject may be recovering from a lack of physical activity. During high-intensity muscle exercise, the speed of the muscle exercise performed may exceed the ability of the blood supply to deliver oxygen, which can induce reduction stress in muscle cells and lead to conditions that can result in muscle damage, impairment of physical ability, predisposition to injury, and prolonged recovery periods. Non-limiting examples of physical activity that can result in muscle damage, impairment of physical ability, predisposition to injury, and prolonged recovery periods include aerobic exercises such as stretching, cycling, swimming, walking, skipping, rowing, running, hiking, or tennis, weight training, functional training, eccentric training or sprinting, strength training, agility training, or anaerobic exercises such as eccentric training.

[0073] (a) Improvement in muscle protein synthesis The method of the present invention improves muscle protein synthesis in a subject given an amino acid composition as compared to a subject not given the amino acid composition.

[0074] The increase in muscle protein synthesis can be measured by any method known in the art. For example, the rates of protein synthesis and breakdown in muscle and blood have been measured over the years using radioactively labeled amino acids or amino acids labeled with stable isotopes. See, for example, Yale J Biol Med 1997;70(1):65-76, which is incorporated herein by reference. Measurement of 3-methylhistidine excreted in urine or the difference in 3-methylhistidine between arterial and venous blood has been used as a measure of the unidirectional rate of muscle protein breakdown. Total body skeletal mass can be measured using dual energy X-ray (DEXA) absorptiometry or by CT. See, for example, Am J Clin Nutr 2002;76:378-83, and J Appl Physiol 1985, which are incorporated herein by reference. Methods for measuring muscle strength or muscle function are also well known in the art. For example, changes in muscle strength and muscle function can be evaluated using the basic standard tests described in B J Nutr 2012;108:S88-93 or Lu et al. (2012) “Strength and Functional Measurement for Patients with Muscular Dystrophy” Muscular Dystrophy, Dr.Madhuri Hegde (Ed.), each of which is incorporated herein by reference.

[0075] In some embodiments, the method of the present invention can increase muscle protein synthesis by at least two-fold. For example, muscle protein synthesis can increase by at least two-fold, at least three-fold, at least four-fold, at least five-fold, at least six-fold, at least seven-fold, at least eight-fold, at least nine-fold, at least ten-fold, or more after administration of the combination of the present invention. In some embodiments, the method of the present invention can increase muscle strength by at least two-fold. For example, muscle strength can increase by at least two-fold, at least three-fold, at least four-fold, at least five-fold, at least six-fold, at least seven-fold, at least eight-fold, at least nine-fold, at least ten-fold, or more after administration of the combination of the present invention. In some embodiments, the method of the present invention can increase muscle function by at least two-fold. For example, the synthesis of muscle function can increase by at least two-fold, at least three-fold, at least four-fold, at least five-fold, at least six-fold, at least seven-fold, at least eight-fold, at least nine-fold, at least ten-fold, or more after administration of the combination of the present invention. In other embodiments, the method of the present invention can increase muscle protein synthesis, muscle strength, and muscle function, or combinations thereof. Methods for measuring muscle protein synthesis, muscle strength, and muscle function are known in the art and will be described in further detail in the examples.

[0076] (III) Kit Also provided is a kit. Such a kit may include the composition described herein and, in certain embodiments, may include instructions for administration. Such a kit can facilitate the practice of the methods described herein. When supplied as a kit, the various components of the composition can be packaged in separate containers and mixed immediately prior to use. As described herein, the components include, but are not limited to, an amino acid composition. Individual packaging of such components can be provided in the form of a pack or dispenser device that can accommodate one or more unit dosage forms containing the composition, as needed. The pack can include, for example, a metal or plastic foil such as a blister pack. Individual packaging of such components can also, in certain cases, allow for long-term storage without impairing the activity of the components.

[0077] The kit may also include reagents, such as sterile water or sterile saline solution, which are added to, for example, individually packaged lyophilized active ingredients, in separate containers. For example, the lyophilized component can be contained in a sealed glass ampoule, and another ampoule can contain sterile water, sterile saline solution, or a sterile substance, each packaged under a neutral non-reactive gas such as nitrogen. The ampoule can be made of any suitable material, such as glass, an organic polymer (such as polycarbonate, polystyrene, etc.), ceramic, metal, or any other material commonly used for holding reagents. Other examples of suitable containers include bottles that can be manufactured from materials similar to those of ampoules, and enclosures that can consist of the interior lined with foil such as aluminum or an alloy. Other containers include test tubes, vials, flasks, bottles, syringes, etc. The container may have a sterile access port, such as a bottle with a stopper that can be penetrated by a hypodermic needle. Other containers may have two compartments separated by an easily removable membrane that allows the components to be mixed upon removal. The removable membrane can be made of glass, plastic, rubber, etc.

[0078] In certain embodiments, the kit can be supplied with instructions for use. The instructions can be printed on paper or other substrates and / or supplied as an electronically readable medium such as a floppy disk, mini CD-ROM, CD-ROM, DVD-ROM, Zip disk, video tape, audio tape, etc. The detailed instructions need not be physically attached to the kit. Instead, the user may be directed to an Internet website designated by the manufacturer or seller of the kit.

[0079] Definitions All terms used in this specification are for the purpose of describing particular embodiments only and are not intended to be limiting in any way or scope. For example, as used in this specification and the appended claims, the singular forms "a", "an", and "the" may include plural referents unless the context clearly dictates otherwise. The terms "comprising", "including", and "having" are intended to be inclusive and mean that additional elements other than the recited elements may exist. Further, all units, prefixes, and symbols may be expressed in the SI accepted form.

[0080] As used herein, unless otherwise specified, the following definitions shall apply. For the purposes of the present invention, chemical elements are identified according to the Periodic Table, CAS version, and Handbook of Chemistry and Physics, 75th Ed. 1994. In addition, general principles of organic chemistry are described in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March’s Advanced Organic Chemistry," 5th Ed., Smith, M.B. and March, J., eds. John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

[0081] As used herein, the term "mmol" is intended to mean millimole. As used herein, the term "equiv" is intended to mean equivalent. As used herein, the term "mL" is intended to mean milliliter. As used herein, the term "g" is intended to mean gram. As used herein, the term "kg" is intended to mean kilogram. As used herein, the term "μg" is intended to mean microgram. As used herein, the term "h" is intended to mean hour. As used herein, the term "min" is intended to mean minute. As used herein, the term "M" is intended to mean mole. As used herein, the term "μL" is intended to mean microliter. As used herein, the term "μM" is intended to mean micromole. As used herein, the term "nM" is intended to mean nanomole. As used herein, the term "N" is intended to mean normal. As used herein, the term "amu" is intended to mean atomic mass unit. As used herein, the term "°C" is intended to mean Celsius temperature. As used herein, the term "wt / wt" is intended to mean weight / weight. As used herein, the term "v / v" is intended to mean volume / volume. As used herein, the term "MS" is intended to mean mass spectrometry. As used herein, the term "HPLC" is intended to mean high performance liquid chromatography. As used herein, the term "RT" is intended to mean room temperature. As used herein, the term "for example" is intended to mean example. As used herein, the term "not applicable" is intended to mean not subject to testing.

[0082] The numerical ranges recited within this specification are inclusive of the numbers defining the ranges and include each integer within the defined ranges. Throughout this disclosure, various aspects of the invention are presented in range format. The description in range format is for convenience and brevity only and should not be construed as an invariant limitation on the scope of the invention. Accordingly, a recitation of a range should be considered to specifically disclose all the possible sub-ranges, fractions, and individual numerical values within that range. For example, a recitation of a range such as 1 to 6 should be considered to specifically disclose sub-ranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, as well as the individual numbers within that range, such as 1, 2, 3, 4, 5, and 6, and decimal and fractional numbers, such as 1.2, 3.8, 1 1 / 2, and 4 3 / 4. This applies regardless of the width of the range. In the context of this application, the term "at least" includes the starting point of an open interval. For example, an amount of "at least 95 wt%" means that any amount is equal to or greater than 95 wt%.

[0083] The terms "weight percent", "wt-%", "weight ratio", "weight %", and variations thereof, as used herein, refer to the concentration of a substance obtained by dividing the weight of the substance by the total weight of the composition and multiplying by 100. As used herein, it is understood that terms such as "percent", "%", etc. are intended to be synonymous with "weight percent", "wt-%", etc.

[0084] The term "about", as used herein, refers to variations in quantity that can occur by ordinary measurement techniques and equipment with respect to any quantifiable variable, including but not limited to, for example, mass and volume. Further, when using the handling procedures for given solids and liquids in the real world, there are any inadvertent errors and variations due to differences in the manufacture, source, or purity of the components used in the manufacture of the composition or the implementation of the method, etc.

[0085] The methods and compositions of the present invention may include, consist essentially of, or consist of the components and ingredients of the present invention and other ingredients described herein. As used herein, "consisting essentially of" means that a method and composition may include additional steps, components, or ingredients only if the additional steps, components, or ingredients do not materially alter the basic and novel characteristics of the claimed methods and compositions.

[0086] Without departing from the scope of the present invention, various changes can be made to the above-described compositions and methods. Therefore, all matters included in the above description and the examples shown below are intended to be construed as illustrative rather than in a limiting sense.

Examples

[0087] To demonstrate various embodiments of the present disclosure, the following examples are described. The techniques disclosed in the examples shown below represent techniques that the inventors have discovered to function well in the practice of the present invention and can therefore be considered to constitute a preferred mode for its practice. However, those skilled in the art should understand that, in light of the present disclosure, numerous changes can be made to the disclosed specific embodiments and still obtain the same or similar results without departing from the spirit and scope of the present invention.

[0088] Example 1. Experimental evidence regarding the effect of a 3.6 g mixture of EAA and arginine. The following tests were conducted to quantify the response to two doses of the following amino acid formulations: · 1 - 2% histidine · 9 - 11% isoleucine · 35 - 38% leucine · 14 - 17% lysine · 2 - 4% methionine · 5 - 7% phenylalanine · 8 - 9% threonine · 9 - 11% valine · Tryptophan from 0.002 to 0.8% · Arginine from 8 to 11%

[0089] For the elderly target group (see Table 1), the acute stimulation of muscle protein synthesis caused by the intake of either 3.6 g or 10.8 g of the above amino acid preparation was measured. Based on both cost and flavor, the 3.6 g dose was selected because this dose is commercially feasible. The high dose not only stimulates the FSR of muscle protein but also has been shown to have advantages in long-term outcomes in terms of strength and physical function (13), so the response to 3.6 g was compared with the 10.8 g dose. Muscle protein synthesis was measured before and after the oral intake of the amino acid preparation. Table 1. Characteristics of the subjects

Table 1

[0090] The subjects were tested after an overnight fast. The test procedure required a 5-hour fasting period and 3 hours after the intake of the EAA preparation. Throughout the experiment, a constant infusion of trace 2 H5-phenylalanine was administered to the subjects. Muscle biopsies were taken 2 hours and 5 hours after the trace infusion during the baseline period, and 3 hours after the intake of the amino acid preparation. The concentrations of free phenylalanine and protein-bound phenylalanine in the muscle were measured in each biopsy sample, and the fractional synthesis rate (FSR) of muscle protein over 3 hours at baseline and after the preparation intake was calculated. The FSR of muscle protein represents the proportion of the total muscle mass produced per unit of time (per hour in this experiment). In a short period like this experiment, since the amount of muscle protein is unlikely to change from the start to the end of the test, the FSR of muscle protein directly represents the absolute rate of muscle protein synthesis. The values were calculated as post-value - pre-value. Since various studies have shown that muscle protein synthesis during fasting does not change within the time frame of this test (e.g., 14), each subject was used as their own control in this method. The post-intake values represent the integrated response over the entire 3 hours after the intake of the amino acid preparation.

[0091] After ingestion of the 3.6 g dose, the FSR of muscle increased by 0.0568% / hour. This was a 57% increase over the baseline FSR. The corresponding value was an increase of 0.0692% / hour, a 76% increase over the baseline value. Plasma EAA concentrations were also measured (Table 2). Table 2 Amino acid concentrations (mol / L / 3 hours) *

Table 2

[0092] Three amino acid amounts were administered at high doses, but the increase in muscle protein FSR at the high dose was only slightly greater than the response to the 3.6 g dose. The reason for the overly large response to the low dose is partially explained by the fact that the amino acid concentrations shown in Table 2 do not reflect the degree of difference in the amount of amino acids ingested. This probably indicates that when provided at high doses, EAA oxidation is accelerated, so that a given dose is not utilized as efficiently. In other words, the muscle-level signal to increase synthesis was overly large in the response to the low dose. It also indicates that a 3.6 g dose was sufficient to activate the synthetic reaction in muscle. Another likely explanation for the overly large response to the 3.6 g dose is that the amount of leucine in the 3.6 g formulation was sufficient to induce maximal activation of the initiating factor, and that the increase in FSR at the high dose was relatively small, representing an overall increase in the supply of precursors for synthesis.

[0093] The newly synthesized protein contains all 20 amino acids, but only 9 amino acids are contained in the formulation. The remaining amino acids are derived from more efficient amino acid recycling resulting from proteolysis. Thus, some amino acids, including alanine, are usually released from muscle as a major means, thereby transporting nitrogen to the liver, where it is taken up and converted to ammonia and urea, and finally excreted in urine, but are recycled for protein synthesis instead. Consistent with this interpretation, a decrease in alanine concentration has been observed in previous studies after ingestion of an EAA mixture (15), and enhanced recycling has been shown during the process of protein synthesis. As a result, in contrast to ingestion of an amino acid mixture containing alanine and other NEAA, an increase in urea production was not observed after EAA ingestion (15).

[0094] This optimal ratio of EAA and arginine stimulates muscle protein synthesis at low doses, and this result is unexpected. Typical proteins containing amino acids have effects far lower than 3.6 grams of this composition and would require doses well in excess of 20 grams.

Claims

1. 1. A powdered nutritional formulation for stimulating muscle protein synthesis in a subject, comprising an amino acid composition containing, in w / w %, the following concentrations of amino acids: 1-2% histidine, 9-11% isoleucine, 35-38% leucine, 14-17% lysine, 2-4% methionine, 5-7% phenylalanine, 8-9% threonine, 9-11% valine, 0.005-0.8% tryptophan, and 8-11% arginine; A powdered nutritional formulation, wherein the nutritional formulation is used in a dosage containing between 3 grams and 4 grams of an amino acid composition.

2. 10. The powdered nutritional formula of claim 1, wherein the nutritional formula is added to a consumable liquid to form a nutritional drink.

3. 10. The powdered nutritional formula of claim 1, wherein the nutritional formula is added to a food product.

4. 3. The powdered nutritional formula of any one of claims 1 and 2, wherein the nutritional formula or nutritional drink is packaged in a single or multi-dose container.

5. 5. The powdered nutritional formulation of claim 4, wherein the container is a package, box, carton, wrapper, bottle, or can.

6. 1. A nutritional drink for stimulating muscle protein synthesis in a subject, comprising an amino acid composition containing the following concentrations of amino acids in w / w % terms: 1-2% histidine, 9-11% isoleucine, 35-38% leucine, 14-17% lysine, 2-4% methionine, 5-7% phenylalanine, 8-9% threonine, 9-11% valine, 0.005-0.8% tryptophan, and 8-11% arginine; 10. A nutritional drink, wherein the nutritional drink comprises between 3 grams and 4 grams of an amino acid composition.

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