WO2012024611A1

WO2012024611A1 - Methods of treating sarcopenia and frailty

Info

Publication number: WO2012024611A1
Application number: PCT/US2011/048465
Authority: WO
Inventors: Roger Harris; John Wise
Original assignee: Natural Alternatives International, Inc.
Priority date: 2010-08-20
Filing date: 2011-08-19
Publication date: 2012-02-23

Abstract

Provided are methods that include administration of beta-alanine for the treatment of muscular disorders and for the improvement of muscle function. Among the methods provided are methods for maintaining muscle strength and function (e.g., in the elderly); reversal or prevention of frailty or age-related functional decline in the elderly, reversal or prevention of sarcopenia; and treatment of musculoskeletal impairment in the elderly.

Description

METHODS OF TREATING SARCOPENIA AND FRAILTY

Related Applications

This application claims priority to U.S. Provisional Patent Application No.

61/401,919, filed August 20, 2010, and U.S. Provisional Patent Application No. 61/503,818, filed July 1 , 2011 ; the contents of which are incorporated herein by reference in their entireties.

Field

Provided are methods that include administration of beta-alanine for the treatment of muscular disorders and for the improvement of muscle function. Among the methods provided herein are methods for maintaining muscle strength and function (e.g., in the elderly); reversal or prevention of frailty or age-related functional decline in the elderly, reversal or prevention of sarcopenia; and treatment of musculoskeletal impairment in the elderly.

Background

Worldwide, the number of people 65 or older is increasing. According to a 2010

Administration on Aging report, the population 65 and over in the United States of America will increase from about 40 million in 2010 to about 72.1 million by 2030 (Administration on Aging, A profile of older Americans- 2009, Washington, DC, U. S. Department of Health and Human Services (2010), page 5). It is estimated that by 2050, almost one-third of Europeans will be over 65 years old (Rand Corporation, Does Europe have enough babies! (2010)). In Australia, the population of older adults is expected to constitute about 25% of the population in 2050 (Taaffe, Australian Family Physician, 35(3): 130-133 (2006)). China's 65-and-older population is expected to triple by 2050 and the population of people 65 and over in India is expected to quadruple, with estimates that people 65 or older will constitute 20 percent or more of their population (Aulova, CBS News Political Hotsheet, Census Bureau, Older Population to Triple by 2050 (2009)).

As a consequence of aging, many individuals experience an increased risk of debilitating conditions, diseases or disorders. One of the most prominent and consistent physiological changes that occurs with aging is a loss of muscle mass and strength or function. This progressive decrease in skeletal muscle mass and strength is known as sarcopenia, and contributes to frailty and falls in the elderly. Sarcopenia does not require an underlying disease for manifestation. Decrease in muscle use can result in muscle function loss and/or loss in muscle mass. Sarcopenia has been correlated to functional impairment, disability, falls, frailty, and the loss of independence that increases with aging. The etiology of sarcopenia includes decreased physical activity and can be accompanied by malnutrition or inadequate protein consumption. Loss of muscle function and mass can lead to age-related decline and the onset of frailty. Underlying symptoms of frailty include the progressive loss of robust function in multiple tissues and organ systems, and can lead to decreased muscular support of skeletal structure.

Sarcopenia is a progressive process that occurs throughout adult life, and depending on one's physical activity, by the time a person is over 75 years old, muscle mass may have declined by as much as 50% compared to the amount of muscle mass present in the early twenties. This reduction of muscle mass, in conjunction with loss of muscle functionality, is a significant factor in the development of frailty, which is accompanied by falls that lead to fractures and ultimately to morbidity and mortality. Sarcopenia has been identified as a cause of age-related disability (Harris, J. Nutr. 127: 1004S-1006S (1997); Lexell, J. Gerontol. 50A: 11-16 (1995)). Sarcopenia is believed to be associated with metabolic, physiologic, and functional impairments and disability.

There are few interventions available to treat or prevent sarcopenia and/or frailty.

Anabolic interventions for the treatment of sarcopenia have been suggested. For example, because hormone levels decrease with aging, it has been suggested that hormone

replacement, particularly testosterone or growth hormone supplementation, may be an effective intervention in the treatment of sarcopenia and/or frailty (Bross et ah, J Clin Endocrinology & Metabolism 84(10): 3420-3430 (1999)). Although studies suggest that anabolic steroids may increase lean body mass in older men, no evidence has been shown that such interventions result in any improvements in muscle function, physical performance or reversal of symptoms of sarcopenia or frailty.

In view of the substantially increasing age of the population in the nations of the world, and the inevitability of sarcopenia and age related frailty, it is highly desirable to find more effective methods for treating sarcopenia and/or frailty. Thus, there is a need for more effective methods to maintain muscle strength and function, particularly in the elderly, and to reverse or prevent frailty or age-related functional decline of muscle strength and/or function and/or muscle mass.

Summary

Accordingly, provided herein are methods to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia in a subject. Also provided are methods to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of frailty in a subject. In the methods provided herein, beta-alanine is provided in an amount and over a period of time sufficient to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia and/or frailty in a subject.

Also provided herein are methods for maintaining or increasing muscle strength and/or function and/or muscle mass in a subject having or susceptible of developing sarcopenia or frailty. Also provided are methods of stopping or reversing a decline in skeletal muscle tissue function in a subject having or at risk of developing sarcopenia or frailty.

Also provided are methods of inhibiting muscle catabolism and/or increasing muscle anabolism in a subject having or at risk of developing sarcopenia or frailty. The method includes administering to the subject a beta-alanine in an effective amount and for sufficient time to inhibit muscle catabolism and/or increasing muscle anabolism in the subject. The methods also can include co-administering a protein and/or one or more essential amino acids.

Also provided are methods of improving the muscle:fat ratio in a subject having or at risk of developing sarcopenia or frailty. The method includes administering to the subject a beta-alanine in an effective amount and for sufficient time to improve the muscle: fat ratio in the subject. The methods can include co-administering a protein and/or one or more essential amino acids. The methods also can include modifying the diet of the subject, such as by increasing protein intake and/or reducing high glycemic index carbohydrate intake.

Also provided are methods of improving the gait of a subject having or at risk of developing sarcopenia or frailty. The method includes administering to the subject a beta- alanine in an effective amount and for sufficient time to improve the gait of the subject. The methods can result in an increased stride length, reduced stride frequency, reduced stance width variability or a combination thereof.

Also provided are methods of improving muscle functionality of a subject having or at risk of developing sarcopenia or frailty. The method includes administering to the subject a beta-alanine in an effective amount and for sufficient time to improve the muscle

functionality of the subject. The methods can result in improvements in functionally important tasks, such as improved results in the timed get-up-and-go test, the timed stand test, the stair climb muscle power test, and improvements in balance, such as the one-leg balance test or improvements in one or more criteria of the Berg balance test. Also provided are methods of improving a Berg Balance test score in an elderly subject, the method including administering to the subject a beta-alanine in an effective amount and for sufficient time to improve the Berg Balance test score of the subject compared to a baseline score prior to administration of the beta-alanine. In the methods provided herein, the Berg Balance test score is improved by at least +5, and can be improved by +6, +7, +8, +9, +10, +11, +12, +13, +14, +15 or more. In some methods, the score is improved to be in a range between 30 and 40 or between 40 and 50.

For any of the methods provided herein, the age of the subject can be selected from between 40 and 70 years old or older, such as at least 40, or at least 50, or at least 55, or at least 60, or at least 65, or at least 70 years of age. In some embodiments, a subject is within a particular range of ages. For example, the subject can be selected from among (a) 40-50, (b) 50-60, (c) 60-70 and (d) greater than 70 years of age.

For any of the methods provided herein, the subject may have suffered some loss of muscle mass, but may not suffer from a condition that interferes with acts of daily living and/or prevents the subject from living an independent life. In some embodiments, the subject has suffered some loss of muscle mass and suffers from a condition that interferes with acts of daily living and/or prevents the subject from living an independent life. In some methods, the subject exhibits one or more symptoms of sarcopenia. In some methods, the subject exhibits one or more symptoms of frailty.

For any of the methods provided herein, the beta-alanine can be administered to the subject for any length of time effective to achieve a desired result. For example, the beta- alanine can be administered to the subject for any length of time effective to achieve an increase in the level of a beta-alanyl-histidine dipeptide, such as carnosine, anserine, and balenine or salts or alkyl derivatives of these, particularly carnosine, in a muscle tissue. The beta-alanine can be administered to the subject for any length of time effective to increase the plasma levels of beta-alanine in a subject. In some methods, the beta-alanine can be administered over a period of time of between 1 week and 1 year. In some methods, the beta- alanine can be administered to the subject for at least 6, 8, 10, 12, 14, 16, 18 or 20 weeks. In some methods, the beta-alanine can be administered to the subject for 1 year or more.

For any of the methods provided herein, the beta-alanine can be administered to the subject in any dosage effective to achieve a desired result. For example, the beta-alanine can be administered to the subject in any dosage effective to achieve an increase in the level of a beta-alanyl-histidine dipeptide, such as carnosine in a muscle tissue. The beta-alanine can be administered to the subject in any dosage effective to increase the plasma levels of beta- alanine in a subject. In some embodiments, the beta-alanine can be administered in a dose of between 0.1 g and 16 g per day. In some embodiments, the beta-alanine can be administered as a dosage of 200 mg, 400 mg, 600 mg, 800 mg, 1,000 mg or 1,200 mg, 1,400 mg, 1,600 mg, 1,800 mg or 2,000 mg one or more times a day. Any dosage form for administering the beta-alanine can be used in the methods provided herein. For example, the beta-alanine can be administered as a bolus dosage or as a sustained release or controlled release dosage or a combination of immediate release and controlled release forms. The dosage forms can be formulated to deliver an amount of beta-alanine in a 24 hour period that is between about 0.2 grams and 20 grams. In some methods, a dosage of beta-alanine delivered over a 24 hour period is greater than 1 gram. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 1.6 grams or more. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 2.4 grams or more. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 3.2 grams or more. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 4.0 grams or more. In some methods, a dosage of beta- alanine delivered over a 24 hour period is 4.8 grams or more. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 5.6 grams or more. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 6.4 grams or more. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 7.2 grams or more. In some methods, a dosage of beta-alanine delivered over a 24 hour period is 8.0 grains or more.

In any of the methods provided herein, the methods can include co-administering a protein and/or one or more essential amino acids. The methods also can include modifying the diet of the subject, such as by increasing protein intake and/or reducing high glycemic index carbohydrate intake. The methods also can include co-administering one or more vitamin and/or mineral supplement. In some embodiments, the methods include

administering vitamin D or an analog thereof.

In any of the methods provided herein, the methods can include exercise as a step. The exercise can be aerobic exercise or anaerobic exercise or a combination thereof. In some embodiments, the methods include administering beta-alanine prior to the exercise. In some embodiments, the methods include administering beta-alanine after the exercise. In some of the methods provided herein, the methods include resistance exercise as a step.

In any of the methods provided herein, the beta-alanine can be administered in a dosage effective to maintain a plasma concentration of beta-alanine above the minimal effective concentration (MEC). In some of the methods, the dosage of beta-alanine is effective to maintain a plasma concentration of beta-alanine above the MEC for 10-90% of the time, or between 30-90% of the time, or between 50-90% of the time or between 20-80% of the time.

In any of the methods provided herein, the beta-alanine can be administered at a dosage and over a period of time sufficient to increase muscle carnosine levels at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100%, or at least 110%, or at least 120% , or at least 130% , or at least 140% , or at least 150% or more from the level before

administration of initial beta-alanine commenced.

In any of the methods provided herein, the beta-alanine can be administered at a dosage and over a period of time sufficient to increase muscle carnosine levels to at least 30 mmol/kg dry muscle weight, or at least 40 mmol/kg dry muscle weight, or at least 50 mmol/kg dry muscle weight, or at least 60 mmol/kg dry muscle weight.

These and other aspects, which will become apparent during the following detailed description, have been achieved by the discovery that beta-alanine can be used to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia or frailty in a subject. Brief Description of the Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate preferred embodiments of the invention and together with the detailed description serve to explain the principles of the invention. In the drawings:

Fig. 1 are graphs showing individual data for muscle carnosine content (arbitrary units) at baseline (PRE) and after 12 weeks of beta-alanine supplementation following either beta-alanine (Panel A, n = 10) or placebo supplementation (Panel B, n = 6).

Fig. 2 is a graph showing individual data for absolute change in muscle carnosine content (arbitrary units) from baseline (PRE) to 12-week supplementation period (POST- 12).

Fig. 3 is a graph showing time-to-exhaustion in the submaximal exercise test (TLIM; i.e., intensity corresponding to 75% of the difference between ventilatory threshold and V02 peak) at baseline (PRE) and after 12 weeks of beta-alanine supplementation (POST-12). Fig. 4 is a graph showing correlation between percent change in the time-to- exhaustion in the TLIM test (i.e., intensity corresponding to 75% of the difference between ventilatory threshold and V02 peak) and the percent change in the muscle carnosine content.

Fig. 5 is a graph showing an absolute change in time-to-exhaustion in the incremental test at baseline (PRE) and after 12 weeks of beta-alanine supplementation (POST-12). * denotes p = 0.06 or p = 0.04, with or without subject 6.

Fig. 6 is a graph showing correlation between percent change in the time-to- exhaustion in the incremental test and the percent change in the muscle carnosine content. Detailed Description

For clarity of disclosure, and not by way of limitation, the detailed description is divided into subsections that follow.

A. Definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the inventions belong. All patents, patent applications, published applications and publications, websites and other published materials referred to throughout the entire disclosure herein, unless noted otherwise, are incorporated by reference in their entirety. In the event that there are a plurality of definitions for terms herein, those in this section prevail. Where reference is made to a URL or other such identifier or address, it understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

As used herein, the term "active agent" refers to a drug or any compound that is a therapeutic agent or a candidate for use as a therapeutic or as lead compound for designing a therapeutic or that is a known pharmaceutical. Such compounds can be small molecules, including small organic molecules, peptides, peptide mimetics, antisense molecules, antibodies, fragments of antibodies, recombinant antibodies.

As used herein, "biological activity" refers to the in vivo activities of a compound or physiological responses that result upon in vivo administration of a compound, composition or other mixture. Biological activity, thus, encompasses therapeutic effects and

pharmaceutical activity of such compounds, compositions and mixtures. Biological activities can be observed in in vitro systems designed to test or use such activities. As used herein, the term "physical capacity" refers to a measure of the ability of active muscle systems to deliver, by aerobic metabolism or anaerobic metabolism, energy for mechanical work, and to continue working for as long as possible.

As used herein, the term "assess" and grammatical variations thereof, are intended to include quantitative and qualitative determination in the sense of obtaining an absolute value for the activity of a polypeptide, and also of obtaining an index, ratio, percentage, visual or other value indicative of the level of the activity. Assessment can be direct or indirect.

As used herein, the term "contacting" refers to bringing two or more materials into close enough proximity whereby they can interact. In certain embodiments, contacting can be accomplished in a vessel such as a test tube, a Petri dish, or the like. In certain embodiments, contacting can be performed in the presence of additional materials. In certain embodiments, contacting can be performed in the presence of cells. In certain of such embodiments, one or more of the materials that are being contacted can be inside a cell. Cells can be alive or can be dead. Cells can or can not be intact.

As used herein, a "combination" refers to any association between two or among more items. The association can be spatial or refer to the use of the two or more items for a common purpose.

As used herein, a "composition" refers to any mixture of two or more products or compounds (e.g., agents, modulators, regulators, etc.). It can be a solution, a suspension, liquid, powder, a paste, aqueous or non-aqueous formulations or any combination thereof.

As used here, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise.

As used herein, ranges and amounts can be expressed as "about" a particular value or range. "About" is intended to also include the exact amount. Hence "about 5 percent" means "about 5 percent" and also "5 percent." "About" means within typical experimental error for the application or purpose intended.

As used herein, "optional" or "optionally" means that the subsequently described event or circumstance does or does not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

As used herein, the term "pharmaceutically acceptable salt" is intended to include all salts known and used in the art of pharmaceuticals. Pharmaceutically acceptable salts include, but are not limited to, amine salts, such as but not limited to chloroprocaine, choline, Ν,Ν'- dibenzyl-ethylenediamine, ammonia, diethanolamine and other hydroxyalkylamines, ethylenediamine, N-methylglucamine, procaine, N-benzyl phenethylamine, 1-para-chloro- benzyl-2-pyrrolidin- -ylmethyl-benzimidazole, diethylamine and other alkylamines, piperazine and tris(hydroxy-methyl)aminomethane; alkali metal salts, such as but not limited to lithium, potassium and sodium; alkali earth metal salts, such as but not limited to barium, calcium and magnesium; transition metal salts, such as but not limited to zinc; and other metal salts, such as but not limited to sodium hydrogen phosphate and disodium phosphate; and also including, but not limited to, salts of mineral acids, such as but not limited to hydrochlorides and sulfates; and salts of organic acids, such as but not limited to acetates, lactates, malates, tartrates, citrates, ascorbates, succinates, butyrates, valerates and fumarates. Exemplary pharmaceutically acceptable salts include acetate, lactobionate, benzenesulfonate, laurate, benzoate, malate, bicarbonate, maleate, bisulfate, mandelate, bitartrate, mesylate, borate, methylbromide, methylnitrate, calcium edetate, methylsulfate, camsylate, mucate, carbonate, napsylate, bromide, chloride, nitrate, clavulanate, N-methylglucamine, citrate, ammonium salt, dihydrochloride, oleate, edetate, oxalate, edisylate, pamoate (embonate), estolate, palmitate, esylate, pantothenate, fumarate, phosphate/ diphosphate, gluceptate, polygalacturonate, gluconate, salicylate, glutamate, stearate, glycollylarsanilate, sulfate, hexylresorcinate, subacetate, hydrabamine, succinate, hydrobromide, tannate, hydrochloride, tartrate, hydroxynaphthoate, teoclate, iodide, tosylate, isothionate, triethiodide, lactate, panoate and valerate, which can be used as a dosage form for modifying the solubility or hydrolysis characteristics or can be used in sustained release or pro-drug formulations. The preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts is more fully described by Berg et ah, "Pharmaceutical Salts," J. Pharm. Sci. 66: 1-19 (1977).

Pharmaceutically acceptable esters include, but are not limited to, alkyl, alkenyl, alkynyl, cycloalkyl and heterocyclo esters of acidic groups, including, but not limited to, carboxylic acids, phosphoric acids, phosphinic acids, sulfonic acids, sulfinic acids and boronic acids. Pharmaceutically acceptable enol ethers include, but are not limited to, derivatives of formula C=C(OR) where R is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, heteroaralkyl, cycloalkyl or heterocyclyl. Pharmaceutically acceptable enol esters include, but are not limited to, derivatives of formula C=C(OC(0)R) where R is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, heteroaralkyl, cycloalkyl or heterocyclyl. Pharmaceutically acceptable solvates and hydrates are complexes of a compound with one or more solvent or water molecules, or 1 to about or 100, or 1 to about or 10, or one to about or 2, 3 or 4, solvent or water molecules.

Pharmaceutically acceptable salts include alaninate salts, including, but not limited to, creatine-, -alaninate, tauryl-beta-alaninate alone or complexed with one or more metal ions selected from among zinc, manganese, magnesium, calcium, copper, iron, boron, vanadium, molybdenum, germanium and selenium, nickel, vanadium, silicon, germanium, arsenic, aluminum, cadmium, lithium, cobalt and rubidium and aryl-beta-alaninates, such as benzyl- beta-alaninate.

As used herein, the term "functional food" refers to a food that includes potentially healthful products including any modified food or food ingredient that provides a health benefit beyond the traditional nutrients it contains, and can include a nutraceutical. For example, see (Hasler, Functional Foods: the Western perspective, Nutrition Reviews, 54: S6- S10 (1996)).

As used herein, the term "nutraceutical" refers to any substance, agent or combination of agents, that produces a physiological effect in a mammal, such as a medical or health benefit. Nutraceuticals may be derived from natural sources or prepared synthetically.

Representative nutraceuticals include but are not limited to bioflavonoids, catechin-based preparations such as proanthocyanidin, acerola concentrate, grape seed extract, pycnogenol, provatene, carotenoids such as β-carotene, sodium bisulfite, vitamins such as Vitamin E, riboflavin (Vitamin B2), and Vitamin C (L-ascorbic acid), a-tocopherol, all manner of herbal compounds, elderberry extract, lutein, coenzyme Q10, and combinations thereof.

As used herein, the abbreviations for any protective groups, amino acids and other compounds, are, unless indicated otherwise, in accord with their common usage, recognized abbreviations, or the IUPAC-IUB Commission on Biochemical Nomenclature (see, e.g., Biochem. 1 1 : 1726 (1972)).

As used herein, the term "sample" refers to any composition, whether liquid, gas or solid, that includes a molecule or material to be detected or examined. A sample can be water or a buffered solution or be composed of any artificially introduced chemicals, and may or may not contain nucleic acids, amino acids or peptides. The sample can be a biological sample, such as a biological fluid or a biological tissue obtained from any organism or a cell of or from an organism or a viral particle or portions thereof.

As used herein, a "drug" refers to any compound that is a candidate for use as a therapeutic or as lead compound for designing a therapeutic or that is a known pharmaceutical. Such compounds can be small molecules, including small organic molecules, peptides, peptide mimetics, antisense molecules, antibodies, fragments of antibodies, recombinant antibodies.

As used herein, "derivative" or "analog" of a molecule refers to a portion derived from or a modified version of the molecule.

As used herein, a "therapeutic agent" or "therapeutic regimen" refers to conventional drugs and drug therapies, which are known to those skilled in the art, and includes compounds that exhibit a therapeutic effect when administered to a subject.

As used herein, the term "C_mx" refers to the maximum (peak) observed plasma concentration.

As used herein, the term "T_max" refers to the time to reach the maximum (peak) observed plasma concentration C_mx.

As used herein, "fluid" refers to any composition that can flow. Fluids thus encompass compositions that are in the form of semi-solids, pastes, solutions, aqueous mixtures, gels, lotions, creams and other such compositions.

As used herein, the term "inhibit" refers to the ability of a compound to reduce or impede a described function.

As used herein, the term "standard" refers to something used for comparison. For example, a standard can be a known standard agent or compound that is administered or added to a control sample and used for comparing results when measuring said compound in a test sample. Standard can also refer to an "internal standard," such as an agent or compound which is added at known amounts to a sample and is useful in determining such things as purification or recovery rates when a sample is processed or subjected to purification or extraction procedures before a marker of interest is measured.

As used herein, the term "treat" or "treatment" refers to an action resulting in a curative treatment or to a lessening or reduction in the severity of a symptom of or a disease or condition or to lessening the frequency of outbreaks of a disease or disorder. The terms include a remitative treatment of a disorder (i.e. treatment that causes the disorder to enter remission). The term "treat" or "treatment" includes administration of an agent where the disease or condition is at least partially improved or ameliorated, and/or there is some alleviation, mitigation or decrease in at least one clinical symptom, and/or there is a delay in the progression of the condition or disease, and/or prevention or delay of the onset of the condition or disease. Thus, the terms "treat" and "treatment" refer to both prophylactic and therapeutic treatment regimes.

As used herein, prevention or prophylaxis refers to methods in which the risk of developing disease or condition is reduced. Prophylaxis includes reduction in the risk of developing a disease or condition and/or a prevention of worsening of symptoms or progression of a disease or reduction in the risk of worsening of symptoms or progression of a disease or condition.

The terms "reducing", "suppressing" and "inhibiting" refer to lessening or decreasing. As used herein, an "effective amount" of a compound or composition for treating a particular condition or disease is an amount that is sufficient to ameliorate, or in some manner reduce the symptoms associated with the condition or disease. Such amount can be administered as a single dosage or can be administered according to a regimen, whereby it is effective. The amount can cure a disease, but, typically, can be administered in order to ameliorate the symptoms of the condition or disease, or to prevent the onset of the condition or disease. Typically, repeated administration is required to achieve a desired amelioration of symptoms.

As used herein, the term "therapeutically effective amount" or "therapeutically effective dose" refers to an amount of an agent, such as a beta-alanine, that is at least sufficient to produce a therapeutic effect. An effective amount is the quantity of a therapeutic agent necessary for preventing, curing, ameliorating, arresting or partially arresting a symptom of a disease, condition or disorder.

As used herein, the term "ameliorate" or the term "amelioration of the symptoms" of a particular condition, disease or disorder by administration of a particular compound or pharmaceutical composition that includes the compound refers to any lessening of severity, delay in onset, slowing of progression, or shortening of duration, whether permanent or temporary, lasting or transient, that can be attributed to or associated with administration of the compound or composition containing the compound. As used herein, the term "prevent" means that a subject does not present the phenotypical symptoms of the disease or condition within the time during which a subject not exposed to the treatment agent, such as beta-alanine, would be expected to develop traits characteristic of the particular disease or condition.

As used herein, the term "mitigate" refers to a decrease in the severity of traits or symptoms of a disease or condition. Mitigation can be quantitated, such as using the methods and parameters described herein or known in the art, and mitigation includes a decrease in the severity of traits or symptoms of a disease or condition of at least 10% compared to subject, equally disposed to develop a particular disease or condition, which has not been exposed to the treatment agent.

As used herein, the term "onset" refers to the beginning of detectable traits or symptoms of a disease or condition.

As used herein, the term "decrease," e.g., when referring to a decrease in the severity of a disease or condition, refers to a lessening of one or more symptoms of the disease or condition when comparing symptom severity in subjects treated with an agent, such as beta- alanine, to symptom severity in subjects treated with a placebo or to subjects not treated. In certain embodiments, the lessening or decrease is statistically significant, e.g., having a P < 0.05.

As used herein, the term "administering" refers, in one embodiment, to bringing a subject in contact with beta-alanine, a salt of beta-alanine, a derivative of beta-alanine or a compound comprising beta-alanine, such as a peptide that includes beta-alanine.

As used herein, "frailty" refers to an adverse, primarily gerontologic, health condition, which can include low functional reserve, accelerated osteoporosis, easy tiring, decreased muscle strength, high susceptibility to disease and decreased libido (e.g., see Bandeen-Roche et ah, The Journals of Gerontology Series A: Biological Sciences and Medical Sciences 61 : 262-266 (2006)). Frailty can be characterized by meeting three of the following five attributes: unintentional weight loss, muscle weakness, slow walking speed, exhaustion, and low physical activity.

As used herein, "sarcopenia" means a loss of skeletal muscle mass, quality, and strength. Sarcopenia may lead to frailty, for example, in the elderly.

As used herein, the term "subject" refers to an animal, such as a mammal, for example a human, that has been or will be the object of treatment, observation or experiment. The methods described herein can be useful in both human therapy and veterinary applications. In some embodiments, the subject is a mammal, and in some embodiments, the subject is human. In some embodiments, the subject is a companion animal.

As used herein, "animal" includes any animal, such as, but not limited to; primates including humans, gorillas and monkeys; rodents, such as mice and rats; fowl, such as chickens; ruminants, such as goats, cows, deer, sheep; ovine, such as pigs and other animals. Non-human animals exclude humans as the contemplated animal.

As used herein, the term "mammal" refers to any animal classified as such, including humans, domestic and farm animals, zoo, sports, or pet animals, such as dogs, horses, cats, sheep, pigs, cows, etc.

As used herein, the term "improvement of muscle function" encompasses the enhancement of the physical performance especially the enhancement of the physical endurance and the fatigue resistance.

As used herein, the phrase "inhibiting the development" of a sign of aging means delaying the onset, slowing the progression, or reducing the manifestation, of a sign of aging.

As used herein, the term "improving performance" refers to any aspect of performance, including cognitive performance or physical performance, such as, but not limited to, the ability to be self-sufficient, to take care of (some but not necessarily all) personal needs, to be ambulatory or otherwise mobile, or interaction with others.

As used herein, the term "muscle strength" refers to the ability of a muscle or a group of muscles to produce tension or exert force through the skeletal system.

As used herein, "skeletal muscle" includes skeletal muscle tissue as well as components thereof, such as skeletal muscle fibers (i.e., fast or slow skeletal muscle fibers), the myofibrils comprising the skeletal muscle fibers, the skeletal sarcomere which comprises the myofibrils, and the various components of the skeletal sarcomere described above.

Skeletal muscle does not include cardiac muscle or a combination of sarcomeric components that occurs in such combination only in cardiac muscle.

As used herein, "power output" of a muscle means work/cycle time. Power output may be modulated by changing, for example, activating parameters during cyclical length changes, including timing of activation (phase of activation) and the period of activation (duty cycle).

As used herein, the term "muscle function" refers to any one or more physical attributes which can be dependent to any degree on skeletal muscle contraction. For example, muscle functions include, but are not limited to, maximal muscular strength, muscular endurance, running speed and endurance, swimming speed and endurance, throwing power, lifting and pulling power, ability to change position (such as measured by the timed get-up- and-go test), translocation, such as can be evaluated by the timed stand test, and skeletal muscle support and coordination, such as can be measured by a subject's control of balance, such as can be measure by the one-leg balance test (Gillette-Guyonnet et ah, Gerontology 46(4): 189-193 (2000)) or one or more criteria of the Berg balance test.

As used herein, the term "balance" refers to the ability of a subject to maintain the center of gravity over a base of support, usually in an upright position. Balance is complex and is a coordinated response of the neuromuscular and musculoskeletal systems. Decline in muscular function leads to a loss of balance control. With good balance, a patient has the ability to sit, stand, or walk safely without falling or requiring an external means of support.

As used herein, the term "muscle catabolism" refers to muscle degradation, including during intense exercise and prolonged periods of exertion.

As used herein, the term "muscle anabolism" refers to muscle synthesis, generation or regeneration.

As used herein, the term "hand grip strength" refers to the maximum isometric strength of the hand and forearm muscles. Handgrip strength is often used as a general test of strength. Handgrip strength has been demonstrated as a predictor of muscle function, including in the oldest old (e.g., see Taekema et ah, Age Ageing 39(3): 331-337 (2010)). Hand grip strength can be measured by use of a dynamometer or any technique known in the art.

As used herein, the term "at-risk subject" refers to a subject who exhibits objective evidence of decline in muscle performance as measured by established methods of physical performance assessment.

As used herein, the term "appendicular skeletal muscle mass" is the mass of the subject divided by the square of the height of the subject (kg/height² (m²)). Appendicular skeletal muscle mass can be measured by any measurement technique known in the art (e.g., by dual-photon absorptiometry (Heymsfield et ah, Am J Clin Nutr 52: 214-218 (1990)) and DEXA, (Going et ah, Am J Clin Nutr 57: 845-850 (1993), Kellie, JAMA 267: 286-294 (1992), and Roubenoff et ah, Am J Clin Nutr 58: 589-591 (1993)). These techniques permit the body to be segmented into three components: bone, fat, and fat-free soft tissues.

As used herein, the term "appendicular skeletal muscle mass t-score" refers to the standard deviation in appendicular skeletal muscle mass in a subject compared to the mean of a young reference group, such as described in Baumgartner et al. (Am J Epidemiol 147: 755- 763 (1998) and Am J Epidemiol 149: 1161 (1998)). A t-score of -1 refers to an appendicular skeletal muscle mass of a subject one standard deviations below the mean of a young reference group. A t-score of -2 refers to an appendicular skeletal muscle mass of a subject two standard deviations below the mean of a young reference group. Baumgartner et al. suggests that sarcopenia is present in a subject having an appendicular skeletal muscle mass less than two standard deviations below the mean of a young reference group.

As used herein, "cognitive function" refers to any mental component of brain function. For example, cognitive functions include, but are not limited to, attention, concentration, memory and focus.

As used herein, "beta. -alanine" or "β-alanine" refers to the naturally occurring beta amino acid that has the IUPAC name β-amino-propanoic acid (CAS Registry No. 107-95-9).

As used herein, "a beta-alanine" includes free beta-alanine, a biological source of beta-alanine, a salt of beta-alanine or containing beta-alanine, including alaninate salts, and an ester, ether, amide, azide, oxide, hydrate, solvate or chelate of beta-alanine.

As used herein, a compound that is "a biological source of beta-alanine" is a compound that, when administered to the body by any route (for example parenterally, orally, topically), is converted, e.g., via ionic dissolution to constituent ions or by one or more chemical- or enzyme-catalyzed reaction steps, to beta-alanine, which then appears in blood, plasma or serum and is available for uptake into muscle and other tissues.

As used herein, "beta amino acids" are amino acids in which the amino group is at the β-position from the carboxylate group (i.e., two atoms away). Unlike its normal counterpart, L-a-alanine, beta-alanine has no chiral center.

As used herein, "creatine" refers to the chemical N-methyl-N-guanyl glycine, (CAS Registry No. 57-00-1), also known as (alpha-methyl guanido) acetic acid, N-

(aminoiminomethyl)-N-glycine, methylglycocyamine, methylguanidoacetic acid and as N- methyl-N-guanylglycine. Additionally, as used herein, "creatine" also includes glycocyamine (CAS# 352-97-6), guanidinopropionic acid (CAS# 353-09-3), creatinol (CAS# 6903-79-3; FIG. 2), and cyclocreatine (CAS# 35404-50-3), as well as any salt, ester, ether, amide, azide, oxide, or chelate thereof or of creatine.

As used herein, the term "minimal effective concentration" refers to a concentration of beta-alanine required to increase the mean beta-alanylhistidine level in a tissue by at least 10% as compared to that in the absence of administration of beta-alanine. As used herein, the term "enteral nutrition product" refers to a supplemental food material that is provided via the gastrointestinal tract by mouth (orally), or through a tube, catheter, or stoma that delivers nutrients distal to the oral cavity. Enteral nutrition, whether orally or by tube feeding, is used as a therapeutic regimen to prevent serious disability or death in a subject with a condition that precludes the full use of regular food. A variety of such products are available, for example, from Nestle Clinical Nutrition, Abbott, Novartis, Numico, and Fresenius. In an embodiment, these products are provided to the patient outside of a hospital setting. For example, the products can be provided in a nursing home, out care patient center, or even the home of the patient. Any suitable container can be used to supply the nutrition product. Typically, the product is administered so that the patient receives 1500 ml per day, although those skilled in the art will appreciate that variations to the amount of product administered are possible.

As used herein, the term "parenteral nutrition product" refers to a food replacement composition that is administered by means other than through the alimentary tract (as by intramuscular or intravenous injection).

As used herein, "vitamin D" refers to a group of fat-soluble secosteroids, the two major physiologically relevant forms of which are vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol). The term also their precursor molecules such as ergosterol (7-dehydro- 22-dehydro-24-methyl-cholesterol) and 7 dehydrocholesterol, 25-hydroxy-vitamin D 3, the 3- hydroxylated dihydrotachysterol 2, the 1 a-hydroxylated alfacalcidol (1 a-hydroxyvitamin D 3) and calcitriol (1 a, 25-dihydroxyvitamin D 3), as well as the numerous natural and synthetic Vitamin D analogs described in Bouillon et ah, Endocrine Reviews 16: 200-257 (1995).

As used herein, "vitamin D drug" refers to any compound that raises the blood or tissue level of Vitamin D, or has an affinity for the Vitamin D receptor, for example binding to that receptor with a Relative Competitive Index (RCI) of 0.05 or greater. The RCI is indexed to an RCI of 100 for calcitriol. Vitamin D drugs include Vitamin D preparations and analogs, such as Rocaltrol^® (Roche Laboratories), Calcijex^® injectable calcitriol,

investigational drugs from Leo Pharmaceutical including EB 1089 (24a,26a,27a-trihomo- 22,24-diene-la, 25-(OH) 2-D 3), KH 1060 (20-epi-22-oxa-24a,26a,27a-trihomo-l a, 25-(OH) 2-D 3), MC 1288 and MC 903 (calcipotriol), Roche Pharmaceutical drugs that include 1,25- (OH) 2-16-ene-D 3, l,25-(OH) 2-16-ene-23-yne D 3, and 25-(OH)2-16-ene-23-yne-D 3, Chugai Pharmaceuticals 22-oxacalcitriol (22 oxa-la,25-(OH) 2-D 3; la-(OH)D 5 from the University of Illinois; and drugs from the Institute of Medical Chemistry-Schering AG that include ZK 161422 and ZK 157202.

As used herein, the term "unit dosage forms" refers to physically discrete units suitable for human and animal subjects. Each unit dosage includes a predetermined quantity of the therapeutically active compound sufficient to produce the desired therapeutic effect, in association with, when required, a pharmaceutical carrier, vehicle or diluent. Examples of unit dosage forms include tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, ampoules and syringes, and oral solutions or suspensions, and oil- water emulsions. Unit dosage forms can be individually packaged as is known in the art, such as in blister packs. Unit dosage forms can be administered in fractions or multiples thereof.

As used herein, the term "elderly" refers to an individual who has reached the age of 65 years or older.

As used herein, the term "geriatric" refers generally to an adult individual who has reached old age.

As used herein, the term "improving gait" or "improvement in gait" refers to one or more of an increase in stride length, a decrease or reduction in stride frequency and/or a decrease or reduction in stance width variability and/or a decrease in stride length variability in a subject following administration of a beta-alanine to the subject, relative to the stride length, stride frequency, stance width variability, and/or stride length variability before treatment with beta-alanine.

As used herein, the term "stride length" refers to the distance traveled during one cycle of gait (e.g., the distance traveled between the point at which a foot, paw, knee, hand, etc. of a moving (e.g., ambulating) subject departs contact with a primary supporting surface (e.g., the ground or other walking surface) and the point at which the same foot, paw, knee, hand, etc. of the subject next contacts the supporting surface.

As used herein, the term "standardized average stride length" refers to the average measured value for stride length observed for a population that has not been selected for, or is not anticipated to have been selected for, a disease, disorder or any other attribute that alters, or would be anticipated to alter, the measured average stride length of the population.

As used herein, the term "stride frequency" refers the number of strides taken in a given amount of time or over a given distance. As used herein, the term "ergogenic" refers to the ability to increase capacity for bodily and/or mental labor, especially by reducing or eliminating signs and symptoms of fatigue.

As used herein, the term "ready-to-drink meal replacement beverage" refers to any liquid beverage providing nutrition, including the vitamins and minerals known to be essential for human health, as well as protein, carbohydrate and/or fat, for example, including but not limited to Unilever's Slimfast^®, Abbott's Ensure^®, or Mead Johnson's Boost^® meal replacement beverages.

As used herein, the term "anaerobic" means without oxygen.

As used herein, the term "anaerobic exercise" refers to exercise that does not increase the body's requirement for oxygen. Typically, anaerobic exercise can be a short-burst, higher-intensity exercise. Proteins and carbohydrates may be utilized to build muscle mass and/or strength. Fat burning may be an indirect effect of anaerobic exercise. Anaerobic exercise may include, for example and not by way of limitation, push-ups, pull-ups, sit-ups, sprinting, stomach crunches, weight lifting and strength training.

As used herein, the term "aerobic" means with oxygen.

As used herein, the term "aerobic exercise" refers to exercise that increases the body's requirement for oxygen. Typically, aerobic exercise involves an increased respiratory rate and cardiac rate over an extended period of time. Following approximately 20 minutes of aerobic exercise, the body usually requires the utilization of stored fat deposits as fuel for muscle contraction. Therefore, aerobic exercise can be considered to have a direct fat burning effect. Aerobic exercise may include, for example and not by way of limitation, basketball, bicycling, cross-country skiing, ice hockey, ice skating, jogging, martial arts, rollerblading, rowing, soccer, swimming, tennis and walking (e.g., at a fast pace).

As used herein, the term "resistance exercise" refers to exercise that is performed by a subject against resistance, e.g., as from a weight.

As used herein, the term "V0_2max "or "V0_2peak" refers to the maximal amount of oxygen that can be transported and utilized for energy production during exercise, including during incremental or exhaustive exercise.

can be used to gauge the physical fitness of an individual.

As used herein, the term "functional reserve" refers to the remaining capacity of an organ or body part to fulfill its physiological activity. Functional reserve can be impaired or decline do to disease and/or ageing. As used herein, the term "skeletal muscle tissue function" refers to strength, endurance, or motor control. Improving one of these three variables, alone or in combination, results in improved muscle function. Skeletal muscle tissue function includes the ability of muscle to perform a physiologic function, such as contraction, as measured by the amount of force generated during either twitch or tetanus. Methods for assessing muscle function are well known in the art and include, but are not limited to, measurements of muscle mass, grip strength, serum CK level, activities of daily living, motion or strength tests, tissue histology (e.g., collagen III staining), or tissue imaging.

As used herein, the "Berg balance test" refers to an observational performance-based assessment tool that is used to evaluate an individual's balance function during functional activities. The Berg Balance test is a well-known example of an observational test with documented ability to detect fall risk. The Berg test requires a clinically trained individual to observe and numerically rate an individual's ability to perform a series of standardized balance and movement tasks. This test has the advantage of requiring no specialized equipment. The subject is scored on a combination of different tasks, which include changing position from sitting to standing, changing position from standing to sitting, sitting without support, standing without support, standing with eyes closed, standing with feet together, standing on one leg, tandem standing, turning trunk with feet fixed, turning 360 degrees, retrieving objects from the floor, reaching forward while standing and stepping on a stool. Each of the difference tasks is assigned a numerical value from 0 to 4. Subjects that score 20 or less have poor balance, and may be confined to a wheelchair. Subjects with a score between 21 and 40 generally display some problem maintaining balance and may require assistance while walking. Subjects scoring 41 or above generally can control their balance and can walk independently. On average, subjects with scores less than 40 are much more likely to fall than subjects with scores higher than 40.

As used herein, the term "controlled release coat" refers to a functional coat that can, e.g., include at least one pH independent polymer, pH dependent (such as for example enteric or reverse enteric types) polymer, soluble polymer, insoluble polymer, lipids, lipidic materials or combinations thereof which when applied onto a dosage form can slow (for example when applied to a normal release matrix dosage form), further slow (for example when applied to a controlled release matrix dosage form) or modify the rate of release of the beta-alanine when applied to an uncoated dosage form. For example, the control releasing coat can be designed such that when the control releasing coat is applied to a dosage form, the dosage form in conjunction with the control releasing coat can exhibit the release of the beta-alanine, such as for example, as a "modified-release," "controlled-release," "sustained- release," "extended-release," "delayed-release," "prolonged-release" or combinations thereof. The "control releasing coat" can optionally include additional materials that can alter the functionality of the control releasing coat.

As used herein, the term "co-administer" refers to administering more than one pharmaceutical agent to a subject. In certain embodiments, co-administered pharmaceutical agents are administered together in a single dosage unit. In certain embodiments, coadministered pharmaceutical agents are administered separately. In certain embodiments, co- administered pharmaceutical agents are administered at the same time. In certain

embodiments, co-administered pharmaceutical agents are administered at different times. BETA-ALANINE

Beta-alanine, or 3 -amino-propanoic acid, is formed in vivo by the degradation of dihydrouracil and carnosine, and is readily obtained from meat in a carnivorous or omnivorous diet. Beta-alanine is a component of a number of naturally occurring

beta-alanylhistidine peptides, including carnosine and anserine as well as being a component of pantothenic acid (vitamin B5). Beta-alanine is the rate-limiting precursor of the synthesis of carnosine. Supplementation with beta-alanine has been shown to increase the

concentration of carnosine in muscles and decrease fatigue in athletes (Harris et al, Amino Acids 30: 279-289 (2006); Derave et al, J Appl Physiol 103 : 1736-1743 (2007); Hill et al, Amino Acids 32(2): 225-233 (2007)). It has been shown that increasing muscle carnosine levels increases buffering capacity of the muscle. Because accumulation of lactic acid and/or H⁺ ions and/or hydronium ions in muscle is a cause of rapid muscle fatigue during strenuous exercise, increased muscle buffering capacity can extend the duration of extended exercise before muscle fatigue (e.g., see Hill et al., Amino Acids 32(2): 225-233 (2007)).

Beta-alanine and L-histidine and their methylated analogues form dipeptides within the human or animal body. The di-peptides produced from beta-alanine and histidine include carnosine (beta-alanyl-L-histidine), anserine (beta-alanyl-L-l-methyl-histidine), or balenine (beta-alanyl-L-3-methylhistidine) (referred to collectively herein as beta-alanylhistidine peptides). Beta-alanylhistidine peptides are involved in the regulation of intra-cellular pH homeostasis during muscle contraction and, therefore, are involved in the regulation of muscle fatigue. Beta-alanylhistidine peptides provide an effective way of accumulating pH- sensitive histidine residues in a cell. Thus, variations in the muscle beta-alanylhistidine peptide concentrations affect the anaerobic work capacity of muscles, and increasing the amount of beta-alanylhistidine peptides within a muscle favorably affects performance and the amount of work that can be performed by the muscle.

Beta-alanine and L-histidine can be produced by the body or can be obtained through the diet that includes a meat protein source. Within the body, beta-alanine is transported to tissues such as muscle. Since in a typical fed state, the concentration of beta-alanine in muscle is low in comparison with the concentration of L-histidine, the concentration of beta- alanine is likely limiting to the synthesis of beta-alanylhistidine peptides. The synthesis and accumulation of beta-alanylhistidine peptides in a human or animal body can be increased by increasing the blood or blood plasma concentrations of beta-alanine, increasing the blood or blood plasma concentrations of beta-alanine and creatine, or increasing the blood or blood plasma concentrations of beta-alanine, L-histidine, and creatine.

During sustained intensive exercise or exercise sustained under conditions of local hypoxia, the accumulation of hydronium ions formed during glycolysis and the accumulation of lactate due to anaerobic metabolism can severely reduce the intracellular pH. The reduced pH can compromise the function of the creatine-phosphorylcreatine system. The decline in intracellular pH also can affect other functions within the cells such as the function of the contractile proteins in muscle fibers. Administering beta-alanine to individual results in elevated muscle beta-alanylhistidine peptide levels and increased total work capacity of the muscle. In addition, chronic dietary supplementation with beta-alanine can increase muscle beta-alanylhistidine peptide concentration and, thus, increase intramuscular buffering capacity.

Beta-alanine supplementation can increase athletic performance by reducing fatigue or reducing the time to the onset of fatigue associated with lactic acid and/or hydrogen ion accumulation during intense exercise or prolonged athletic activity. Typically studies have used beta-alanine supplementation strategies of daily doses of between 200 mg to 6,400 mg, generally following a regime that includes administration of multiple doses of 200 mg, 400 mg or 800 mg tablets multiple times per day, administered at regular intervals for up to eight hours, over periods ranging from 4 to 10 weeks (e.g., see Culbertson et al., Nutrients 2: 75-98 (2010)). After a 10 week supplementation period, the reported increase in intramuscular carnosine content was between 20-80%.

In the methods provided herein, beta-alanine can prevent the progression of and even partially reverse symptoms of sarcopenia and/or frailty. Thus, provided are methods of maintaining or increasing muscle mass and/or strength and/or muscle functionality to treat sarcopenia or frailty in a subject, comprising administering to the subject a therapeutically effective amount of beta-alanine.

Muscle functionality is an important attribute in the ageing subject. Muscle functionality can be a deciding criteria between deciding whether a subject can be considered sufficiently functional to live alone or if the subject requires assistance with everyday tasks. Functional activities such as standing from a seated position, reaching for and retrieving an object, bending, transferring, walking and standing require muscle functionality. These activities also can be influenced by a subject's ability to control their balance. Subjects that exhibit a declines in muscle function also exhibit a loss of balance. Loss of balance and muscle function can lead to increased risk of falls and increased occurrence of falls.

One of the measures used in the art to assess balance is the Berg Balance Test. The Berg Balance Test is an observational performance-based assessment used to evaluate standing balance during a number of functional activities. Generally, the Berg Balance Test includes 14 subtests, which include changing position from sitting to standing, standing without support, sitting without support, changing position from standing to sitting, pivot transfers to go from one chair to another, standing with eyes closed, standing with feet together, reaching forward with an outstretched arm, retrieving an object from the floor, turning to look behind over left and right shoulders while standing, turning 360 degrees (completely turn around in a full circle), placing alternate foot on step or stool while standing unsupported, standing unsupported with one foot directly in front of the other and standing on one leg. The subject is scored on his/her performance during different tasks. The Berg Balance Test has been used in the art to predict falls in elderly persons (e.g., see Berg, Physiotherapy Canada 41(6): 240-245 (1989); Bogle Thorbahn et al, Physical Therapy 76(6): 576-585 (1996) and Perell et al, The Journals of Gerontology: Series A 56(12): M761- M766 (2001)). The test often is administered to subjects who exhibit a decline in function, report a loss of balance, or experience falls. The Berg Balance Test was developed in the early 1990s to measure balance in the elderly. Over the years it has been shown to be a reliable test for evaluating balance. The measured elements of the test are representative of daily activities that require balance, such as sitting, standing, leaning over to pick up an object and stepping. Some tasks are rated according to the quality of the performance of the task, while others are evaluated by the time required to complete the task. The scores assigned for the performance of the task are between 0 (cannot perform the task) to 4 (normal performance of the task). Score below 14 indicated greatly impaired balance while a perfect score of 56 indicates excellent balance (see, e.g., Wood-Dauphinee et ah, Canadian J Rehabilitation 10: 35-50 (1997); Berg et ah, Scand J Rehab Med 27: 27-36 (1995); Berg et ah, Arch Phys Med Rehabilitation 73 : 1073-1083 (1992); and Berg et ah, Physiotherapy Canada 41 : 304-311 (1989)). These values can be used for assessing independent living ability. For example, subjects with a score of less than 20 generally have little to no balance and require a wheelchair or other mechanical means to get from one place to another. Such subjects generally would be unable to live independently without some supervision and/or monitoring. Subjects with a score between 21 and 40 exhibit some loss of balance and muscle function, and generally require some assistance to perform every day activities, such as walking, retrieving an item, such as from a shelf or from the floor, and walking. Subjects with a score or 41 and above exhibit good balance and can perform routine daily tasks with little or no assistance and can live independently. More often than not, subjects with a score of 40 or less tend to experience falls with much higher frequency than subjects with scores higher than 40.

In the methods provided herein, administration of beta-alanine can improve the score of a subject on the Berg balance test. In some embodiments, the score can be improved by 2 or more. In some embodiments, the score can be improved so that it is in the range of 21 to 40. In some embodiments, the score can improved to be in a range between 25 and 45. In some embodiments, the score can improved to be in a range between 30 and 50. In some embodiments, the score can improved to be greater than 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or greater. In some embodiments, administration of beta-alanine to an elderly subject can result in improvement of one or more measures of muscle functionality, including balance, timed up and go test results, functional reach test results, step tests results, rapid step test results, four square step test results, multi-directional reach test results, lateral reach test results, increased ability to stand with the feet together in a side-by-side configuration and/or in a semi-tandem and/or tandem position, and timed stand tests.

Other assessment tests of muscle function in the elderly can be used. These include functional reach tests, lateral reach tests, step tests, four square step test, elderly mobility scale tests, sensory oriented mobility assessment instrument (SOMAI) testing, Fullerton advanced balance scale, Tinetti performance orientated mobility assessment, change of direction while stepping, and hierarchical assessment of balance and mobility (e.g., see Lord et ah, J Am Geriatrics Soc 49(5): 508-515 (2001); Farrell et ah, Topics Geriatric Rehabilitation 20(1): 14-20 (2004); Bennie et al, J Physical Therapy Science 15(2): 93-97 (2003); Dite et al, Archives of Physical Medicine and Rehabilitation 83 : 1566-1571 (2002); Steffen et al., Physical Therapy 82(2): 128-137 (2002); and Langley et al., The Internet Journal of Allied Health Sciences and Practice, Volume 5, Number 4 (2007)).

Beta-alanine can be used as a frailty suppressive agent and as a muscle enhancing agent or muscle increasing agent (providing effects of reducing muscle fatigue in the elderly, preventing elderly hospitalized subjects or patients from being bedridden, shortening rehabilitation period, and improving muscle function) and as an agent for the prophylaxis or treatment of muscle decrease due to disease or misuse, alone or as an adjunct to exercise, and in particular resistance exercise.

In some embodiments, the methods include administering beta-alanine before, concurrent with, or after other optional components such as other active ingredients. In some embodiments, the method includes as a step co-administering a beta-alanine with one or more of the following ingredients: creatine (including its salts (e.g., creatine monohydrate), esters (e.g., creatine ethyl ester), chelates, amides, ethers and derivatives thereof), histidine, vitamin D, Vitamin C, Vitamin B l, Vitamin B2, Vitamin B3, Vitamin B5, Vitamin B6, Vitamin B12, and/or Vitamin K, a mineral, such as chromium, iron, magnesium, sodium, potassium, vanadium, an amino acid, such as L-arginine, L-ornithine, L-glutamine, L-tyrosine, L-taurine, L-leucine, L-isoleucine, L-theanine and/or L-valine and derivatives thereof, one or more peptides, such as L-carnitine, camosine, anserine, balenine, homocarnosine, kyotorphin, and/or glutathione and derivatives thereof, a methylxanthine, such as caffeine, aminophylline or theophylline, antioxidants, such as lutein, zeaxanthine, a flavanol, such as a flavanol extracted from tea or chocolate, and adenosine triphosphates.

Also provided are methods for supplementing the diet of an elderly individual for enhancing the individual's muscle mass and/or muscle size and/or muscle strength and/or endurance and/or muscle functionality. Accordingly, provided are methods of supplementing the dietary intake of an elderly individual comprising administering to the individual an effective amount of a beta-alanine to increase muscle performance or muscle function in the individual. Also provided are methods of improving muscle performance and/or muscle function in an elderly individual, the methods including administering an effective amount of a beta-alanine, alone or in combination with other agents, to the individual.

In any of the methods provided herein, the beta-alanine can be provided as free beta- alanine, or a salt, ester, ether, amide, azide, oxide, or chelate of beta-alanine, including creatine-beta-alaninate salts, or a biological source of beta-alanine. A compound that is a biological source of beta-alanine is a compound that, when administered to the body by any route (for example parenterally, orally, topically), is converted, e.g., via ionic dissolution to constituent ions or by one or more chemical- or enzyme-catalyzed reaction steps, to beta- alanine, which then appears in blood, plasma or serum and is available for uptake into muscle and other tissues.

In the methods provided herein, the free beta-alanine or a biological source thereof may be derivatized as the corresponding salts, esters, enol ethers or esters, acids, bases, solvates, hydrates or pro-drugs prior to administration, as is known in the art.

The effective amount of beta-alanine to be administered can vary according to factors such as age, sex, and weight of the individual. The dosage schedule and regime can be adjusted to provide the optimum response in the individual. Several divided doses can be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of an individual's situation. As will be readily appreciated, the beta-alanine or a composition containing beta-alanine can be administered as a dietary supplement, and can be administered in a single serving or in multiple servings spaced throughout the day. As will be understood by those skilled in the art, servings need not be limited to daily administration, and may be on an every second or third day or other convenient effective basis. The administration on a given day can be in a single serving or in multiple servings spaced throughout the day depending on the exigencies of the situation. The composition can be formulated for bolus administration or for sustained release administration.

Bolus administration of beta-alanine can result in symptoms of paraesthesia. These symptoms include tingling sensations in various parts of the body, particularly in the head and neck region. Once the body is accustomed to the beta-alanine, the tingling symptoms usually stop or become much less pronounced or perceivable. It generally is recommended that a dosage of 1 gram per 2 hour period not be exceeded in order to minimize symptoms of paraesthesia (Derave et ah, Sports Med. 40(3): 247-263 (2010)). Controlled or sustained release formulations have been shown to minimize or eliminate symptoms of paresthesia (e.g., see U.S. Pat. App. Pub. No. US20090220575). Thus, in some embodiments, the methods include administration of beta-alanine in a controlled release formulation. In some methods, the beta-alanine is encapsulated or embedded in a matrix that allows for controlled or sustained release of the beta-alanine from the matrix. A non-limiting example of a sustained-release system is a semipermeable matrix of solid hydrophobic and/or hydophilic polymers. In certain embodiments, sustained release systems can, depending on their chemical nature, release compounds over a period of hours, days, weeks or months.

Controlled release delivery systems are known to those of ordinary skill in the art. They can include polymer based systems, which can include such as poly(lactide-glycolide), copolyoxalates, polycaprolactones, lipids, polyesterimide, polyorthoesters,

polyhydroxybutyric acid, and polyanhydrides and combinations thereof. The formulations also can include en encapsulating coating. Microcapsules of the foregoing polymers containing drugs are described in, for example; U.S. Pat. No. 5,075, 109. Delivery systems also can include lipids, triglycerides, waxes, cholesterol, cholesterol esters and fatty acids or neutral fats such as mono- di- and tri-glycerides, hydrogel release systems, and peptide based systems. Specific examples include, but are not limited to: (a) erosional systems in which the platelet reducing agent is contained in a form within a matrix such as those described in U.S. Pat. Nos. 4,452,775, 4,675,189 and 5,736,152 and (b) diffusional systems in which an active component permeates at a controlled rate from a polymer such as described in U.S. Pat. Nos. 3,854,480, 5, 133,974 and 5,407,686.

The oral ingestion of encapsulated beta-alanine or of a composition including a matrix containing beta-alanine yields a peak concentration (C_max) less than equimolar amounts of orally ingested bolus beta-alanine but greater than that of orally ingested peptides containing beta-alanine, such as anserine or carnosine. In some methods, the controlled release composition is formulated such that the time to peak concentration (T_mx) of the controlled release beta-alanine is longer than that of equimolar amounts of orally ingested bolus beta- alanine but less than that of orally ingested peptides containing beta-alanine, such as anserine or carnosine.

In some embodiments the methods provided herein increase the skeletal muscle level of carnosine in an animal. In some embodiments the methods provided herein increase muscle buffering capacity and thereby allows longer periods of exercise before onset of muscle fatigue in an individual.

SARCOPENIA

Sarcopenia is recognized to be a syndrome characterized by progressive and generalized loss of skeletal muscle mass and strength with a risk of adverse outcomes such as physical disability, poor quality of life and death (see Cruz-Jentoft et ah, Age and Ageing 39(4): 412-423 (2010)). Protocols for assessing the symptoms of sarcopenia in a subject are known in the art (e.g., see Working Group on Functional Outcome Measures for Clinical Trials, J Gereontology A Biol Sci Med Sci 64A(4): 487-491 (2009); and Rolland et al, Journal of Nutrition, Health & Aging 12(7): 433-450 (2008)). The European Working Group on Sarcopenia in Older People recommends using the presence of both low muscle mass and low muscle function (strength or performance) for the diagnosis of sarcopenia. It has been suggested that the term "dynapenia" be used to describe age-associated loss of muscle strength and function but those in the art have embraced "sarcopenia" and continue to use that term. The methods provided herein also treat or prevent the symptoms of dynapenia.

Sarcopenia can be defined using a number of criteria, including the amount of muscle and its function. The quantifiable variables are muscle mass, strength and physical performance. The challenge has been how to accurately quantify each of these attributes.

Muscle mass can be measured using any method known in the art (e.g., see U.S. Pat. No. 5,628,328; Heymsfield et al, Am J Clin Nutrition, 37: 478-494 (1983); Visser et al, J Appl Physiol 87: 1513-1520 (1999); Ohkawa ei a/., Am J Clin Nutrition, 71(2): 485-490 (2000)). Muscle mass can be quantified by using, e.g., an imagining technique, such as computed tomography (CT scan), dual energy X-ray absorptiometry (DXA) and magnetic resonance imaging (MRI). Because of the ability of CT and MRI to separate images of fat from other soft tissues of the body, these scanning methods are considered the ultimate standards for estimating muscle mass in research (see Cruz-Jentoft et al, Age and Ageing 39(4): 412-423 (2010)). Both CT scans and MRI involve procedures that are relatively expensive, thereby hampering their widespread use. Bioimpedance analysis also can be used to estimate lean body mass and fat volume of an individual. Bioimpedance analysis measurement techniques are well known in the art (e.g., see U.S. Pat. Nos. 5,722,396;

7,499,745; Nunez et al, J Parenteral and Enteral Nutrition 23 : 96-103 (1999); and Pietrobelli et al, European Journal of Clinical Nutrition 58: 1479-1484 (2004)). It has been reported in the art that results from bioimpedance analysis under standard conditions correlate well with MRI predictions (e.g., see Janssen et al, J Appl Physiol 89: 465-471 (2000)) and it has been shown that bioimpedance analysis measurement correlates well to muscle function (e.g., see Norman et al, Clin Nutr 28: 78-82 (2009)).

In some methods, an amount of beta-alanine is provided to increase the carnosine content in an elderly subject. Muscle carnosine content can be assessed using any method known in the art. For example, muscle carnosine content can be assessed in vivo by ^XH-MRS (magnetic resonance spectroscopy) using a whole body 3.0T MRI scanner (Achieva Intera, Philips, Best, The Netherlands) and a 14 cm diameter ^-surface coil to detect labelled carnosine (such as from synthesis using labeled beta-alanine). In an exemplary method, the surface coil can be placed centered under the calf muscle of one leg. The scanner body coil can be used to obtain conventional anatomical Tl-weighted magnetic resonance images in the three orthogonal planes. Spectrum raw data can be analyzed using any method or software known in the art, such as by Java Magnetic Resonance User Interface software (e.g., see Naressi et al., Computers in Biology and Medicine 31(4): 269-286 (2001); and Liao, Computer Methods and Programs in Biomedicine 67(2): 155-162 (2002)). The analysis can include apodization to 5Hz, Fourier transform, phase correction or any combination thereof. The carnosine signal in the muscle can be quantified relative to an external reference.

One measure of sarcopenia uses the appendicular skeletal muscle mass (kg/height²

(m²)), where sarcopenia is defined as being less than two standard deviations below the mean of a young reference group (i.e., the t-score). A t-score is determined by measuring the axial skeletal muscle mass of a subject, typically by DXA (i.e., dual energy x-ray absorptiometry) or a similar and reproducible measure. The measurement of axial skeletal muscle mass can be used to follow the progress of the subject to determine if treatment is slowing, preventing, or reversing muscle mass decline. Examples of t-scores include 3, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3,

2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0,—0.1,—0.2,—0.3,—0.4,—0.5,—0.6,—0.7,—0.8,—0.9,—1.0,—1.1,—1.2,—

1.3,—1.4,—1.5,—1.6,—1.7,—1.8, 15 —1.9,—2.0,—2.1,—2.2,—2.3,—2.4,—2.5, —2.6,—2.7,—2.8,—2.9,—3.0,—3.1,—3.2,—3.3,—3.4,—3.5,—3.6,—3.7,—3.8,—

3.9,—4.0,—4.1,—4.2,—4.3,—4.4,—4.5,—4.6,—4.7,—4.8,—4.9,—5.0,—5.1,—5.2, — 5.3,— 5.4,— 5.5,— 5.6,— 5.7,— 5.8,— 5.9, and— 6.0. Typically subjects with negative t- scores are more likely to be exhibiting symptoms of sarcopenia and thus are more often treated for sarcopenia. A patent that is at risk of losing muscle 20 function and/or mass or who has a medical need to maintain muscle function and/or mass also can be a subject for treatment in accordance with the present methods even if their t-score is 0 or greater.

Another parameter that can be measured for the diagnosis of and confirmation of efficacy of interventions for the treatment of sarcopenia or frailty is muscle strength. The generally accepted measurement criterion for the maximum tension which can be exerted by a muscle is the maximum amount of force a muscle can exert on a body part. In physiology, this is referred to as the maximum strength of the muscle and might be expressed, for example, in kilograms per square centimeter of muscular section. Muscle strength can be expressed in kilograms, Newtons, pounds or inch-pounds and Newton-meters. Techniques to measure muscle strength are known in the art (e.g., see U.S. Pat. Nos. 3, 133,355; 6,063,044; and 6,706,003 and U.S. Pat. Appl. Pubs. US20050245848 and US20010029342). These techniques include isometric muscle strength testing methods, isometric manual muscle testing, and comparison of force and displacement measurements.

Muscle strength measurements can include muscle strength of lower limbs, of upper limbs, or their combination. In some instances, the measurement of upper limbs is preferable to the measurement of lower limbs, while in other circumstances the reverse is true. For example, in assessing load bearing strength, measurement of the upper limbs alone may be a good measurement of muscle strength. In the case of the ability to relocate massive objects, the measurement of the combination of upper and lower limb strength may be indicated. For evaluating gait, walking and stair climbing, measurement of lower limb strength may be indicated.

One muscle strength measurement used in the art is isometric hand grip strength (e.g., see U.S. Pat. Nos. 4,674,330; 5, 170,663; and 6,678,549). It has been shown that hand grip strength correlates well with other muscle strength testing techniques, including knee extension torque, lower extremity muscle power and with calculations of muscle strength based on cross-sectional area of calf muscle (e.g., see Laurentani et al., J Appl Physiol 95: 1851-1860 (2003)). Handgrip strength has been demonstrated as a predictor of muscle function, including in the oldest old (e.g., see Taekema et al., Age Ageing 39(3): 331-337 (2010)). Grip strength can be measured by any method known in the art, including by use of a handheld dynamometer in comparison to a reference population (Febrer et al., J Rehabil Med. 42(3): 228-231 (2010)).

Other methods of muscle strength can be used. For example, lower extremity muscle strength can be measured using a leg extension movement, such as by using the method described by Bassey et al. (Eur J Appl Physiol 60: 385-390 (1990)). These measurements can be performed isokinetically or isometrically. The isokinetic strength tests, particularly of the knee and ankle, have been demonstrated to be reproducible in older adults (e.g., see

Hartmann et al, Gerontology 55: 259-268 (2009)).

The Working Group on Functional Outcome Measures for Clinical Trials has described a number of physical performance test procedures for testing the elderly for symptoms or sarcopenia and/or frailty (J Gerontol A Biol Sci Med Sci 63 : 160-164 (2008)). These include the stair climb power test, 6-min walk test and usual gait speed. Other physical performance tests for assessing lower extremity function are described in the art (e.g., see Guralnik et ah, J Gerontol 49:M85-M94 (1994)). After years of testing, it has been suggested that gait speed can be used as a predictor of adverse outcomes (Abellan van Kan et ah, J Nutr Health Aging 13 : 881-889 (2009)).

Another protocol useful for assessing muscle functionality is the timed get-up- and-go test (e.g., see U.S. Pat. No. 6,972, 124). The timed get-up-and-go test is a gross measure of balance and lower extremity function (e.g., see Mathias et ah, Arch Phys Med Rehabil 67: 387 (1986); Morris et ah, Phys Ther 81 : 810 (2001); Okumiya J Am Geriatr Soc 46: 928 (1998); and Shumway-Cook Phy Ther 80: 896 (2000)). In some protocols, the time needed by a subject to rise from a standard arm chair, walk to a line on the floor three meters away, turn, return, and sit down again is measured.

Another testing procedure for measuring muscle functionality is the stair climb muscle power test (e.g., see U.S. Pat. No. 6,972,124). It is known in the art that the stair climb test is a clinically relevant measure of leg power impairments in at-risk older adults and can be used to assess mobility performance (e.g., see Roig et ah, Am. J. Respir. Crit. Care Med. 181 : A3583 (2010); Bean et ah, Arch Phys Med Rehabil 88: 604-609 (2007) and Herman et ah, J Gerontol A Biol Sci Med Sci. 60(4): 476-480 (2005)).

Another testing procedure for assessing muscle performance is the timed stand test. The timed stand test is a measure of leg muscle strength and assesses the time needed for a subject to rise from a chain a given number of times as quickly as possible (e.g., see Csuka et ah, Am J Med 78: 77-81 (1985). The test is a simple method for measuring lower extremity muscle strength. In some protocols, the test measures the time needed for a subject to stand 10 times from a standard chair.

Other testing procedures can be used to assess muscle function in an elderly subject. These include up and down tests, functional reach tests, lateral reach tests, step tests, four square step test, elderly mobility scale tests, sensory oriented mobility assessment instrument (SOMAI) testing, Fullerton advanced balance scale, Tinetti performance orientated mobility assessment, change of direction while stepping, and hierarchical assessment of balance and mobility (e.g., see Lord et ah, J Am Geriatrics Soc 49(5): 508-515 (2001); Farrell et ah, Topics Geriatric Rehabilitation 20(1): 14-20 (2004); Bennie et ah, J Physical Therapy Science 15(2): 93-97 (2003); Dite et ah, Archives of Physical Medicine and Rehabilitation 83: 1566-1571 (2002); and Langley et ah, The Internet Journal of Allied Health Sciences and Practice, Volume 5, Number 4 (2007)). In the methods provided herein, the beta-alanine improves muscle functionality and also can improve exercise tolerance. Other parameters that can be measured to assess efficacy of an intervention for the treatment or prevention of sarcopenia and/or frailty includes results from physical capacity tests. These tests include specific objective physiological measurements such as maximum oxygen uptake (V0_2max) (see MacVicar et ah, Nurs Res 3 : 348-351. (1989)) and ventilatory anaerobic threshold (VAT) (see Kreider et ah, Med Sci Sports Exerc. 22(2): 250-256 (1990). Anaerobic exercise testing also can be performed to assess physical capacity (e.g., see U.S. Pat. Nos. 6, 176,241). An exemplary test is a cycle ergometer test to the limit of tolerance (e.g., see Puente-Maestu et al., Respiration 70: 367-370 (2003). In some testing, a subject performs an incremental test on a motorized treadmill to determine the ventilatory anaerobic threshold (VAT) and

A "Wingate test," which is a cycle ergometer test used to measure muscle work over a relatively short period (e.g., 30 seconds) also can be used. In some testing protocols, assessment of anaerobic exercise capacity is evaluated by subjecting a subject to one repetition square-wave transition from rest to exercise intensity corresponding to 75% (Delta), i.e., 75% of the difference between VAT and

to the limit of tolerance (TLIM), which is similar to the cycle ergometer test described by Puente-

also can be determined by having subjects cycle on a stationary cycle ergometer using four 5- min steady-state stages [100, 150, 200, and 250 work rate (W)] followed by a progressive increase in work of 10 W/min until voluntary exhaustion. A work rate of 75%

can be calculated for each subject from the linear function of oxygen uptake at the four steady-state work rates, and maximal oxygen uptake against the work rate of the four steady state work rates and maximal power output.

Sarcopenia generally is observed in older people, generally those over age 60 or 65, but it also can manifest in younger adults. In clinical practice, primary sarcopenia refers to age-related sarcopenia when no other cause is evident but ageing itself. When one or more other causes are evident, the loss of muscle mass and strength or function is considered secondary sarcopenia. Examples of secondary sarcopenia include "lack of activity" causes, such as bed rest, confinement to a wheelchair or sedentary lifestyle, sarcopenia related to improper nutrition, such as inadequate dietary intake of protein, nutrient malabsorption, or gastrointestinal disorders or anorexia, and sarcopenia caused by disease, such as

inflammatory disease, malignancy or cancer, or an endocrine disease.

Treating sarcopenia includes slowing its progression, stopping its progression, and partially reversing its progression. An example of slowing the progression of sarcopenia would be to change the length of time a subject would go from a t-score of— 1.5 to a score of — 2 (e.g., if such a progression would normally take 5 years, then treating as used herein could slow this change to 10 years). Examples of partial reversal include reducing a t-score 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 or more units (e.g., moving from a t-score of— 2 to a t-score of-1.9,—1.8,—1.7,—1.6,—1.5,—1.4,—1.3,—1.2,—1.1, etc.). In some embodiments, the method includes identifying a subject having a t- score selected from among (a) <-3, (b) <-2.5, (c) <-2, (d) <-1.5, (e) <-1.0, and (f) <-0.5; and administering beta- alanine to slow, stop or reverse the progression of sarcopenia.

Treating sarcopenia also includes delaying the onset of sarcopenia. For example, if a typical male age 60 would begin to see signs of sarcopenia by age 65, treatment could delay the onset 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more years. Thus, treating sarcopenia would include treating subjects who have not yet been diagnosed with sarcopenia, but who would be vulnerable or expected to be vulnerable to developing sarcopenia in the future. Subjects who are vulnerable or expected to be vulnerable in the future also include (a) subjects using glucocorticoid steroids, (b) subjects with chronic infections, (c) subjects with chronic inflammatory conditions (e.g., inflammatory bowel disease), (d) subjects with cancer and (e) patents with a family history of sarcopenia. In some embodiments, a further decline in t- score is prevented via treatment with beta-alanine for at least a year. In some embodiments, an increase in the t-score of subject is obtained via treatment with beta-alanine for at least a year.

Another type of subject that would benefit from the methods provided herein is a subject who has suffered some loss of muscle mass, but who does not suffer from a condition that interferes with acts of daily living and/or prevents the subject from living an independent life (e.g., a subject who might soon need assisted living).

The age or age range of the subject can vary depending on their susceptibility to sarcopenia. Examples of ages and age ranges include (a) 40-45, (b) 45-50, (c) 50-55, (d) 55- 60, (e) 60-65, (f) 65-70, (g) 70-75, (h) 75-80, (i) 80-85, (j) 85-90, or older.

FRAILTY

Frailty generally is associated with old age. Frailty is a geriatric syndrome resulting from age-related cumulative declines across multiple physiologic systems, with impaired homeostatic reserve and a reduced capacity of the individual to withstand stress, thus increasing vulnerability to adverse health outcomes including falls, hospitalization, institutionalization and death (see Cruz-Jentoft et ah, Age and Ageing 39(4): 412-423 (2010)). Protocols for assessing the symptoms of frailty in a subject are known in the art (e.g., see Working Group on Functional Outcome Measures for Clinical Trials, J Gereontology A Biol Sci Med Sci 64A(4): 487-491 (2009)). Because of the ageing world population, age- related frailty has emerged as an important public health problem. Frailty can result in impaired mobility and decreased quality of life, and is associated with increased risk of falls (Roubenof, J Nutr. 129: 256S-259S (1999)). Evidence for a phenotype of frailty has been proposed, based on readily identifiable physical aspects, which includes the manifestation of three or more of the following characteristics: unintended weight loss, exhaustion, weakness, slow gait speed and low physical activity (Fried et ah, J Gerontol A Biol Sci Med Sci 56:M146-56 (2001)).

Several studies have shown that lean body mass decreases progressively after the mid-

20s and that age-related losses in fat-free mass, appendicular muscle mass, and total muscle mass are all linear in both men and women (Baumgartner et ah, Am J Epidemiol. 147: 755- 763 (1998) and Melton et ah, J Am Geriatr Soc. 48(6):625-630 (2000)).

Frailty and sarcopenia include similar underlying etiologies and thus overlap to some extent, but frailty also includes other attributes, such as changes in cognitive status (Bauer et ah, Exp Gerontol 43 :674-678 (2008)).

CHANGE IN MUSCLE IN SUBJECTS WITH SARCOPENIA OR FRAILTY

Skeletal muscle fibers are generally classified as type I (oxidative/slow) or type II (glycolytic/fast) fibers. They display marked differences in respect to concentration, metabolism, and susceptibility to fatigue. Type I fibers are mitochondria-rich and mainly use oxidative metabolism for energy production, which provides a stable and long-lasting supply of ATP, and thus are fatigue-resistant. Type II fibers comprise three sub-types: Ila, IIx, and lib. Type lb fibers have the lowest levels of mitochondrial content and oxidative enzymes, rely on glycolytic metabolism as major energy source, and are susceptible to fatigue, while the oxidative and contraction functions of type Ila and IIx lie between type I and lib. In adults, skeletal muscle can modulate between different fiber types, such as in response to exercise training (e.g., see Wang et ah, PLoS Biology 2(10): e294 (2004)).

In older adults, extended hospitalization can result in disuse atrophy leading to a potential loss of the ability for independent living and to a cascade of physical decline.

Moreover, the physical aging process profoundly affects body composition, including significant reductions in lean body mass and increases in central adiposity. In addition, it is possible that the age-associated decrease in muscle mass, and subsequently in muscle strength and endurance, can be a critical determinant for functional loss, dependence and disability. Muscle weakness also is a major factor predisposing the elderly to falls and the resulting morbidity and mortality.

Muscle function can become compromised by many mechanisms. Examples include the frailty associated with old age and sarcopenia. Determination of the muscle fiber composition in athletes revealed that elite endurance athletes have relatively more type I fibers than type II fibers in the trained musculature. Marathoners also tend to have more type I fibers. It was suggested that type I fiber might be a factor governing physical endurance capacity. Ageing and physical inactivity are conditions associated with a decrease in type I fibers. It appears that the muscle oxidative capacity is a crucial factor for determining endurance and fatigue resistance. There seem to be an adaptive metabolic response of skeletal muscle to endurance exercise by controlling the number of oxidative muscle fibers (type I fibers).

Muscle wasting is characterized by a progressive loss of muscle mass, weakening and degeneration of muscles especially the skeletal or voluntary muscles and the cardiac muscles. The processes by which atrophy and hypertrophy occur are conserved across mammalian species. Multiple studies have demonstrated that the same basic molecular, cellular, and physiological processes occur during atrophy in both rodents and humans. Thus, rodent models of skeletal muscle atrophy have been successfully utilized to understand and predict human atrophy responses. As the body ages, an increasing proportion of skeletal muscle is replaced by fibrous tissue. Therefore, normal aging in humans is associated with progressive decrease in skeletal muscle mass and strength and function, which contributes to frailty and falls. Conditions resulting in muscle wasting can arise from disuse conditions such as long term immobilization due to illness or disability such as confinement in a wheelchair, prolonged bed rest, bone fracture or trauma. It is estimated that bed-rest after surgery causes loss of skeletal muscle mass of approximately 10% per week. Untreated muscle wasting disorders can have serious health consequences. The changes that occur during muscle wasting can lead to a weakened physical state resulting in poor performance of the body and detrimental health effects.

Thus, muscle atrophy can seriously limit the rehabilitation of subjects after immobilization. Muscle wasting due to chronic diseases can lead to premature loss of mobility and increase the risk of disease-related morbidity. Muscle wasting due to disuse is an especially serious problem in elderly, who may already suffer from age-related deficits in muscle function and mass, leading to permanent disability and premature death. Despite the clinical importance of the condition, few treatments exist to prevent or reverse the condition.

Muscle wasting includes the progressive loss of muscle mass and/or to the progressive weakening and degeneration of muscles, including the skeletal or voluntary muscles, which control movement, cardiac muscles, which control the heart (cardiomyopathies), and smooth muscles. Chronic muscle wasting is a chronic condition (i.e. persisting over a long period of time) characterized by progressive loss of muscle mass, weakening and degeneration of muscle.

The loss of muscle mass that occurs during muscle wasting can be characterized by muscle protein degradation by catabolism. Protein catabolism occurs because of an unusually high rate of protein degradation, an unusually low rate of protein synthesis, or a combination of both. Muscle protein catabolism, whether caused by a high degree of protein degradation or a low degree of protein synthesis, leads to a decrease in muscle mass and to muscle wasting.

Health status during the aging process is significantly influenced by nutrition.

Malnutrition has been shown to increase mortality in the aged and protein energy

undernourishment has been shown to accelerate the age associated rate of sarcopenia and frailty. With inadequate nutritional intake during aging, a shift toward an increased amount of body fat mass takes place and the ratio of muscle mass to fat mass decreases.

The physiological mechanism for the decline in muscle mass is unknown. Growth hormone secretion declines progressively from mid puberty, and growth hormone is known to increase muscle mass. Subjects with growth hormone deficiency have reduced muscle mass and increased fat mass. Growth hormone replacement increases the muscle mass and leads to a reduction in fat mass.

Maintenance of muscle mass depends on two different processes: maintaining function and exercise that allow muscle mass to be maintained or built; and, an ability to maintain muscle mass that depends on nutrition, neural input, and hormonal state. The peak muscle mass is observed at the time of mid-puberty and muscle mass progressively declines and is detectably reduced by the age of 45 years and continues to progressively decline. This decline in muscle mass appears to be dependent on growth hormone secretion, which declines with age. U.S. Pat. No. 6, 194,402 (Bach et al.) describes the use of growth hormone secretagogues, including MK-677 for treating long-term muscle loss or wasting. Measures of physical performance, particularly muscle function and strength, are objective tests of a subject's performance of standardized tasks, and can be evaluated according to predetermined criteria that can include counting repetitions or timed activity. A decline in physical performance results in increased odds that the subject can suffer an adverse event such as an injurious fall and/or fracture. A decline in physical performance also can result in the subject having to be admitted to a nursing home and/or developing functional dependence in activities of daily living.

Sarcopenia has been identified as a contributor to loss in muscle functionality.

Among the daily activities impaired by sarcopenia are rising from a chair, climbing stairs and even walking. Muscle functionality can be a deciding criteria between deciding whether a subject can be considered sufficiently functional to live alone or if the subject requires an amount of assistance with everyday tasks that would preclude the subject from living alone. Functional activities such as standing from a seated position, reaching for and retrieving an object, bending, transferring, walking and standing require muscle functionality. Thus, although loss of muscle mass, particularly the loss of type II muscle fibers can result in diminished strength and power-generating capacity (Harris, J utr. 127: 1004S-1006S (1997)), loss of muscle function can more directly impact the quality of life of an elderly subject. A particular effect of loss of muscle function is diminished balance in the elderly. An apparent result of this loss of balance is the frequent falls experienced by the elderly.

Therefore, interventions that increase muscle function can improve balance and reduce the risk of falling.

Administration of beta-alanine to an elderly subject can improve muscle function. Muscle function can be assessed by any technique known in the art. These include the Berg Balance Test, functional reach tests, lateral reach tests, step tests, four square step test, elderly mobility scale tests, sensory oriented mobility assessment instrument (SOMAI) testing, Fullerton advanced balance scale, Tinetti performance orientated mobility assessment, change of direction while stepping tests, timed up and go tests, timed stand tests, clinical test of sensory interaction and balance, and hierarchical assessment of balance and mobility (e.g., see Berg, Physiotherapy Canada 41(6): 240^~245 (1989); Bogle Thorbahn et al, Physical Therapy 76(6): 576-585 (1996) and Perell et al, The Journals of Gerontology: Series A 56(12): M761-M766 (2001)). (e.g., see Lord et al, J Am Geriatrics Soc 49(5): 508-515 (2001); Farrell et al, Topics Geriatric Rehabilitation 20(1): 14-20 (2004); Bennie et al, J Physical Therapy Science 15(2): 93-97 (2003); Dite et al, Archives of Physical Medicine and Rehabilitation 83: 1566-1571 (2002); and Langley et ai, The Internet Journal of Allied Health Sciences and Practice, Volume 5, Number 4 (2007)).

NUTRITION AND SARCOPENIA AND/OR FRAILTY

Loss of skeletal muscle mass occurs with ageing. For example, in the 60 years after a person's 21^st birthday, the decline in skeletal muscle mass can be as high as 40% (Evans, J Gerontol. 50A: 147-150 (1995); Schoeller, Am J Clin Nutr. 50: 1 176-1 181 (1989)). The depletion of muscle mass does not necessarily result in weight loss in the subject because there can be a corresponding accumulation of body fat, which could mask the loss of muscle mass. The age-associated changes in muscle composition result from a combination of different factors, including a general decline in muscle protein turnover (Nair, J Gerontol

(Biol Med Sci). 50A: 107-112 (1995)). Some loss of muscle mass can be attributed to dietary decline of protein intake, since it has been reported that protein intake appears to decline in the elderly (Walrand et al, Curr Opin Clin Nutr Metab Care 8: 89-94 (2005)). One of the intervention strategies suggested in the art to reduce the progression of sarcopenia includes the provision of supplemental doses of protein and/or targeted control of daily protein consumption to slow or prevent muscle protein catabolism (e.g., see Paddon- Jones et al., Am J Clin Nutrition 87(5): 1562S-1566S (2008)). It has been suggested that increasing daily protein intake to 0.8 g/kg body weight per day may be able to improve muscle protein anabolism and reduce loss of muscle mass in the elderly (e.g., see Paddon- Jones et al., Am J Clin Nutrition 87(5): 1562S-1566S (2008)). However, there is no consensus as to what would constitute an optimal value of protein in the daily diet of an elderly subject, nor what proteins or combination of amino acids are optimal for reversing muscle loss with ageing.

In the methods provided herein, beta-alanine can be administered to an elderly subject in combination with a dietary protein or a protein supplement comprising a combination of amino acids. In some methods, the protein is a whey protein, or a whey protein isolate, a soy protein or soy protein isolate, a casein or any combination thereof. In some methods, the protein is fortified with one or more essential amino acids. In some methods, beta-alanine is administered with one or more essential amino acids, particularly leucine. In some methods, the beta-alanine can be administered with a combination of isolated whey proteins and isolated casein proteins. In some methods, the beta-alanine is co-administered with a dietary source of protein high in essential amino acids, such as beef or chicken. In some methods, the beta-alanine is co- administered with a dietary source of protein high in omega-3 fatty acids. In some methods, the beta-alanine is co-administered with a dietary source of protein and a source of omega-3 fatty acids.

EXERCISE AND SARCOPENIA AND/OR FRAILTY

It is suggested in the art that increasing physical activity has a positive effect on skeletal muscle mass and that the decrease in activity with advancing age may contribute to the onset of sarcopenia (Roubenoff et al, Nutr Rev. 51 : 1-11 (1993)). The intensity of strength training highly influences the magnitude of improvement in muscle strength, particularly in the elderly. If the intensity of strength training is low, only low to moderate increases in muscle strength are observed, while high-intensity, resistance training programs in healthy older men resulted in significantly high increases in muscle strength (Frontera et al, J Appl Physiol. 64: 1038-1044 (1988) and Aniansson et al, Clin Physiol. 1 :87-98 (1981)).

Resistance training has been shown to be particularly successful in increasing muscle mass and strength, including in the elderly with a mean age of 90 exhibiting symptoms of frailty (Fiatarone et al, N Engl J Med. 330: 1769-1775 (1994)). It has been found that administering beta-alanine to elderly subjects allows for prolonged or increased exercise, including resistance training, by minimizing muscle fatigue as compared to exercising without beta-alanine supplementation. In the methods provided herein, the beta-alanine also can improve exercise tolerance.

PROVIDING BETA-ALANINE IN COMBINATION WITH ANOTHER AGENT

In the methods provided herein, the beta-alanine can be co-administered with, and the pharmaceutical compositions can include, other medicinal agents, pharmaceutical agents and/or adjuvants. Suitable medicinal and pharmaceutical agents include modulators of one or more of skeletal myosin, skeletal actin, skeletal tropomyosin, skeletal troponin C, skeletal troponin T, skeletal troponin T, and skeletal muscle, including fragments and isoforms thereof, and the skeletal sarcomere and other suitable therapeutic agents such as anti-obesity agents.

Exemplary medicinal and pharmaceutical agents that can be co-administered with beta-alanine include, for example: orlistat, sibramine, diethylpropion, phentermine, benzaphetamine, phendimetrazine, estrogen, estradiol, levonorgestrel, norethindrone acetate, estradiol valerate, ethinyl estradiol, norgestimate, conjugated estrogens, esterified estrogens, medroxyprogesterone acetate, testosterone, insulin-derived growth factor, human growth hormone, riluzole, cannabidiol, prednisone, albuterol and non- steroidal anti-inflammatory drugs. Additional exemplary medicinal and pharmaceutical agents that can be coadministered with beta-alanine include TRH, diethylstilbesterol, theophylline, enkephalins, E series prostaglandins, sulbenox, growth hormone secretagogues, such as GHRP-6, GHRP-1 (disclosed in U.S. Pat. No. 4,411,890 and publications WO 89/07110 and WO 89/0711 1), GHRP-2 (disclosed in WO 93/0408 1), NN703 (Novo Nordisk), LY444711 (Lilly), MK-677 (Merck), CP424391 (Pfizer) and B-HT920, growth hormone releasing factor and its analogs, growth hormone and its analogs and somatomedins including IGF-1 and IGF -2, alpha- adrenergic agonists, such as clonidine or serotonin 5-HTD agonists, such as sumatriptan, agents which inhibit somatostatin or its release, such as physostigmine, pyridostigmine, parathyroid hormone, PTH(l-34), and bisphosphonates, such as alendronate, estrogen, testosterone, selective estrogen receptor modulators, such as tamoxifen or raloxifene, progesterone receptor agonists ("PRA"), such as levonorgestrel, medroxyprogesterone acetate (MP A), aP2 inhibitors, such as those disclosed in U.S. Ser. No. 09/519,079 filed Mar. 6, 2000, PPAR gamma antagonists, PPAR delta agonists, beta 3 adrenergic agonists, such as AJ9677 (Takeda/Dainippon), L750355 (Merck), and CP331648 (Pfizer), other beta 3 agonists, such as those disclosed in U.S. Pat. Nos. 5,541,204, 5,770,615, 5,491, 134,

5,776,983 and 5,488,064, a lipase inhibitor, such as orlistat or ATL-962 (Alizyme), a serotonin (and dopamine) reuptake inhibitor, such as sibutramine, topiramate or axokine, anorectic agents, such as dexamphetamine, phentermine, phenylpropanolamine or mazindol, an HIV or AIDS therapy, such as indinavir sulfate, saquinavir, saquinavir mesylate, ritonavir, lamivudine, zidovudine, lamivudine/zidovudine combinations, zalcitabine, didanosine, stavudine, and megestrol acetate, an anti-resorptive agent, vitamin D, vitamin D analogues, and cathepsin K inhibitors.

In some embodiments, the methods include co-administration of beta-alanine and vitamin D or a vitamin D analog. In some embodiments, the methods include providing at least 600 IU (15 μg) vitamin D per day. In some embodiments, the methods include providing at least 1,000 IU vitamin D per day. In some embodiments, the methods include providing at least 1,500 IU vitamin D per day. In some embodiments, the methods include providing at least 2,000 IU vitamin D per day. In some embodiments, the methods include providing at least 2,500 IU vitamin D per day. In some embodiments, the methods include providing at least 5,000 IU vitamin D per day. In some embodiments, the methods include providing at least 7,500 IU vitamin D per day. In some embodiments, the methods include providing at least 10,000 IU vitamin D per day. In some embodiments, the methods include providing at least 15,000 IU vitamin D per day. In some embodiments, the methods include providing at least 20,000 IU vitamin D per day. In some embodiments, the methods including providing between 500 and 25,000 IU vitamin D per day. In some embodiments, the methods include co-administering beta-alanine and a vitamin D or a vitamin D analog, where the amount of vitamin D or a vitamin D analog administered results in a blood serum level of 25- hydroxyvitamin D greater than 50 ng/mL, such as between 60 ng/mL to 60 ng/mL.

In any of the methods provided herein, the beta-alanine also can be co-administered with creatine and/or histine. In some embodiments, the beta-alanine is co-administered with a nutraceutical. In some embodiments, the beta-alanine is co-administered with a vitamin, a mineral, an omega 3 fatty acid or combinations thereof.

DOSAGE FORMS

In the methods provided herein, the beta-alanine can be provided in any dosage form. Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the beta-alanine can be admixed with at least one inert pharmaceutically acceptable carrier such as sucrose, lactose, or starch. Such dosage forms also can include, as is normal practice in the pharmaceutical arts, additional substances and/or inert diluents, e.g., lubricating agents, such as magnesium stearate, buffering agents, such as sodium citrate, magnesium carbonate, magnesium bicarbonate, calcium carbonate, and calcium bicarbonate, and can include additional active ingredients. Dosage forms in the form of tablets and pills can additionally be prepared with enteric coatings. In some embodiments, the dosage form is a liquid, dissolvable film or a chewable form, particularly for individuals that have a difficult time swallowing tablets or capsules. In some embodiments, the dosage form can include flavoring and/or sweetening agents.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Besides such inert diluents, compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring and perfuming agents.

The methods provided herein include administering beta-alanine via parenteral administration. Dosages for such administration can include sterile aqueous or non- aqueous solutions, suspensions, or emulsions. Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. Such dosage forms also can contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. The formulations can be sterilized by, for example, filtration through a bacteria-retaining filter, by

incorporating sterilizing agents into the compositions, by irradiating the compositions, or by heating the compositions. They also can be manufactured in the form of sterile solid compositions, which can be dissolved in sterile water, or some other sterile injectable medium immediately prior to use.

The methods provided herein also include administering beta-alanine in a composition for rectal or vaginal administration, such as suppositories, which may contain, in addition to beta-alanine, excipients such as cocoa butter or a suppository wax. The methods provided herein also include administration of beta-alanine via nasal or sublingual administration, using formulations prepared using standard excipients that are well known in the art.

In some embodiments of the methods herein, supplemental vitamins and/or minerals also can be administered, before, concurrent with, or after administration of the beta-alanine. Suitable minerals can include one or more minerals or mineral sources with a focus on use of critical vitamins or minerals associated with benefit in the aging process. These include vitamin D, calcium, the family of B vitamins, vitamin A, E and C. Non-limiting examples of minerals include, without limitation: chloride, sodium, iron, chromium, copper, iodine, zinc, magnesium, manganese, molybdenum, phosphorus, potassium, and selenium. Suitable forms of any of the foregoing minerals include soluble mineral salts, slightly soluble mineral salts, insoluble mineral salts, chelated minerals, mineral complexes, non-reactive minerals such as carbonyl minerals, and reduced minerals, and combinations thereof.

In some embodiments, the methods provided herein include administering vitamins in conjunction with a beta-alanine. The vitamins can be fat-soluble or water soluble vitamins. Suitable vitamins include but are not limited to vitamin C, vitamin A, vitamin E, vitamin B12, vitamin K, riboflavin, niacin, vitamin D, vitamin B6, folic acid, pyridoxine, thiamine, pantothenic acid, and biotin. The form of the vitamin can include salts of the vitamin, derivatives of the vitamin, compounds having the same or similar activity of a vitamin, and metabolites of a vitamin.

Tablets and pills can be compressed, multiply compressed, multiply layered, and/or coated. The coating cab be single or multiple. In one embodiment, the coating material includes a polysaccharide or a mixture of saccharides and glycoproteins extracted from a plant, fungus, or microbe. Non-limiting examples include corn starch, wheat starch, potato starch, tapioca starch, cellulose, hemicellulose, dextrans, maltodextrin, cyclodextrins, inulins, pectin, mannans, including glucomannans and galactomannans, gum arabic, locust bean gum, mesquite gum, guar gum, gum karaya, gum ghatti, tragacanth gum, carrageenans, ,β-glucans, agar, alginates, chitosans, xanthan gum, rhamsan gum or gellan gum. In some embodiments, the coating material can include a protein. Suitable proteins include, but are not limited to, gelatin, casein, collagen, whey proteins, soy proteins, rice protein, and corn proteins. In some embodiments, the coating material can include a fat or oil, and in particular, a high temperature melting fat or oil. The fat or oil can be hydrogenated or partially hydrogenated, and can be derived from a plant. The fat or oil can include glycerides, free fatty acids, fatty acid esters, or a mixture thereof. In some embodiments, the coating material contains an edible wax. Edible waxes can be derived from animals, insects, or plants. Non-limiting examples include beeswax, lanolin, bayberry wax, carnauba wax, and rice bran wax. Tablets and pills also can be prepared with enteric coatings.

In some embodiments, the methods include administering beta-alanine in a controlled release or sustained release formulation. For example, the beta-alanine can be provided in a controlled release matrix, where the beta-alanine is dispersed within a matrix that can be either insoluble, soluble, or a combination thereof. Controlled release matrix dosage forms of the insoluble type are also referred to as "insoluble polymer matrices", "swellable matrices", or "lipid matrices" depending on the components that make up the matrix. Controlled release matrix dosage forms of the soluble type also are referred to as "hydrophilic colloid matrices", "erodible matrices", or "reservoir systems". Controlled release matrix dosage forms can include an insoluble matrix, a soluble matrix or a combination of insoluble and soluble matrices in which the rate of release is slower than that of an uncoated non-matrix conventional or immediate release dosage forms or uncoated normal release matrix dosage forms. Controlled release matrix dosage forms can be coated with a "control-releasing coat" to further slow the release of the bupropion salt from the controlled release matrix dosage form. Such coated controlled release matrix dosage forms can exhibit "modified-release", controlled-release", "sustained- release", "extended-release", "prolonged-release", "delayed- release" or combinations thereof of the bupropion salt.

The beta-alanine in the methods provided herein can be administered in any controlled release or sustained release dosage form or unit dosage form. Sustained release technologies include, but are not limited to, physical systems and chemical systems. Physical systems include, but are not limited to, reservoir systems with rate-controlling membranes such as encapsulation (e.g., micro- and macro-) and membrane systems; reservoir systems without rate-controlling membranes such as hollow fibers, ultra microporous cellulose triacetate, and porous polymeric substrates and foams; monolithic systems including those systems physically dissolved in non-porous, polymeric, or elastomeric matrices (e.g., non- erodible, erodible, environmental agent ingression, and degradable) and materials physically dispersed in non-porous, polymeric, or elastomeric matrices (e.g., non-erodible, erodible, environmental agent ingression, and degradable); laminated structures including reservoir layers chemically similar or dissimilar to outer control layers; and other physical methods such as osmotic pumps or adsorption onto ion-exchange resins.

Chemical controlled release systems include, but are not limited to, chemical erosion of polymer matrices (e.g., heterogeneous or homogeneous erosion) or biological erosion of a polymer matrix (e.g., heterogeneous, or homogeneous). Hydrogels also can be used as controlled release dosage forms (e.g., see Controlled Release Systems: Fabrication

Technology, Vol. II, Chapter 3; pages 41-60; "Gels For Drug Delivery").

There are a number of sustained release drug formulations that have been developed. These include, but are not limited to, microencapsulated powders; osmotic pressure- controlled gastrointestinal delivery systems; hydrodynamic pressure-controlled

gastrointestinal delivery systems; membrane permeation-controlled gastrointestinal delivery systems, which include microporous membrane permeation-controlled gastrointestinal delivery devices; gel diffusion controlled gastrointestinal delivery systems; and ion- exchange-controlled gastrointestinal delivery systems, which include cationic and anionic drugs. In some embodiments, the methods provided herein administered beta-alanine in a sustained release system that can include, e.g., an oil-microencapsulated sustained release powder dosage form that can be mixed with liquid and consumed as a drink mix beverage. See, also U.S. Pat. Nos. 5, 190,775; 6,013,286; 6,696,500; 6,756,049; 6,835,397; 6,919,372; 6,992,065; and 7,048,947.

In some methods, the beta-alanine is administered in a dosage form that includes an immediate release component and a controlled release component. For example, an immediate release layer containing beta-alanine can be coated onto the surface of substrates in which the beta-alanine is incorporated in a controlled release matrix. Where a plurality of the sustained release substrates containing an effective unit dose of the beta-alanine (e.g., multi-particulate systems including pellets, spheres or beads) are incorporated into a hard gelatin capsule, the immediate release portion of the beta-alanine dose can be incorporated into the gelatin capsule via inclusion of a sufficient amount of immediate release drug as a powder or granulate within the capsule. Alternatively, the gelatin capsule itself can be coated, on the interior or the exterior, with an immediate release layer of the beta-alanine.

In some embodiments, the beta-alanine can be incorporated into a food product. The food product may be a snack bar, a cereal, a dessert, including a frozen dessert, a functional food, or a drink. Non-limiting examples of a suitable drink include, e.g., fruit juice, a fruit drink, an artificially flavored drink, an artificially sweetened drink, a carbonated beverage, a sports drink, a liquid diary or dairy-like product or a shake, such as a ready-to-drink meal replacement beverage. In some embodiments, the beta-alanine is administered in a nutrition beverage or shake, such as a complete, balanced nutrition beverage or shake, or other liquid meal replacement beverage. The nutrition beverage or meal replacement beverage can be provided as a powder or granulated product which, when blended with an ingestible liquid, such as milk (e.g., cow's milk, soy milk or rice milk), juice or water, produces a highly palatable, highly nutritious instant shake or beverage. The beta-alanine also can be formulated in meal supplements, in enteral nutrition products and in parenteral nutrition products (e.g., such as described in Remington: The Science and Practice of Pharmacy (Gennaro, ed., Mack Publ, Co., Easton, PA (1995)). The beta-alanine also can be already mixed with an ingestible liquid and provided to a consumer already prepared and ready to consume. The beverage can include omega-3 amino acids.

The shake or nutritional beverage can include added vitamins and/or minerals. The advantages of vitamin and/or mineral fortification are now well understood. As the growing population continues to discover and become educated on the significant benefits of ensuring their diets contain essential vitamins and minerals such as calcium, magnesium, potassium and zinc, food and beverage manufacturers search for practical methods to ensure they provide adequate delivery methods. A positive correlation between calcium intake and bone mass has been found across many age groups. Magnesium is well understood in its role to maintain cardiovascular health, potassium for blood pressure maintenance and zinc for overall immune health.

The free beta-alanine dosage can be between about 1 mg and about 200 mg per kilogram body weight, or the dose of a biological source of beta-alanine (e.g., a peptide of beta-alanine, such as carnosine, or a salt of beta-alanine) can be between about 2.5 mg and about 500 mg per kg body weight. By way of example, suitable dosages for an 80 kg person per day can be between 0.08 grams to 16.0 grams of free beta-alanine or an amount of a biological source of beta-alanine that provides an equivalent amount of beta^alanine. In one aspect, the total amount of free beta-alanine administered in a controlled release dosage per day can be at least 200 mg, from 200 mg to 6.4 g, from 2.4 g to 12 g, or from 3.2 g to 16 g or more per day for a human. A single dose of active ingredient, e.g., free beta-alanine or a biological source thereof, can be formulated to be in the amount about 200 mg, 400 mg, 800 mg, 1, 200 mg, 1,400 mg, 1,600 mg, 2,400 mg, 3,200 mg, 4,800 mg, 6,400 mg or more.

Dosage amount, interval between doses, and duration of treatment can be adjusted to achieve a desired effect. In certain embodiments, dosage amount and interval between doses are adjusted to maintain a desired concentration of beta-alanine, or of increased carnosine synthesis, in a subject. For example, in certain embodiments, dosage amount and interval between doses are adjusted to provide plasma concentration of beta-alanine at an amount sufficient to achieve a desired effect, such as to increase the amount of carnosine in a muscle tissue. In certain of such embodiments the plasma concentration is maintained above the minimal effective concentration (MEC). Typically, a therapeutically effective dosage of beta- alanine should produce a blood, plasma or serum concentration of beta-alanine or an equivalent amount of a biological source of beta-alanine to muscle or other tissue of from about 0.1 μg/πύ to about 50-100 //g/ml. It has been determined that maintaining the serum levels of beta-alanine at an elevated level for a longer period of time is beneficial in the elderly.

In certain embodiments, the methods provided herein include administering beta-alanine with a dosage regimen designed to maintain a concentration above the MEC for 10-90% of the time, between 30-90% of the time, or between 50-90% of the time. In some methods, beta-alanine is administered at a dosage and over a period of time sufficient to increase muscle carnosine levels at least 10%, or at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110% , at least 120% , at least 130% , at least 140% , at least 150% or more. In some methods, beta- alanine is administered at a dosage and over a period of time sufficient to increase muscle carnosine levels to at least 30 mmol/kg dry muscle weight, or at least 32 mmol/kg dry muscle weight, or at least 34 mmol/kg dry muscle weight, or at least 36 mmol/kg dry muscle weight, or at least 38 mmol/kg dry muscle weight, or at least 40 mmol/kg dry muscle weight, or at least 42 mmol/kg dry muscle weight, or at least 44 mmol/kg dry muscle weight, or at least 46 mmol/kg dry muscle weight, or at least 48 mmol/kg dry muscle weight, or at least 50 mmol/kg dry muscle weight, or at least 52 mmol/kg dry muscle weight, or at least 54 mmol/kg dry muscle weight, or at least 56 mmol/kg dry muscle weight, or at least 58 mmol/kg dry muscle weight, or at least 60 mmol/kg dry muscle weight, or at least 65 mmol/kg dry muscle weight, or at least 70 mmol/kg dry muscle weight, or at least 75 mmol/kg dry muscle weight, or at least 80 mmol/kg dry muscle weight.

METHODS

Beta-alanine can be administered in methods to increase muscle mass, strength and physical function in a subject having or at risk of developing sarcopenia or frailty. In some embodiments, the method includes administering beta-alanine to a subject over a period of time, such as 1 week, 2 weeks, or more than 2 weeks, including 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 weeks, or for 30, 60, 90, 120, 180, 240 or more days, or for 1, 2, 3, 4, 5 or more years. The beta-alanine can be administered multiple times per day, such as 2, 3, 4, 5, 6, 7, or 8 times a day, or can be formulated for sustained or controlled release such that the beta-alanine can be administered in a dosage form once, twice or three or four times daily. The beta-alanine can be administered as a pharmaceutical with meals or between meals. The composition containing the beta-alanine can be administered any number of times that is both feasible and effective. The beta-alanine can be formulated in a food product such that it can be consumed as a part of a meal or as a snack.

The amount of beta-alanine per dose can be between 0.1 g and 16 g per day. In some embodiments, the beta-alanine is administered as a dosage of 200 mg, 400 mg, 600 mg, 800 mg, 1,000 mg, 1,200 mg, 1,400 mg, 1,600 mg, 1,800 mg, 2,000 mg, 2,400 mg, 2,800 mg, 3,200 mg, 4,000 mg, 4,400 mg, 4,800 mg, 5,400 mg, 5,800 mg or 6,400 mg one or more times a day. In some embodiments, the methods include administering a bolus dose or a sustained release or controlled release dosage or a combination thereof of beta-alanine to deliver a daily dosage of beta-alanine of between about 0.2 grams and 16 grams. In some embodiments, the daily dosage of beta-alanine is selected from among 200 mg, 250 mg, 400 mg, 500 mg, 600 mg, 650 mg, 750 mg, 800 mg, 1,000 mg, 1,200 mg, 1,250 mg, 1,400 mg, 1,500 mg, 1,600 mg, 1,650 mg, 1,750 mg, 1,800 mg, 2,000 mg, 2,200 mg, 2,250 mg, 2,400 mg, 2,500 mg, 2,600 mg, 2,650 mg, 2,750 mg, 2,800 mg, 3,000 mg, 3,200 mg, 3,250 mg, 3,400 mg, 3,500 mg, 3,600 mg, 3,650 mg, 3,750 mg, 3,800 mg, 4,000 mg, 4,200 mg, 4,250 mg, 4,400 mg, 4,500 mg, 4,600 mg, 4,650 mg, 4,750 mg, 4,800 mg, 5,000 mg, 5,200 mg, 5,250 mg, 5,400 mg, 5,500 mg, 5,600 mg, 5,650 mg, 5,750 mg, 5,800 mg, 6,000 mg, 6,200 mg, 6,250 mg, 6,400 mg, 6,500 mg, 6,600 mg, 6,650 mg, 6,750 mg, 6,800 mg, 7,000 mg, 7,200 mg, 7,250 mg, 7,400 mg, 7,500 mg, 7,600 mg, 7,650 mg, 7,750 mg, 7,800 mg, 8,000 mg, 8,200 mg, 8,250 mg, 8,400 mg, 8,500 mg, 8,600 mg, 8,650 mg, 8,750 mg, 8,800 mg, 9,000 mg, 9,200 mg, 9,250 mg, 9,400 mg, 9,500 mg, 9,600 mg, 9,650 mg, 9,700 mg, 9,750 mg, 9,800 mg, 9,850 mg, 9,900 mg, 9,950 mg and 10,000 mg.

In an alternative or further embodiment of the methods provided herein, the beta-^alanine can be administered in conjunction with exercise. For example, the beta-alanine can be administered prior to or following exercise. The beta-alanine can be administered, e.g., immediately prior to or immediately after exercise. In some methods, the exercise include aerobic exercise. In some embodiments, the exercise includes anaerobic exercise. In some embodiments, the method includes as a step performing a resistance exercise motion. In the methods provided herein, the beta-alanine can improve exercise tolerance in the elderly subject and/or minimize muscle fatigue.

Administration of a beta-alanine to a subject having or at risk of developing sarcopenia or frailty may stop or reverse a decline in skeletal muscle tissue mass, or muscle atrophy, when administered alone or in conjunction with exercise. In the methods provided herein, beta-alanine can increase muscle fitness in the elderly, alone or in combination with exercise, particularly in an elderly subject having or at risk of developing sarcopenia or frailty. Administration of beta-alanine also can increase physical endurance and physical performance in the elderly, particularly an elderly subject having or at risk of developing sarcopenia or frailty. In the methods provided herein, elderly subjects to which the beta- alanine is administered can perform physical activities for a longer time than elderly subjects not administered beta-alanine.

Administration of a beta-alanine to a subject having or at risk of developing sarcopenia or frailty also can improve skeletal muscle endurance and/or resistance to fatigue. Administering beta-alanine to an elderly subject can increase muscle oxidative capacity. Muscle oxidative capacity is a factor for muscle endurance and muscle fatigue resistance. Hence, this in combination with the increase in buffering capacity in muscle tissue afforded by increased carnosine synthesis upon administration of beta-alanine augments resistance to muscle fatigue in the elderly.

Administration of a beta-alanine to a subject having or at risk of developing sarcopenia or frailty can inhibit muscle catabolism and/or increase muscle anabolism, particularly when administering in combination with essential amino acids or with a protein source. In some methods, the beta-alanine is administered alone or in combination with a protein source, or in combination with one or more essential amino acids, or in combination with a protein and free essential amino acids.

Administration of a beta-alanine to a subject having or at risk of developing sarcopenia or frailty can improve the muscle:fat ratio in mammals including humans. In some embodiments, such methods include administering beta-alanine as a pharmaceutical dosage, dietary supplement or in the form of a foodstuff containing beta-alanine. The methods can include exercise as a step. The exercise can be aerobic, anaerobic or a combination thereof. The methods can include modifying the diet of the subject to increase the intake of protein by the subject. In some embodiments, the method includes co-administering beta-alanine with a dietary protein, such as an animal protein (for example milk, meat, fish or egg protein), a vegetable protein (for example soy, wheat, rice, bean or pea protein), a whey protein, an isolated whey protein, a soy protein, an isolated soy protein, a casein protein, or any combination thereof. In some embodiments, the methods include administering compositions containing one or more free essential amino acids. In some embodiments, the methods include modifying the caloric intake of the mammal by providing a diet high in protein and fiber but low in carbohydrates with a high glycemic index. In some methods, the fiber is soluble fiber, while in other methods, the fiber is insoluble fiber or a combination of soluble and insoluble fiber.

Administration of a beta-alanine to a subject having or at risk of developing sarcopenia or frailty can improve the gait of the subject, e.g., increase stride length, reduce stride frequency and/or reduce stance width variability, and also prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia. The increase in stride length following administration of the beta-alanine can be any increase, e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98%, 99% or more, relative to the stride length prior to treatment with the beta-alanine. In a particular embodiment, the stride length following administration of the beta-alanine is greater than the standardized average stride length.

Also provided are methods of decreasing stride frequency in a subject having sarcopenia or susceptible of developing sarcopenia, which include administering beta^alanine to the subject in an amount and for a sufficient time to decrease stride frequency. The decrease in stride frequency following administration of the beta-alanine can be any decrease, e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98%, 99% or more, relative to the stride frequency prior to treatment with the beta-alanine. Also provided are methods of decreasing stride length variability in a subject having sarcopenia or susceptible of developing sarcopenia, which include administering a beta- alanine to the subject in an amount and for a sufficient time to decrease stride length variability. The decrease in stride length variability following administration of the beta- alanine can be any decrease, e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98%, 99% or more, relative to the stride length variability prior to treatment with the beta-alanine.

Also provided are methods of increasing hand grip strength in a subject having or susceptible of developing sarcopenia or frailty, by administering a beta-alanine to the subject in an amount and for a sufficient time to increase hand grip strength. The increase in hand grip strength following administration of the beta-alanine can be any increase, e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98%, 99% or more, relative to the hand grip strength prior to treatment with the beta-alanine.

functionality of the subject. The methods can result improved results in the timed get-up-and- go test, the timed stand test, the stair climb muscle power test, and improvements in balance, such as the one-leg balance test or improvements in one or more criteria of the Berg balance test.

Also provided are methods of improving a Berg Balance test score in an elderly subject, the method including administering to the subject a beta-alanine in an effective amount and for sufficient time to improve the Berg Balance test score of the subject compared to a baseline score prior to administration of the beta-alanine. In the methods provided herein, the Berg Balance test score can be improved by at least +5, and can be improved by +6, +7, +8, +9, +10, +11, +12, +13, +14, +15 or more. In some methods, the score is improved to be in a range between 30 and 40 or between 40 and 50.

Also provided are methods of enhancing the physical capacity of a subject involved in muscular exertion by administering to the mammal a beta-alanine. It has been determined that administering a beta-alanine to subject having or susceptible of developing sarcopenia or frailty can enhance the subject's activity, e.g., exercise capacity. Exercise capacity can be limited by the rate by which oxygen can be taken up by a subject. When beta-alanine is administered to an elderly subject, the oxygen uptake, including the rate of oxygen uptake (aerobic metabolism) as well as anaerobic metabolism (oxidation of sugar in the absence of oxygen to produce lactic acid or lactate) can be enhanced relative to a state where beta- alanine is not administered. For example, it has been determined that when a beta-alanine is administered to a mammal as described herein, the VO2 level achieved is higher than when beta-alanine is not ingested. For example, the uptake of oxygen is higher when an individual participated in an exercise regimen, as described herein, and ingested beta-alanine in the amounts described herein and for a period of time as described herein than an individual that has not ingested beta-alanine.

A beta-alanine can be used to prevent, treat, delay, mitigate, decrease and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia in a subject. A beta-alanine can be used to prepare a medicament to prevent, treat, delay, mitigate, decrease and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia in a subject.

A beta-alanine also can be used for maintaining or increasing muscle mass and/or muscle strength and/or muscle function in a subject having or susceptible of developing sarcopenia or frailty. A beta-alanine also can be used to prepare a medicament for maintaining or increasing muscle mass and/or muscle strength and/or muscle function in a subject having or susceptible of developing sarcopenia or frailty.

A beta-alanine also can be used to improve an appendicular skeletal muscle mass t- score of a subject, particularly in a subject having a negative appendicular skeletal muscle mass t-score (a score less than zero). In some embodiments, the appendicular skeletal muscle mass t-score of a subject can be improved by +0.5, +1, +1.5, +2, +2.5, +3 or more.

A beta-alanine also can be used to stop or reverse a decline in skeletal muscle tissue function in a subject having or at risk of developing sarcopenia or frailty. A beta-alanine also can be used to prepare a medicament for stopping or reversing a decline in skeletal muscle tissue function in a subject having or at risk of developing sarcopenia or frailty.

A beta-alanine can be used to inhibit muscle catabolism and/or increase muscle anabolism in a subject having or at risk of developing sarcopenia or frailty. A beta-alanine can be used to prepare a medicament for inhibiting muscle catabolism and/or increasing muscle anabolism in a subject having or at risk of developing sarcopenia or frailty.

A beta-alanine can be used to improve the muscle:fat ratio in a subject having or at risk of developing sarcopenia or frailty. A beta-alanine can be used to prepare a medicament for improving the muscle:fat ratio in a subject having or at risk of developing sarcopenia or frailty.

A beta-alanine can be used to improve the gait of a subject having or at risk of developing sarcopenia or frailty. In some embodiments, improving the gait of the subject includes increasing stride length, reducing stride frequency, reducing stance width variability or a combination thereof. A beta-alanine can be used to prepare a medicament for improving the gait of a subject having or at risk of developing sarcopenia or frailty.

A beta-alanine can be used to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of frailty in a subject. Use of a beta-alanine can stop or reverse declines in functional reserve, reduce time to exhaustion, increase mean nominal walking speed, decrease muscle weakness and improve muscle functionality. Use of a beta-alanine can also lead to sustained or increased physical activity in a subject. A beta- alanine can be used to prepare a medicament to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of frailty in a subject.

A beta-alanine can be used to improve muscle functionality of a subject having or at risk of developing sarcopenia or frailty. The improvement in muscle functionality can be demonstrated by a reduction in the time required to complete a timed get-up-and-go test, or by a reduction in the time required to complete a timed stand test. A beta-alanine can be used to prepare a medicament to improve muscle functionality of a subject having or at risk of developing sarcopenia or frailty.

A beta-alanine can be used to improve a Berg Balance test score in an elderly subject. In some embodiments, a beta-alanine can improve a Berg Balance test score by at least +5. In some embodiments, a beta-alanine can improve a Berg Balance test score so that it is in a range between 30 and 40 or between 40 and 50. A beta-alanine can be used to prepare a medicament to improve a Berg Balance test score in an elderly subject.

A beta-alanine can be used to increase hand grip strength in a subject having or susceptible of developing sarcopenia or frailty. A beta-alanine can increase hand grip strength by any increment, e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98%, 99% or more, relative to the hand grip strength prior to treatment with the beta-alanine. A beta-alanine can be used to prepare a medicament to increase hand grip strength in a subject having or susceptible of developing sarcopenia or frailty.

A beta-alanine can be used to decrease stride frequency in a subject having sarcopenia or susceptible of developing sarcopenia. The decrease in stride frequency following administration of a beta-alanine can be any decrease, e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98%, 99% or more, relative to the stride frequency prior to treatment with the beta-alanine. A beta-alanine can be used to prepare a medicament to decrease stride frequency in a subject having sarcopenia or susceptible of developing sarcopenia.

A beta-alanine also can be used to decrease stride length variability in a subject having sarcopenia or susceptible of developing sarcopenia. The decrease in stride length variability following administration of the beta-alanine can be any decrease, e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98%, 99% or more, relative to the stride length variability prior to treatment with the beta-alanine.

When preparing any of the medicaments described herein, the amount of a beta- alanine can vary, and can be provided in a dosage effective to maintain a plasma

concentration of beta-alanine above the minimal effective concentration (MEC). The medicaments can be formulated as unit dosages. The amount of beta-alanine per unit dose can be between 100 mg and 2,000 mg, and a single unit dose can include a beta-alanine at a dose of 200 mg, 400 mg, 600 mg, 800 mg, 1,000 mg, 1,200 mg, 1,400 mg, 1,600 mg, 1,800 mg, 2,000 mg, 2,400 mg, 2,800 mg, 3,200 mg, 4,000 mg, 4,400 mg, 4,800 mg, 5,400 mg, 5,800 mg or 6,400 mg. A single bolus dose of beta-alanine greater than 800 mg can result in symptoms of paraesthesia. In some embodiments, single unit doses containing greater than 800 mg of a beta-alanine can be formulated as controlled release dosages. The medicament can be formulated so that it delivers a dosage of a beta-alanine over a period of 6 hours, 8 hours, 12 hours or 24 hours a total dose of a beta^alanine of between 0.1 g and 16 g per day. In some embodiments, the medicament can be formulated to deliver an amount of a beta- alanine of 200 mg, 400 mg, 600 mg, 800 mg, 1,000 mg, 1,200 mg, 1,400 mg, 1,600 mg, 1,800 mg, 2,000 mg, 2,400 mg, 2,800 mg, 3,200 mg, 4,000 mg, 4,400 mg, 4,800 mg, 5,400 mg, 5,800 mg or 6,400 mg.

I. Examples

The following examples are included for illustrative purposes only and are not intended to limit the scope of the embodiments provided herein.

EXAMPLE 1

12 week supplementation study

The effect of 12 weeks of dietary supplementation with beta-alanine in elderly subjects to assess effect on exercise tolerance and muscle carnosine concentration by MRS was evaluated. The treatment consisted of administering two 800 mg beta-alanine controlled release supplements 2 times per day (3.2 g beta-alanine in a 24 hour period), over the course of twelve weeks. Muscle carnosine levels were measured non-invasively during this time by MRS and the effect on exercise capacity, aerobic and/or anaerobic, was assessed using a treadmill test adapted for use with elderly people and with a pre-supplementation endurance time of approximately 4 minutes. In addition, "timed stands" and "timed up-and-go" tests were used to assess any changes in daily-life activity functionality. Additional testing included hand grip strength, gait measurements, including stride length, stride frequency and stance variability, V02peak and muscle to fat ratio to determine lean mass of the subject.

Elderly male subjects aged 60 to 80 years were recruited into the study.

Subjects were excluded:

1) where it was evident that joint disease or other causes of limited mobility would prevent the subject undertaking the exercise tests;

2) where subjects had been diagnosed with severe hypertension that had not been treated;

3) where subjects had metallic inserts as these would not be able to undertake the MRS measurements; and

4) where subjects felt anxious over the taking of the MRS scans.

Subjects were screened using a preliminary examination by a rheumatologist to approve their competency to participate in the study. Any history of diabetes, vascular, cardiac or respiratory problems, or other underlying medical problem was compiled for each subject.

GROUPS

The study was divided into two groups. In the first group, 12 elderly subjects were supplemented with beta-alanine. In the second group, 6 elderly subjects were supplemented with placebo. The group supplemented with beta-alanine were administered two 800 mg tablets containing Carnosyn® beta-alanine (Natural Alternatives International Inc., San Marcos, CA USA) two times per day for 12 weeks. The group supplemented with placebo were administered two 800 mg tablets containing placebo two times per day for 12 weeks.

MRS measurements of muscle carnosine were performed on the gastrocnemius muscle at 0, 6 and 12 weeks. An exercise capacity test was performed at the same times. Each subject completed an activity questionnaire at 0, 6 and 12 weeks. MRS MEASUREMENT OF MUSCLE CARNOSINE:

Muscle carnosine content was measured in vivo by ^-MRS using a whole body 3.0T MRI scanner (Achieve Intera, Philips, Best, The Netherlands) and a 14 cm diameter ^lH- surface coil. For the measurements, the surface coil was placed centered under the calf muscle of the left leg. The scanner body coil was used to obtain conventional anatomical Tl- weighted magnetic resonance images in the three orthogonal planes. Spectrum raw data then was analyzed with Java Magnetic Resonance User Interface software, and processing steps included apodization to 5Hz, Fourier transform and phase correction. The carnosine signal was quantified relative to an external reference.

PHYSICAL CAPACITY TESTS:

Subjects in each of the groups were assessed using physical capacity tests at 0, 6 and 12 weeks. These included exercise tests and muscle function testing.

Anaerobic Exercise Test

The subjects were required to visit the laboratory on 2 occasions within a period of one week. At the first visit, subjects performed an incremental test on a motorized treadmill to determine the ventilatory anaerobic threshold (VAT) and V0_2peak. The subjects also were submitted to one repetition square-wave transition from rest to exercise intensity

corresponding to 75% (Delta), i.e., 75% of the difference between VAT and V0_2peak, to the limit of tolerance (TLIM). The test is similar to the cycle ergometer test described by Puente- Maestu et al. (Respiration 70: 367-370 (2003)).

Muscle Function Tests

Assessment of muscle function was performed through "timed-stand" tests (i.e., the time needed to stand 10 times from a standard chair) and "timed-up-and go" tests (the time required for subjects to rise from a standard arm chair, walk to a line on the floor 3 meters away, turn, return, and sit down again), which has been previously validated to measure improvements in daily-life activities.

ACTIVITY QUESTIONNAIRE

Each subject completed an activity questionnaire at 0, 6 and 12 weeks to evaluate daily levels of physical activity before and during the study. The questionnaire is similar to that described in the art (e.g., see Voorips et al, Med Sci Sports Exerc 23: 974-979 (1991)). FOOD INTAKE

Food intake was assessed at 0, 6 and 12 weeks by three 24-hour dietary recalls undertaken on separate days (2 weekdays and 1 weekend day) using a visual aid photo album of real foods. The 24-hour dietary recall included the listing of foods and beverages consumed during the 24 hour period prior to the recall. Energy and macronutrient intakes were analyzed using nutritional computer software (e.g., Virtual Nutri (Philippi et al, Virtual Nutri (software)— Version 1.0 for Windows. Sao Paulo: Departamento de Nutricao da Faculdade de Saiide Piiblica da Universidade de Sao Paulo, (1996)).

DEMOGRAPHIC AND LABORATORY DATA

Measurements of height, weight and body mass index were assessed for each subject. The subjects also were given a complete physical examination performed by a

Rheumatologist. V0_2peak, VAT, time-to-exhaustion were determined through VO₂ testing. Serum and urinary creatinine, proteinuria, albuminuria, liver enzymes, and hematology were assessed.

BLOOD TESTS AND ECG

A blood sample for clinical biochemistry (liver, muscle, kidney function tests) and full hematology was taken at 0 and 12 weeks from all subjects. A 12 lead ECG was performed on each subject at 0 and 12 weeks.

STATISTICAL ANALYSIS

Data were tested by unpaired Student's t-test for delta changes. Data are expressed as mean ±

SD, except when otherwise stated.

RESULTS

1) All subjects were physically inactive (as assessed by the physical activity questionnaire) and well-nourished (as assessed by the subject global assessment) at baseline and throughout the protocol. Table 1 presents the subjects' demographic characteristics. Table 1. Subjects' demographic characteristics at baseline and after 12 weeks following either beta-alanine or placebo supplementation. Results include data from subject 6.

Beta-alanine (n=12) Placebo (n=6)

Pre Week 12 Pre Week 12

Gender (M7F) 6/6 2/4

Age (yrs) 65 ± 4 64 ± 7

Height (cm) 162 + 11 162 ± 5

Body mass (kg) 78.2 + 1 1.8 78.2 ± 11.2 74.7 ± 12.4 74.9 ± 1 1.1 BMI (Kg/cm²) 29.6 ± 2.7 29.6 ± 2.6 28.4 ± 4.3 28.4 ± 3.9 Body fat (kg) 25.9 ± 4.1 25.5 ± 3.7 26.1 ± 6.8 25.8 ± 6.3 Body fat (%) 33.4 + 5.2 34.0 + 5.0 34.8 + 8.2 34.3 + 7.2 LBM (kg) 50.1 + 9.7 50.6 + 9.8 47.1 + 10.1 47.5 + 9.5 BMD (g/cm²) 1.1 + 0.1 1.1 + 0.1 1.0 + 0.1 1.0 + 0.1

BMC (kg) 2.1 + 0.4 2.1 + 0.4 2.0 + 0.2 2.0 + 0.2

Abbreviations: M = males; F = females; BMI = body mass index; LBM = lean body mass; BMD = body mineral density; BMC = body mineral content. Data are expressed as mean + SD. There were no significant differences within or between groups.

Fig. 1 is a graph showing individual data for muscle carnosine content (arbitrary units) at baseline (PRE) and after 12 weeks of beta-alanine supplementation following either beta-alanine (Panel A, n = 10) or placebo supplementation (Panel B, n = 6). The highlighted line represents data from subject 6. Two MRS scans presented poor quality and their respective subjects were removed from analyses.

Fig. 2 is a graph showing individual data for absolute change in muscle carnosine content (arbitrary units) from baseline (PRE) to 12-week supplementation period (POST- 12). The value from subject 6 is shown circled. Two MRS scans presented poor quality and their respective subjects were removed from analyses. Both subjects were in the active treatment group. B-alanine n = 10; Placebo n = 6.

Fig. 3 is a graph showing time-to-exhaustion in the submaximal exercise test (TLIM; i.e., intensity corresponding to 75% of the difference between ventilatory threshold and V02 peak) at baseline (PRE) and after 12 weeks of beta-alanine supplementation (POST-12). * denotes repeated measures ANOVA within-group time effect, p = 0.04 for delta change in beta-alanine group vs. placebo group (following exclusion of subject 6, p = 0.03). Data are mean ± SE.

Fig. 4 is a graph showing correlation between percent change in the time-to- exhaustion in the TLIM test (i.e., intensity corresponding to 75% of the difference between ventilatory threshold and V02 peak) and the percent change in the muscle carnosine content. Light diamonds represent placebo-supplementation and dark diamonds represent beta- alanine-supplemented subjects.

Fig. 6 is a graph showing correlation between percent change in the time-to- exhaustion in the incremental test and the percent change in the muscle carnosine content. Light diamonds represent placebo-supplementation and dark diamonds represent beta- alanine-supplemented subjects.

2) MRS scans from two subjects were of such poor quality that they were deleted from analysis of the changes in muscle carnosine. Poor quality images can result for a number of reasons, including if the subject moves. Both subjects were subsequently found to have been in the beta-alanine supplementation group, leaving data of 10 to be analyzed.

One subject (subject 6) failed to show any apparent response to beta-alanine supplementation after the 12 weeks of supplementation. This subject, one of 10, nonetheless confirmed that they had taken the full course of treatment. Data collected at 6 weeks showed a small increase in muscle carnosine but this was later reversed at 12 weeks. It is possible that the decline at 12 weeks was due to an increase in muscle water or in the standing-blood volume in the voxel space, in which case adjustment of the carnosine values to the muscle total creatine signal obtained in the same scan would correct for these changes. For the present report, however, the data are presented both in full and also with this subject excluded as an outlier.

3) Supplementation with beta-alanine for 12 weeks significantly increased the gastrocnemius muscle carnosine content:

- in all elderly subjects by a mean of 72.4% (effect size: 1.06; delta change in beta- alanine group vs. placebo group: p = 0.038 (Student's t-test for unpaired data); Figures 1 and 2).

- following exclusion of subject 6 the mean increase was 85.4% (effect size: 1.21 ; delta change in beta-alanine group vs. placebo group: p = 0.004 (Student's t-test for unpaired data)).

4) Beta-alanine supplementation improved performance in the TLIM test by increasing the time-to-exhaustion at 12 weeks:

- in all elderly subjects by a mean of 33.2% (effect size: 1.52; delta change in beta- alanine group vs. placebo group: 0.07 (Student's t-test for unpaired data));

- following exclusion of subject 6 the mean increase was 36.5% (effect size: 1.71 ; delta change in beta-alanine group vs. placebo group: p = 0.05 (Student's t-test for unpaired data)). 5) There was a significant correlation (p = 0.03) between the percent change in time- to-exhaustion in the TLIM performance test and the percent change in muscle carnosine (Figure 4; Table 2). N.B. Subject 6 is included in Figure 4.

6) Beta-alanine supplementation improved performance in the incremental test by increasing the time-to-exhaustion at 12 weeks:

- in all elderly subjects by a mean of 10.6% (effect size: 0.89; delta change in beta- alanine group vs. placebo group: p = 0.06 (Student's t-test for unpaired data), whereas no change occurred in Placebo group (Figure 5)).

- following exclusion of subject 6 the mean increase was 12.2% (effect size: 1.03; delta change in beta-alanine group vs. placebo group: p = 0.04 (Student's t-test for unpaired data)).

7) There was a significant correlation (p = 0.024) between percent change in muscle carnosine and percent change in time-to-exhaustion in the incremental TTE (Figure 6; Table 2)·

Table 2. Correlations between delta changes in the performance tests vs. the changes in muscle carnosine content.

Relative Change in muscle carnosine (Δ%)

Relative change in TLIM (Δ%)

n = 16

P 0.003 r 0.656 r² 0.430

Relative change in TTE (Δ%)

n = 16

P 0.024

R 0.501

2

r 0.251

Abbreviations: TLIM = time-to-exhaustion in a supramaximal exercise test (i.e., intensity corresponding to 75% of the difference between ventilatory threshold and VO₂ peak); TTE = time-to-exhaustion in an incremental test.

8) There were no significant changes for body mass (Table 1), food intake (Table 3), quality of life (Table 4), muscle strength and muscle function (Table 5), and cardiopulmonary parameters (Table 6). Table 3. Food intake at baseline, after 6 weeks and after 12 weeks following either beta- alanine or placebo supplementation.

Beta-alanine (n=12) Placebo (n= =6)

Pre Week 6 Week 12 Pre Week 6 Week 12

Energy 2263 ± 2404 ± 2068 ± 2184 + 2247 ± 2083 ±

(Kcal/d) 332 251 513 409 336 352

Carbohydrate

% of energy 45 ±7 47 ± 1 44 ±5 54 ±5 55 ±0 46 ± 1 g/d 257 ±55 281 ±33 228 ±51 298 ±23 305 ±41 238 ± 35

Fat

% of energy 38 ± 5 34 ± 1 36±4 31 ±5 31±2 36± 1 g/d 96 ± 17 91 ±10 83 ±20 76 ±26 76 ±6 84 ± 15

Protein

% of energy 16 ± 3 19 ± 1 20 ±3 14+1 15 ±2 18± 1 g/d 93 ±25 116 ± 11 103 ±44 77 ±20 85 ±29 94 ± 19

Data are expressed as mean ± SD. There were no significant differences within or between groups.

Table 4. Short Form Health Survey (SF36) data at baseline, after 6 weeks and 12 weeks following either beta-alanine or placebo supplementation.

Beta-alanine (n=12) Placebo (n=6)

Domain Pre Week 6 Week 12 Pre Week 6 Week 12

Physical

74 ±9 61 ± 16 78 ±7 74 ±7 functioning

Physical role 16 ± 8 18 + 6 18 + 6 11 ±9 14 + 6 16 ± 5 functioning

Bodily pain 63 + 18 74 + 25 71 +20 59 + 13 60 ±20 61 ±20

General

health 77 + 14 77 + 11 82 + 11 70 + 13 71 ± 11 72 + 12 perceptions

Vitality 73 + 11 78 ± 12 78 ± 12 59 + 18 62+11 63 ± 17

Social role 89 + 16 94+11 91 ± 15 69 + 19 75 ±21 73 ±23 functioning

Emotional 21+7 21+7 22 + 2 15 + 7 12+11 14+11 role

functioning

Mental 79 + 15 83 ± 10 83 + 9 75 + 14 75 ± 10 81 ± 16 health

Data are expressed as mean + SD. There were no significant differences within or between groups.

Table 5. Strength and functional tests at baseline, after 6 weeks and 12 weeks following either beta-alanine or placebo supplementation.

Beta-alanine (n= =12) Placebo (n=6)

Pre Week 6 Week 12 Pre Week 6 Week 12

Leg Press (kg) 166.9 ± 170.3 ± 175.8 ± 148.0 ± 155.5 ± 155.2 ±

56.5 55.6 56.2 74.2 88.3 83.7

Bench press (kg) 32.3 ± 9.8 32.2 ± 32.5 ± 34.4 ± 34.8 ± 35.2 ±

10.0 10.1 16.3 16.9 16.5

Hand grip (kg) 34.0 ± 9.6 32.8 ± 32.8 ± 32.3 ± 8.4 32.4 ± 9 33.1 ±

10.1 10.6 8.3

Timed-stands 16.4 ± 2.2 17.1 ± 17.3 ± 15.3 ± 3.0 15.8 ± 16.3 ±

(reps) 1.9 1.6 2.8 2.7

Timed-up-and-go 6.3 ± 0.8 6.3 ± 0.7 6.2 ± 1.1 6.3 ± 1.2 6.2 ± 1.1 6.2 + 1.1

(s)

Endurance 12.9 ± 4.4 11.8 ± 14.1 ± 16.5 ± 4.6 13.8 ± 15.8 ± strength - set 1 4.2 3.5 3.7 3.1

(reps)

Endurance 8.3 ± 2.7 8.2 ± 2.6 9.8 ± 2.7 1 1.2 ± 4.2 1 1.8 + 12.5 ± strength - set 2 1.6 2.9

(reps)

Endurance 7.3 ± 1.9 6.3 ± 2.1 8.2 ± 2.6 9.5 ± 4.4 10.2 ± 1 1.5 ± strength - set 3 2.4 2.8

(reps)

Table 6. Cardiopulmonary data in the maximal incremental test at baseline and after 12 weeks following either beta-alanine or placebo supplementation. Beta-alanine (n=12) Placebo (n=6)

Pre Week 12 Pre Week 12

V0₂ rest 0.3+0.1 0.3 ±0.0 0.3 ±0.1 0.3 ±0.0 (L/min)

V0₂ peak 1.8 ±0.4 1.8 ±0.5 1.7 ±0.3 1.6 ±0.4

(L/min)

V0₂ peak 22.5 + 3.2 22.8 + 21.9 ±5.0 20.6 ±

(mL/Kg/min) 4.0 3.3

V0₂ AT 1.1+0.2 1.1+0.2 1.0 ±0.2 1.0 ±0.2 (L/min)

V0₂VT 1.5+0.3 1.5 + 0.3 1.5+0.3 1.5 + 0.4 (L/min)

RER peak 1.1 ±0.1 1.1 ±0.1 1.2 ±0.1 1.1 ±0.1

HR rest 74.2 + 76.4 ± 69.8 + 72.8 +

(bpm) 13.5 11.0 12.0 7.4

Abbreviations: VO₂ = oxygen consumption; AT = anaerobic threshold; VT = ventilatory threshold; RER = respiratory rate exchange; HR = heart rate. Data are expressed as mean

SD. There were no significant differences within or between groups.

9) Laboratory parameters indicated no adverse effects (Table 7). There were no reported side effects.

Table 7. Laboratory data at baseline, after 6 weeks and 12 weeks following either beta- alanine or placebo supplementation.

Beta-alanine (n=12) Placebo (n=6)

Pre Week 6 Week 12 Pre Week 6 Week 12

Aldolase (U/L)

3.4 + 0.9 3.9 ± 1.6 3.0+1.2 4.0 ±0.3 4.2 ± :0.7 4.1 ±0.7

ALT (U/L) 28.2 + 26.7 ± 23.2 ± 23.0 ±

28.7 ± 15.6 19.0: + 5.1

15.0 7.5 6.6 10.9

AST (U/L) 24.3 + 22.6 ± 23.3 ±

24.9 ±8.5 21.1 : ±2.3 21.8±4.0

7.0 4.7 2.1

GGT (U/L) 29.00 + 32.08 ± 28.09 ± 17.83 ± 15.60 4.93 15.50 ± 8.71 14.11 7.54 5.91 5.82

Total bilirubin

(mg/dL) 0.7 + 0.2 0.7 + 0.3 0.6 + 0.2 0.6 + 0.1 0.5 + 0.1 0.6 + 0.2

CK (U/L) 141.7 + 149.5 ± 123.9 + 117.0 ± 118.2 + 148.0 ±

82.6 117.7 65.2 87.7 57.7 107.0

Creatinine

(mg/dL) 0.9 + 0.2 0.9 + 0.2 0.9 + 0.1 0.9 + 0.1 0.8 + 0.2 0.83 + 0.2

Glycemia (mg/dL) 109.7 114.4 + 89.0 +

105.7 + 16.5 93.3 + 5.9 95.0 + 8.4

15.7 17.9 9.0

LDH (U/L) 334.4 : 323.2 + 385.0 : 407.0 ; 401.7 ±

356.8 + 28.8

34.4 28.9 35.7 12.5 52.3

Urea (mg/dl) 37.58 ; 38.45 + 35.67 ; 39.83 ;

36.17 + 7.38 36.67 9.46 8.13 7.09 6.77 4.75

Erythrocytes (1

4.94 + 4.94 + 4.82 + 4.82 + 4.80 ± million/mm3) 4.99 + 0.28

0.34 0.31 0.45 0.48 0.53

Hemoglobin

15.05 : 14.85 15.05 ; 14.90 ; 14.72 + (g/dL) 15.21 + 0.92

1.31 1.09 0.92 1.03 1.09

Hematocrit (%) 44.33 ; 44.63 : 44.30 ; 45.42 ; 44.37 ±

46.18 + 2.93

3.20 2.50 3.12 3.29 3.17

Leukocytes

7.4 + 1.6 7.4 + 2.4 7.5 + 2.7 6.1 + 1.0 6.8 + 2.0 6.6 + 1.5

(1000/mm³)

Neutrophils

3.9 + 1.2 3.8 + 1.4 3.7 + 1.5 3.0 + 0.6 3.5 + 1.6 3.7 + 1.5

(1000/mm³)

Eosinophils

0.2 + 0.1 0.4 + 0.6 0.5 + 1.0 0.3 + 0.2 0.3 + 0.3 0.2 + 0.2

(1000/mm³)

Basophils 0.01 + 0.02 + 0.02 + 0.03 + 0.02 +

0.03 + 0.05

(1000/mm³) 0.03 0.04 0.04 0.05 0.04

Lymphocytes

2.6 + 0.5 2.5 + 0.6 2.6 + 0.8 2.2 + 0.6 2.3 + 0.7 2.1 + 0.8

(1000/mm³) Monocytes

0.6 ± 0.2 0.6 + 0.2 0.6 + 0.2 0.6 + 0.1 0.7 + 0.1 0.6 + 0.1

(1000/mm³)

Platelets 266.5 ± 261.0 + 252.7 ; 253.0 ;

257.4 + 60.9

(1000/mm³) 59.8 61.3 35.8 46.3 67.2

Urinary

1.3 ± 0.6 1.2 + 0.6 1.2 ± 0.4 1.0 + 0.8 0.8 + 0.5 0.9 + 0.3 creatinine (g/L)

Microalbuminuria

6.3 ± 3.2 7.0 + 4.2 6.7 ± 3.4 6.8 + 6.5 6.8 + 4.2 4.5 + 3.6

(mg/L)

Proteinuria (g/L) 0.07 ± 0.08 ±

0.05 + 0.05

0.05 0.04 0.05 0.05 0.05

Data are expressed as mean ± SD. Abbreviations: AST = aspartate amino transferase; ALT alanine aminotransferase; LDH = lactate dehydrogenase; creatine kinase = CK; GGT = glutamyltransferase. There were no significant differences within or between groups.

Appendix 1. Intra-class reliability data of gastrocnemius carnosine content in two subjects submitted to serial spectrum acquisitions throughout one single day.

Car Water CLF Car/water Car*water Car*CLF*Water Car*CLF amplitude amplitude

Subject

A

Set 1 2.511 3.640 0.939 0.747 9.908 9.309 2.577

Set 2 2.511 3.510 0.940 0.715 8.813 8.285 2.360

Mean - - - 0.731 9.360 8.797 2.459

SD - - - 3.133 8.267 8.230 5.664

CV - - - 3.133 8.267 8.230 5.664

Subject

B

Set 1 4.191 3.480 0.963 1.204 14.584 14.058 4.039

Set 2 3.981 3.440 0.965 1.157 13.694 13.224 3.844

Set 3 4.172 3.440 0.950 1.212 14.351 13.644 3.966

Mean - - - 1.191 14.210 13.642 3.950

SD - - - 0.029 0.461 0.416 0.098

CV - - - 2.510 3.247 3.053 2.496 Abbreviations: Car = carnosine; CLF = coil loading factor. Overall, carnosine correction by water provides the more reliable data.

CONCLUSIONS

The report shows that 12-weeks of beta-alanine supplementation increases the muscle carnosine content in elderly people. The correlations performed in this study indicated positive associations between beta-alanine-induced muscle carnosine increase and improvements in exercise tolerance, showing that dietary supplementation with beta-alanine has a role in improving daily-physical activity and exercise capacity in elderly people.

EXAMPLE 2

Beta-alanine was administered to an active 66 year old male subject over a period of

12 weeks. The subject is an avid participant in a number of fitness activities, including fast up-hill walking, swimming and cycling, and has participated in these activities for a number of years. The subject's self-assessed performance in these activities was that his performance in these activities was constant but declining slightly in recent years.

The subject was administered 800 mg beta-alanine formulated in a controlled release tablet 3 times daily for a period of 12 weeks. The subject indicated that beta-alanine was taken every day of the study.

After 4 weeks of supplementation, the subject reported that he noticed improvements in his up-hill walking and sprint cycling activity. After about 8 weeks of supplementation, the subject reported that he experienced significant improvement in his day-to-day activities, including his sustained anaerobic athletic activities. As an example, he reported that his sustained front-crawl swimming speed improved. The subject reported that he noticed the greatest improvement in his fast up-hill walking ability. This exercise is anaerobically stressful to the gastrocnemius muscle). After 12 weeks of supplementation, without any change in training procedures or any increase in training frequency or schedules, the subject reported increases in muscle functionality, particularly in the execution of day-to-day activities. The subject reported that prior to beta-alanine supplementation, his athletic activities were consistent to slightly declining, and because the subject did not change the duration or intensity of his activities, the subject attributed the increases in athletic performance he experienced to the beta-alanine supplementation.

While various embodiments of the subject matter provided herein have been described, it should be understood that they have been presented by way of example only, and not limitation. Since modifications will be apparent to those of skill in this art, it is intended that this invention be limited only by the scope of the appended claims.

Claims

What is claimed:

1. A method to prevent, treat, delay, mitigate, decrease and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia in a subject, comprising:

administering to the subject a therapeutically effective amount of a beta-alanine, and thereby preventing, treating, delaying, mitigating and/or ameliorating the onset, advancement, severity and/or symptoms of sarcopenia in the subject.

2. A method of maintaining or increasing muscle mass and/or muscle strength and/or muscle function in a subject having or susceptible of developing sarcopenia or frailty, comprising:

administering to the subject a therapeutically effective amount of a beta-alanine and thereby increasing muscle mass and/or muscle function in the subject.

3. The method of claim 2, wherein the subject has an appendicular skeletal muscle mass t-score selected from among (a) <-3, (b) <-2.5, (c) <-2, (d) <-1.5, (e) <-1.0, and (f) <-0.5.

4. The method of claim 2, wherein the subject's appendicular skeletal muscle mass t-score is increased after at least 45 days of treatment, or is increased at least 90 days of treatment, or is increased after at least 180 days of treatment or is increased after at least one year of treatment.

5. The method of claim 4, wherein the appendicular skeletal muscle mass t-score is increased by at least 0.5 after treatment.

6. The method of claim 4, wherein the appendicular skeletal muscle mass t-score is increased by at least 1.0 after treatment.

7. A method of stopping or reversing a decline in skeletal muscle tissue function in a subject having or at risk of developing sarcopenia or frailty, comprising:

administering to the subject a therapeutically effective amount of a beta-alanine and thereby stopping or reversing a decline in skeletal muscle tissue function in the subject.

8. A method of inhibiting muscle catabolism and/or increasing muscle anabolism in a subject having or at risk of developing sarcopenia or frailty, comprising:

administering to the subject a therapeutically effective amount of a beta-alanine and thereby inhibiting muscle catabolism and/or increasing muscle anabolism in the subject.

9. A method of improving the muscle:fat ratio in a subject having or at risk of developing sarcopenia or frailty, comprising: administering to the subject a therapeutically effective amount of a beta-alanine and thereby improving the muscle:fat ratio in the subject.

10. A method of improving the gait of a subject having or at risk of developing sarcopenia or frailty, comprising:

administering to the subject a therapeutically effective amount of a beta-alanine and thereby improving the gait of the subject.

1 1. The method of claim 10, wherein improving the gait of the subject comprises increasing stride length, reducing stride frequency, reducing stance width variability or a combination thereof.

12. A method to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of frailty in a subject, comprising:

administering to the subject a therapeutically effective amount of a beta-alanine and thereby preventing, treating, delaying, mitigating and/or ameliorating the onset, advancement, severity and/or symptoms of frailty in the subject.

13. A method of improving muscle functionality of a subject having or at risk of developing sarcopenia or frailty, comprising:

administering to the subject a beta-alanine in an effective amount and for sufficient time to improve the muscle functionality of the subject.

14. The method of claim 13, wherein the improvement in muscle functionality is demonstrated by a reduction in the time required to complete a timed get-up-and-go test.

15. The method of claim 13 or 14, wherein the improvement in muscle functionality is demonstrated by a reduction in the time required to complete a timed stand test.

16. A method of improving a Berg Balance test score in an elderly subject, comprising:

administering to the subject a beta-alanine in an effective amount and for sufficient time to improve the Berg Balance test score of the subject compared to a baseline score prior to administration of the beta-alanine.

17. The method of claim 16, wherein the score is improved by at least +5.

18. The method of claim 16 or 17, wherein the score is improved to be in a range between 30 and 40 or between 40 and 50.

19. The method of any one of claims 1-18, wherein the age of the subject is selected from among (a) at least 40, (b) at least 50, (c) at least 55, (d) at least 60, (e) at least 65, and (f) at least 70 years of age.

20. The method of any one of claims 1-18, wherein the age range of the subject is selected from among (a) 40-50, (b) 50-60, (c) 60-70 and (d) greater than 70 years of age.

21. The method of any one of claims 1-20, wherein the subject has suffered some loss of muscle mass, but does not suffer from a condition that interferes with acts of daily living and/or prevents the subject from living an independent life.

22. The method of any one of claims 1-21, wherein the subject exhibits one or more symptoms of sarcopenia.

23. The method of any one of claims 1-22, wherein the subject exhibits one or more symptoms of frailty.

24. The method of any one of claims 1-23, wherein the beta-alanine is administered to the subject over a period of time of between 1 week and 1 year.

25. The method of any one of claims 1-23, wherein the beta-alanine is administered to the subject for at least 6, 8, 10, 12, 14, 16, 18 or 20 weeks.

26. The method of any one of claims 1-23, wherein the beta-alanine is administered to the subject for 1 year or more.

27. The method of any one of claims 1-26, wherein the beta-alanine is administered in a dose of between 0.1 g and 16 g per day.

28. The method of any one of claims 1-26, wherein the beta-alanine is administered as a dosage of 200 mg, 400 mg, 600 mg, 800 mg, 1,000 mg, 1,200 mg, 1,400 mg, 1,800 mg, 2,400 mg, 2,800 mg, 3,200 mg, 4,000 mg, 4,800 mg, or 5,400 mg one or more times a day.

29. The method of any one of claims 1-26, wherein the beta-alanine is administered as a bolus dosage or as a sustained release or controlled release dosage or a combination thereof, wherein the amount of beta-alanine delivered in a 24 hour period is between about 0.2 grams and 20 grams.

30. The method of any one of claims 1-29, wherein the beta-alanine is administered as a sustained release or controlled release dosage that delivers a dose of 2,400 mg, 2,800 mg, 3,000 mg, 3,200 mg, 3,600 mg, 4,000 mg, 4,200 mg, 4,400 mg, 4,800 mg, 5,000 mg, 5,200 mg or 5,400 mg beta-alanine.

31. The method of any one of claims 1-30, further comprising subjecting the subject to physical exercise.

32. The method of claim 31, wherein the exercise is aerobic exercise.

33. The method of claim 31, wherein the exercise is anaerobic exercise.

34. The method of any one of claims 31-33, wherein the beta-alanine is administered prior to the exercise.

35. The method of any one of claims 31-33, wherein the beta-alanine is administered after the exercise.

36. The method of any one of claims 31-35, wherein the exercise is resistance exercise.

37. The method of any of claims 1-36, wherein the beta-alanine is administered in a dosage effective to maintain a plasma concentration of beta-alanine above the minimal effective concentration (MEC).

38. The method of claim 37, wherein the dosage of beta-alanine is effective to maintain a plasma concentration of beta-alanine above the MEC for 10-90% of the time, or between 30-90% of the time, or between 50-90% of the time or between 20-80% of the time.

39. The method of any one of claims 1-38, wherein the beta-alanine is

administered at a dosage and over a period of time sufficient to increase muscle carnosine levels at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100%, or at least 110%, or at least 120%, or at least 130%, or at least 140%, or at least 150% or more from the level before administration of initial beta-alanine commenced.

40. The method of any one of claims 1-39, wherein the beta-alanine is

administered at a dosage and over a period of time sufficient to increase muscle carnosine levels to at least 30 mmol/kg dry muscle weight, or at least 32 mmol/kg dry muscle weight, or at least 34 mmol/kg dry muscle weight, or at least 36 mmol/kg dry muscle weight, or at least 38 mmol/kg dry muscle weight, or at least 40 mmol/kg dry muscle weight, or at least 42 mmol/kg dry muscle weight, or at least 44 mmol/kg dry muscle weight, or at least 46 mmol/kg dry muscle weight, or at least 48 mmol/kg dry muscle weight, or at least 50 mmol/kg dry muscle weight, or at least 52 mmol/kg dry muscle weight, or at least 54 mmol/kg dry muscle weight, or at least 56 mmol/kg dry muscle weight, or at least 58 mmol/kg dry muscle weight, or at least 60 minol/kg dry muscle weight.

41. The method of any of claims 1-40, wherein the beta-alanine is formulated for oral, rectal, transmucosal, intestinal, enteral, topical, transdermal, intrathecal, intraventricular, intraperitoneal, intranasal, or parenteral (e.g., intravenous, intramuscular, intramedullary, and subcutaneous) administration.

42. The method of any of claims 1-41, wherein the beta-alanine is administered in a controlled release formulation.

43. The method of claim 42, wherein the controlled release formulation comprises one or more of the following: a matrix composition, an encapsulating coating, an erosional system and a diffusional system.

44. The method of any of claims 1-43, wherein the beta-alanine is administered in a ready-to-drink meal replacement beverage.

45. The method of any of claims 1-43, wherein the beta-alanine is administered in an enteral nutrition product or in a parenteral nutrition product.

46. The method of any of claims 1-45, wherein the beta-alanine is co-administered with another agent.

47. The method of claim 46, wherein the agent co-administered with the beta- alanine is selected from among a creatine, a histidine, a nutraceutical, a vitamin, a mineral, an omega 3 fatty acid or combinations thereof.

48. The method of claim 47, wherein the creatine is creatine monohydrate or an ester, chelate, salt, amide, ether, hydrate, solvate or combination thereof.

49. The method of claim 46, wherein the agent co-administered with the beta- alanine is selected from among modulators of one or more of skeletal myosin, skeletal actin, skeletal tropomyosin, skeletal troponin C, skeletal troponin I, skeletal troponin T, and skeletal muscle, including fragments and isoforms thereof, and the skeletal sarcomere, anti-obesity agents, a vitamin D drug, orlistat, sibramine, diethylpropion, phentermine, benzaphetamine, phendimetrazine, estrogen, estradiol, levonorgestrel, norethindrone acetate, estradiol valerate, ethinyl estradiol, norgestimate, conjugated estrogens, esterified estrogens,

medroxyprogesterone acetate, testosterone, insulin-derived growth factor, human growth hormone, riluzole, cannabidiol, prednisone, albuterol, non-steroidal anti-inflammatory drugs, thyrotropin-releasing hormone (TRH), diethylstilbesterol, theophylline, enkephalins, E series prostaglandins, sulbenox, a growth hormone secretagogue, growth hormone releasing factor and its analogs, growth hormone, a growth hormone analog, a somatomedin, an alpha- adrenergic agonist, a serotonin 5-HTD agonist, sumatriptan, a somatostatin inhibitor, a somatostatin release inhibitor, parathyroid hormone, a bisphosphonate, estrogen, testosterone, a selective estrogen receptor modulator, a progesterone receptor agonists, an aP2 inhibitor, a PPAR gamma antagonist, a PPAR delta agonist, a beta 3 adrenergic agonist, a lipase inhibitor, a serotonin (and dopamine) reuptake inhibitor, an anorectic agents, indinavir sulfate, saquinavir, saquinavir mesylate, ritonavir, lamivudine, zidovudine, zalcitabine, lamivudine/zidovudine combinations, didanosine, stavudine, megestrol acetate, an anti- resorptive agent, vitamin D, vitamin D analogues, and cathepsin K inhibitors and

combinations thereof.

50. The method of claim 46, wherein the agent co-administered with the beta- alanine is selected from among a histidine, vitamin D, Vitamin C, Vitamin B l, Vitamin B2,

Vitamin B3, Vitamin B5, Vitamin B6, Vitamin B12, and/or Vitamin K, a mineral, such as chromium, iron, magnesium, sodium, potassium, vanadium, an amino acid, such as L- arginine, L-ornithine, L-glutamine, L-tyrosine, L-taurine, L-leucine, L-isoleucine, L-theanine and/or L- valine and derivatives thereof, one or more peptides, such as L-carnitine, carnosine, anserine, balenine, homocarnosine, kyotorphin, and/or glutathione and derivatives thereof, a methylxanthine, such as caffeine, aminophylline or theophylline, antioxidants, such as tocopherol, lutein, zeaxanthine, a flavanol, such as a flavanol extracted from tea or chocolate, and adenosine triphosphates.

51. A beta-alanine for use in preventing, treating, delaying, mitigating, decreasing and/or ameliorating the onset, advancement, severity and/or symptoms of sarcopenia in a subject.

52. Use of a beta-alanine to prepare a medicament to prevent, treat, delay, mitigate, decrease and/or ameliorate the onset, advancement, severity and/or symptoms of sarcopenia in a subject.

53. A beta-alanine for use in maintaining or increasing muscle mass and/or muscle strength and/or muscle function in a subject having or susceptible of developing sarcopenia or frailty.

54. Use of a beta-alanine to prepare a medicament for maintaining or increasing muscle mass and/or muscle strength and/or muscle function in a subject having or susceptible of developing sarcopenia or frailty.

55. A beta-alanine for use in improving an appendicular skeletal muscle mass t- score of a subject.

56. Use of a beta-alanine for preparing a medicament to improve an appendicular skeletal muscle mass t-score of a subject.

57. A beta-alanine for use in stopping or reversing a decline in skeletal muscle tissue function in a subject having or at risk of developing sarcopenia or frailty.

58. Use of a beta-alanine to prepare a medicament for stopping or reversing a decline in skeletal muscle tissue function in a subject having or at risk of developing sarcopenia or frailty.

59. A beta-alanine for use in inhibiting muscle catabolism and/or increasing muscle anabolism in a subject having or at risk of developing sarcopenia or frailty.

60. Use of a beta-alanine to prepare a medicament for inhibiting muscle catabolism and/or increasing muscle anabolism in a subject having or at risk of developing sarcopenia or frailty.

61. A beta-alanine for use in improving the muscle:fat ratio in a subject having or at risk of developing sarcopenia or frailty.

62. Use of a beta-alanine to prepare a medicament for improving the muscle:fat ratio in a subject having or at risk of developing sarcopenia or frailty.

63. A beta-alanine for use in improving the gait of a subject having or at risk of developing sarcopenia or frailty.

64. Use of a beta-alanine to prepare a medicament for improving the gait of a subject having or at risk of developing sarcopenia or frailty.

65. A beta-alanine for use in decreasing stride frequency in a subject having sarcopenia or susceptible of developing sarcopenia.

66. Use of a beta-alanine to prepare a medicament to decrease stride frequency in a subject having sarcopenia or susceptible of developing sarcopenia.

67. A beta-alanine for use in decreasing stride length variability in a subject having sarcopenia or susceptible of developing sarcopenia.

68. A beta-alanine for use in preventing, treating, delaying, mitigating and/or ameliorating the onset, advancement, severity and/or symptoms of frailty in a subject.

69. Use of a beta-alanine to prepare a medicament to prevent, treat, delay, mitigate and/or ameliorate the onset, advancement, severity and/or symptoms of frailty in a subject.

70. A beta-alanine for use in improving muscle functionality of a subject having or at risk of developing sarcopenia or frailty.

71. Use of a beta-alanine to prepare a medicament to improve muscle functionality of a subject having or at risk of developing sarcopenia or frailty.

72. A beta-alanine for use in improving a Berg Balance test score in an elderly subject.

73. Use of a beta-alanine to prepare a medicament to improve a Berg Balance test score in an elderly subject.

74. A beta-alanine for use in increasing hand grip strength in a subject having or susceptible of developing sarcopenia or frailty.

75. Use of a beta-alanine to prepare a medicament to increase hand grip strength in a subject having or susceptible of developing sarcopenia or frailty.