JP2010150263A - Method and composition for enhancing anaerobic working capacity in tissue - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for enhancing buffer performance of muscles and reducing fatigue of muscles by regulating accumulation of hydronium ions generated during continuous intense movement or movement continued under a locally low oxygen state. <P>SOLUTION: A composition comprising a β-alanylhistidine peptide and β-alanine, and glycine, insulin, an insulin-mimetic compound or an insulin action modifier, and a method for administering those peptides and amino acids are provided. The composition and method bring plasma concentration increase of β-alanine and/or creatinine and enhance anaerobic working capacity in tissues. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to the technical field of pharmaceuticals and physiological functions. In one embodiment, the present invention provides a method for increasing muscle buffer capacity and reducing muscle fatigue. The present invention also provides methods and compositions for enhancing the anaerobic working capacity of muscles and other tissues.

  Common dietary supplements are typically designed to compensate for reduced levels of nutrients in modern human and animal diets. In particular, useful dietary supplements enhance tissue function when ingested. Certain types of animals (eg, vegetarians and other animals that eat herbivorous foods) that may lack nutrients that can only be obtained from meat and animal foods in their normal diet Supplementing your diet can be particularly important.

  For example, in sports and athletic societies, regular dietary supplements that specifically improve athletic performance are increasingly important, such as dietary supplements that develop or enhance physical abilities for leisure or work purposes. It has become. In another example, anaerobic (eg, lactic acid production) stress may be sustained or intense exercise (eg, in soccer or ice hockey), such as oxygen availability may be limited (eg, peripheral vascular injury, free diving or synchronized swimming). Intermittent sprinting) and fatigue and distress that can be experienced by aging. Anaerobic stress can also result from long-term maximum isometric exercise when local blood flow is partially or totally occluded by increased intramuscular pressure (eg, during rock climbing). Excess lactic acid production can cause acidification of the intracellular environment.

  Creatine (ie, N- (aminoiminomethyl) -N-glycine, N-amidinosarcosine, N-methyl-N-guanylglycine, or methylglycosylamine) is characterized by its ability to meet high and variable energy demands. , Found in large quantities in skeletal muscle and other “excitable” tissues (eg, smooth muscle, heart muscle, or sperm). Creatine is converted to phosphoryl creatine in a biochemical pathway of energy generation within the cell. In mammalian skeletal muscle, the standard content including all of creatine (ie, creatine and phosphoryl creatine) ranges from less than 25 to about 50 mmol / kg fresh muscle (ie, 3.2-6.5 grams / kg fresh muscle). ).

  Creatine is formed in the liver and taken up into tissues such as muscle by an active transport system. Creatine synthesis in the body can also be increased by ingestion of creatine present in meat (eg, 5-10 milligrams / kilogram body weight / day for an average carnivorous person and nearly zero for vegetarian diets).

  The accumulation of hydronium ions formed during glycolysis and lactic acid accumulation (anaerobic metabolism) during sustained intense exercise or exercise sustained under local hypoxic conditions The internal pH can be greatly reduced. Reduced pH can compromise the function of the creatine-phosphoryl creatine system. A decrease in intracellular pH can affect other functions in the cell, such as the function of contractile proteins in muscle fibers.

  Dipeptides of β-alanine and histidine, including carnosine (β-alanyl-L-histidine), anserine (β-alanyl-L-1-methylhistidine), or valenine (β-alanyl-L-3-methylhistidine) (Also referred to herein as peptides), and their methylated analogs, are present in muscle of humans and other vertebrates. Carnosine is found in considerable amounts in, for example, human and horse muscles. Anserine and carnosine are found, for example, in the muscles of dogs, camels and many birds. Anserine is the major β-alanyl histidine dipeptide in many fish. Valenin is the major β-alanyl histidine dipeptide in several species of aquatic mammals and reptiles. In humans, horses, and camels, the highest concentration of β-alanyl histidine dipeptide is found in glycolytic muscle fibers (types IIA and IIB) that are widely used during intense exercise. Lower concentrations are found in muscle fibers with slow contraction in the oxidized form (type I). For example, see (Non-Patent Document 1). Carnosine is known to contribute to hydronium ion buffering capacity in different muscle fiber types and up to 50% of the total in horse type II fibers.

Dunnett, M. & Harris, R.C.Equine Vet. J., Suppl. 18, 214-217 (1995)

  The present invention seeks to provide methods and compositions for enhancing anaerobic work capacity in tissues.

  The present invention is a method for enhancing anaerobic working capacity in a tissue, comprising the following steps: (a) β-alanyl histidine dipeptide and glycine, insulin, insulin mimetic, or insulin action modifier And (b) adding β-alanine and at least one of glycine, an insulin mimetic, or an insulin action modifier to the tissue in an amount effective to increase β-alanyl histidine dipeptide synthesis in the tissue. Administering, thereby increasing the anaerobic working capacity in the tissue. The present invention provides a method for regulating the concentration of hydronium ions in a tissue, comprising the following steps: (a) β-alanyl histidine dipeptide and glycine, insulin, insulin mimetic, or insulin action modifier And (b) administering β-alanine and at least one of glycine, an insulin mimetic, or an insulin action modifier to the tissue in an amount effective to increase hydronium ion concentration in the tissue. Wherein the method comprises:

  In one embodiment of the method, administering β-alanine and at least one of glycine, an insulin mimetic, or an insulin action modifier to the tissue comprises oral administration, administration to blood or plasma, or combinations thereof including. The β-alanyl histidine dipeptide can include carnosine, anserine, or valenin, or analogs or mimetics thereof.

  The present invention relates to an amino acid selected from the group consisting of a peptide comprising glycine, insulin, an insulin mimetic, or an insulin action modifier and β-alanine, a chemical derivative of β-alanine, and β-alanine or an analog thereof. Alternatively, a composition comprising a mixture with a composition having an active derivative thereof is provided. In one embodiment, β-alanine comprises β-alanyl histidine dipeptides such as carnosine, anserine, or valenin, or analogs thereof. The composition can further comprise at least creatine or a carbohydrate.

  In one embodiment, the insulin mimetic is D-pinitol (3-O-methyl-thyleusitol), 4-hydroxyisoleucine, demethyl-asteriquinone B-1 compound, alpha lipoic acid, R-alpha lipoic acid, guanidinopropionic acid , Vanadium compounds, vanadium complexes, or synthetic phosphoinositol glycan peptides. The insulin action modifier can be a sulfonylurea, thiazolidinedione, or biguanide.

  In another embodiment, the composition is a pharmaceutical composition, a dietary supplement, or a sports drink. The dietary supplement or sports drink can be a human supplement. The pharmaceutical composition can be formulated for human use.

  The present invention is at least 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, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0 6.5 grams or more of a composition comprising a peptide or ester having β-alanine or an analog or mimetic thereof. The present invention provides at least about 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, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6. Of an injectable form comprising 0, 6.1, 6.2, 6.3, 6.4, 6.5 grams or more of a peptide or ester having β-alanine (or an analog or mimetic thereof) A composition is provided. In one embodiment, the peptide comprises a β-alanyl histidine dipeptide, such as carnosine, anserine, or valenin, or an analog or mimetic thereof.

  The present invention is at least 200, 225, 250, 275, 300, 325, 350, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 750, 775, 800. , 825, 850, 875, 900, 925, 950, 975 or 1000 mg or more of β-alanine or β-alanine analogs or mimetics are provided. In one embodiment, the composition is formulated for ingestion or is in an injectable form. Ingestible dosage forms can be beverages, gels, foods, or tablets. Peptides can include β-alanyl histidine such as carnosine, anserine or valenin, or analogs or mimetics thereof.

  The present invention is a method for increasing the anaerobic working capacity of a tissue in a subject comprising the following steps: (a) (i) glycine, insulin, an insulin mimetic, or an insulin action modifier, β-alanine, β A mixture of a chemical derivative of alanine and a composition having an amino acid selected from the group consisting of peptides comprising β-alanine or analogs or mimetics thereof, or an active derivative thereof, (ii) at least in injectable form Providing a composition comprising a peptide or ester containing 0.5 g β-alanine, or (iii) at least 200 mg β-alanine, and (b) effective to enhance the anaerobic capacity of the tissue Administering the composition to the subject in a suitable amount. In one embodiment, the total 24 hour dose of β-alanine is at least about 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, 2.0, 2.5, 3.0, 3.5, 4.0, 4. 5, 5.0, 5.5, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5 grams or more. The total 24-hour dose of β-alanine is between about 0.2 g to about 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0 grams or more It can be. The composition may be given over a period of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 days or more. The composition may be given over a period of at least about 3 days to about 2, 3, 4 weeks or longer. β-alanine can include β-alanyl histidine dipeptides such as carnosine, anserine, or valenin, or analogs or mimetics thereof. The total dose of β-alanyl histidine dipeptide over 24 hours is at least about 0.5 g, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6. Can be 1, 6.2, 6.3, 6.4, 6.5 grams or more. The total dose of β-alanyl histidine dipeptide over 24 hours is about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 grams or more Can be larger. The total dose of β-alanyl histidine dipeptide over 24 hours can be from about 5 g to about 16 g. The composition can be administered in multiple doses. The composition may be administered at least 2 to 8 times over a 24 hour period. In one embodiment, about 200 mg, 225, 250, 275, 300, 325, 350, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975 or 1000 mg of β-alanine (or an analog or mimetic thereof), and / or about 500 mg (or about 200 mg, 225, 250, 275, 300, 325, 350, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, Or 1000 mg of carnosine (or an analogue or倣体) is 8 or more times from about twice a day for a period of weeks (e.g., 2,3,4,5,6,7,8 or more times per day) is administered. In one embodiment, at least about 2 g of β-alanine, or at least about 5 g of carnosine is administered from about 2 to 8 times per day over a period of about 2, 3 or 4 days.

  In one embodiment, the amount of the composition of the invention administered is increased daily. The amount of the composition of the invention administered can be increased weekly. The composition can be administered for a treatment period lasting at least about 4 weeks.

  Although the present invention is not limited to any particular mechanism of action, the present invention is a method for adjusting the concentration of hydronium ions in tissue in a subject comprising the following steps: (a) (i) An amino acid selected from the group consisting of glycine, insulin, an insulin mimetic, or an insulin action modifier and β-alanine, a chemical derivative of β-alanine, and a peptide comprising β-alanine or an analog or mimetic thereof, or A composition comprising a mixture with a composition having an active derivative thereof, (ii) an injectable form of a peptide or ester containing at least 0.5 grams of β-alanine, or (iii) at least 200 mg of β-alanine. And (b) applying the composition to the subject in an amount effective to adjust the hydronium ion concentration in the tissue. Administering the method.

  In one aspect, the invention features methods and compositions for enhancing anaerobic work capacity of muscle and other tissues. The methods and compositions of the present invention provide for the simultaneous accumulation of creatine and / or β-alanyl histidine dipeptides, or β-alanine and L-histidine analogs within tissues in the body. The method includes ingesting or injecting the composition into the body. In one embodiment, the composition is a mixture of a compound capable of increasing creatine availability and uptake and a precursor for synthesis and accumulation of β-alanyl histidine dipeptide in human and animal tissues. The composition of the present invention can induce the synthesis and accumulation of β-alanyl histidine dipeptide in the human or animal body when introduced into the body.

  This composition comprises chemical derivatives and analogs of β-alanine such as β-alanine, esters of β-alanine, peptides of β-alanine, such as β-alanine, carnosine, anserine, and valenin, and analogs thereof. Can be included. The composition may also contain L-histidine and mixtures thereof. Each β-alanine and / or L-histidine is formulated or administered as an individual amino acid, or as a component of a dipeptide (eg, carnosine, anserine, and / or valenin), oligopeptide, or polypeptide. Can do. β-alanine, L-histidine, carnosine, anserine, and / or valenin, or peptides of β-alanine can be active derivatives. An active derivative is derived from or is a precursor of a substance and functions in the same or similar state as the substance in the body or is processed into a substance when placed in the body It is. Examples include, for example, esters and amides. The composition can also include any one or more of creatine, carbohydrates, insulin, insulin mimetics, insulin action modifiers or glycine. The compositions of the present invention can be used in the preparation of dietary supplements (eg, including beverages, gels, foods) or pharmaceutical compositions for humans or animals. The composition of the present invention can be used in any of the methods of the present invention.

  In one aspect, the invention features a composition and method for adjusting hydronium ion concentration in a tissue. The method comprises the steps of providing the blood or plasma with an amount of β-alanine effective to increase β-alanylhistidine dipeptide synthesis in the tissue, thereby contacting the tissue with the blood or plasma, thereby Increasing the concentration of β-alanyl histidine. β-alanyl histidine can be carnosine, anserine, or valenin. The method can include donating an amount of L-histidine to blood or plasma effective to increase β-alanyl histidine dipeptide synthesis.

  In one aspect, the invention features a method for increasing the anaerobic capacity of a tissue. The method includes providing an effective amount of β-alanine to the blood or plasma to increase β-alanyl histidine dipeptide synthesis in the tissue and increasing β-alanyl histidine dipeptide synthesis in the tissue. Providing an effective amount of L-histidine to the blood or plasma and contacting the tissue with the blood or plasma. The concentration of β-alanyl histidine increases in the tissue.

  In another aspect, the method can include increasing the concentration of creatine in the tissue. The increasing step can include donating an amount of creatine to the blood or plasma that is effective to increase the concentration of creatine in the tissue (eg, by donating creatine to the blood or plasma). .

  The donating step of this method is a β-alanine or a β-alanine peptide such as carnosine, anserine, and valenin that is hydrolyzed to their constituent amino acids upon ingestion and is a source of β-alanine for the body Ingestion, infusion (eg, injection), or a combination of ingestion and infusion. The methods of the invention also include donating L-histidine, creatine, carbohydrates, insulin, insulin mimetics, insulin action modifiers and / or glycine.

  In yet another aspect, the method can include increasing the concentration of insulin in the blood or plasma. The concentration of insulin can be increased, for example, by injection of insulin. The methods of the invention can also include injection, ingestion, or other forms of insulin mimic delivery to the body (also referred to as a subject) known to those skilled in the art. Examples of insulin mimetics include, but are not limited to, D-pinitol (3-O-methyl-tyrosinositol), 4-hydroxyisoleucine, L783,281 (demethyl-asteriquinone B-1 compound), alpha lipo Examples include acids, vanadium compounds such as R-α lipoic acid, guanidinopropionic acid, vanadyl sulfate, or vanadium complexes such as peroxovanadium, and synthetic phosphoinositol glycans (PIG peptides). In addition or alternatively, the methods of the invention can include the use of an insulin action modifier to enhance or inhibit the action of insulin in the body. Examples of insulin action modifiers can include, but are not limited to, sulfonylureas, thiazolidinediones, and biguanides.

  In yet another embodiment, the method includes donating glycine to the body. Glycine is believed to be able to suppress the release of blood glucose in the blood after meal intake. Glycine may increase insulin sensitivity by promoting the uptake of more glucose. Thus, the methods include donating glycine alone or with insulin, insulin mimetics, or insulin action modifiers in the compositions and methods of the invention. Glycine can be provided in various forms, for example, alone or with other substances such as in a dietary supplement. Alternatively, glycine can be obtained from other sources such as gelatin.

  The tissue referred to in the present invention may be skeletal muscle.

  In one embodiment, the present invention provides a composition for performing the method of the present invention. Accordingly, one aspect of the present invention is a β-alanine, β-alanyl histidine peptide (or analog or derivative thereof), creatine, insulin, insulin mimetic, or insulin action modification for carrying out the methods of the present invention. Contemplated are compositions having one or more active ingredients including factors, glycine, and carbohydrates. The present invention further contemplates the use of complex compositions formulated to donate one or more active ingredients to the body for the purpose of performing the methods of the present invention.

  Thus, in an exemplary embodiment, the present invention consists essentially of β-alanine or a peptide source of β-alanine, from about 39% to about 99% by weight carbohydrate, and up to about 60% by weight water. It is a composition. The composition can comprise from about 0.1% to about 20% by weight of β-alanine (in free or bound form). The composition can comprise from about 0.1% to about 20% by weight of L-histidine.

  The carbohydrate can be a simple carbohydrate (eg, glucose).

  In another aspect, the invention consists essentially of a composition consisting of β-alanine or a peptide source of β-alanine, from about 1 wt% to about 98 wt% creatine source, and up to about 97 wt% water. It is characterized by being. The composition comprises from about 0.1% to about 98% β-alanine by weight. The peptide source can include L-histidine, and the composition can include from about 0.1% to about 98% by weight L-histidine from the source.

  The peptide source can be an amino acid, dipeptide, oligopeptide, polypeptide, or a mixture of active derivatives thereof.

  The composition can be a dietary supplement. The creatine source can be creatine monohydrate.

  The concentration of a component in the blood or plasma, including β-alanine, can be increased by infusion (ie, injection) or ingestion of an agent that can function to cause an increase in plasma concentration. The composition can be taken at a dose of about 100 milligrams to about 800 grams or more per day. The dose can be administered once or multiple times daily.

  Increased creatine and β-alanyl histidine dipeptides in muscles increase cellular tolerance to increased hydronium ion production by anaerobic work, and result in increased endurance during exercise before fatigue develops Can do. The present compositions and methods can contribute to correcting the loss of β-alanine, L-histidine, or creatine due to degradation or leaching of these components during food preparation or processing. The present compositions and methods can also contribute to correcting these component deficiencies due to vegetarian diet.

  The methods and compositions are used to increase β-alanyl histidine dipeptides in sportsmen, athletes, bodybuilders, synchronized swimming athletes, soldiers, the elderly, racehorses, working and racing dogs, and hunting birds. Can avoid or delay the onset of muscle fatigue.

FIG. 1 shows that for 30 days before feeding β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, 3 times per day, respectively) and every 2 hours after feeding. It is a graph showing the change of the density | concentration of (beta) -alanine in the plasma of five horses. FIG. 2 shows that β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, 3 times per day, respectively) for 30 days before feeding and every 2 hours after feeding. It is a graph showing the change of the density | concentration of L-histidine in the plasma of five horses. FIG. 3a shows, as detailed above, the change in the concentration of β-alanine in the plasma of six horses before feeding and every hour after feeding β-alanine and L-histidine. It is a graph showing the contrast in. FIG. 3b shows, as detailed above, the change in the concentration of β-alanine in the plasma of six horses before feeding and every hour after feeding β-alanine and L-histidine. It is a graph showing the contrast in. FIG. 3c shows, as detailed above, the change in the concentration of β-alanine in the plasma of six horses before feeding and every hour after feeding β-alanine and L-histidine. It is a graph showing the contrast in. FIG. 3d shows, as detailed above, the change in the concentration of β-alanine in the plasma of 6 horses before and after feeding β-alanine and L-histidine. It is a graph showing the contrast in. FIG. 3e shows, as detailed above, the change in the concentration of β-alanine in the plasma of 6 horses before feeding and every hour after feeding β-alanine and L-histidine. It is a graph showing the contrast in. FIG. 3f shows the change in the concentration of β-alanine in the plasma of 6 horses before and after feeding β-alanine and L-histidine, as detailed above. It is a graph showing the contrast in. FIG. 4a shows the change in the concentration of L-histidine in the plasma of 6 horses before and after feeding β-alanine and L-histidine, as detailed above. It is a graph showing the contrast in. FIG. 4b shows, as detailed above, the change in the concentration of L-histidine in the plasma of 6 horses before feeding and every hour after feeding β-alanine and L-histidine. It is a graph showing the contrast in. FIG. 4c shows the change in the concentration of L-histidine in the plasma of 6 horses before and after feeding with β-alanine and L-histidine, as detailed above. It is a graph showing the contrast in. FIG. 4d shows the change in the concentration of L-histidine in the plasma of 6 horses before and after feeding β-alanine and L-histidine, as detailed above. It is a graph showing the contrast in. FIG. 4e shows, as detailed above, the change in the concentration of L-histidine in the plasma of 6 horses before feeding and every hour after feeding β-alanine and L-histidine. It is a graph showing the contrast in. FIG. 4f shows the change in the concentration of L-histidine in the plasma of 6 horses before and after feeding the β-alanine and L-histidine feed, as detailed above. It is a graph showing the contrast in. FIG. 5 shows, as detailed above, the average concentration of β-alanine in horse plasma (n = 6) before and every hour after feeding β-alanine and L-histidine. It is a graph showing the contrast in the change of. FIG. 6 provides a feed of β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, 3 times per day, respectively) on the first and last day of the 30-day feeding period. FIG. 5 is a graph showing the contrast in the change in mean concentration of L-histidine in horse plasma (n = 6) before and every hour after feeding. FIG. 7 shows the increase in carnosine concentration in type II skeletal muscle fibers (average of type IIA and type IIB fibers) in 6 thoroughbred horses and the day between supplementation from day 1 to day 30 FIG. 6 is a graph showing the correlation with the increase in the area under the plasma β-alanine concentration-time curve (AUC _{(0-12 hr)} ) over the first 12 hours. FIG. 8 is a graph showing the average results of administration of β-alanine, broth, or carnosine to subjects. FIG. 9 is a graph representing the mean change in plasma β-alanine over 9 hours of treatment. FIG. 10 is a graph showing the mean change in plasma β-alanine over 9 hours after ingestion of 10 mg / kg body weight of β-alanine. FIG. 11 is a graph depicting the mean (n = 6) plasma β-alanine concentration over 24 hours on days 1 and 30 of the treatment period. FIG. 12 is a graph showing changes in muscle carnosine concentration before and after treatment in different subjects. Red circles indicate muscle concentration before replenishment. FIG. 13 is a graph depicting carnosine muscle concentration (mean + SD) before and after supplementation in three different treatment groups. FIG. 14 is a graph depicting histidine muscle concentration (mean + SD) before and after supplementation in three different treatment groups. FIG. 15 is a graph illustrating data showing muscle concentration (average + SD) of taurine before and after supplementation in four different treatment groups. FIG. 16 is a graph for explaining data indicating taurine muscle concentration (average + SD) before and after supplementation in different subjects. FIG. 17 illustrates a table of data described in detail below as Table 9.

  The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other advantages and features of the invention will be apparent to those skilled in the art from the detailed description, drawings, and claims.

  All publications, patents and patent applications cited herein are hereby expressly incorporated by reference.

  Like reference symbols in the various drawings indicate like elements.

(Detailed explanation)
The present invention relates to compositions comprising β-alanine, peptides of β-alanine, analogs and derivatives thereof, β-alanyl histidine dipeptides (eg, carnosine, anserine, and valenin), and tissue anoxic work capacity. A method of using these compositions to enhance blood pressure, comprising providing blood or plasma with an amount of β-alanine effective to increase β-alanylhistidine dipeptide synthesis in tissue. provide. The β-alanyl histidine dipeptide can include β-alanine peptides such as carnosine, anserine, and valenin. In one embodiment, they can have a pKa value between about 6.8 and 7.1. In one embodiment, they may be involved in the regulation of intracellular pH homeostasis while muscle contraction and fatigue progress. The content of other substances involved in hydronium ion buffering, such as amino acid residues in proteins, inorganic and organic phosphates and bicarbonates, can be constrained by their involvement in other cellular functions. In one embodiment, β-alanyl histidine dipeptide provides an effective means of accumulating pH sensitive histidine residues in cells. Variation in muscle β-alanyl histidine dipeptide concentration affects the anaerobic performance of individual athletes.

  β-alanyl histidine dipeptide is synthesized in the body from β-alanine and L-histidine. These precursors can be produced in the body or made available through food, including by ingestion of ingested β-alanyl histidine dipeptides. In the body, β-alanine is transported to tissues such as muscle. Under typical dietary conditions, the concentration of β-alanine in human and horse plasma is low compared to the concentration of L-histidine. These concentrations should be considered in relation to the affinity of carnosine synthase, carnosine synthetase for the substance, as determined by the Michaelis-Menten constant (Km). The Km for histidine is about 16.8 μM. The Km for β-alanine is between about 1000 and 2300 μM. The low affinity of carnosine synthetase for β-alanine and the low concentration of β-alanine in muscle makes the concentration of β-alanine in muscle rate-limiting for the synthesis of β-alanyl histidine dipeptides. Explain that.

  Increasing the amount of β-alanyl histidine dipeptide in muscle has a positive effect on muscle performance and the amount of work that can be done by the muscle. Therefore, it is desirable to increase the synthesis and accumulation of β-alanyl histidine dipeptide in tissues in the human or animal body.

  Synthesis and accumulation of β-alanyl histidine dipeptide in the human or animal body increases creatine in the body, increases blood or plasma concentration of β-alanine, increases blood or plasma concentration of β-alanine and creatine Or by increasing blood or plasma concentrations of β-alanine, L-histidine, and creatine. The increase in dipeptide can occur simultaneously with the increase in β-alanine concentration.

  In one embodiment, the compositions and methods of the present invention comprise β-alanine, a peptide of β-alanine, L-histidine, creatine, carnosine, anserine, and / or valenin, and / or an active derivative or analog thereof alone. Alternatively, it can be used to increase plasma concentrations of β-alanine, L-histidine, and / or creatine by ingestion or infusion in various combinations. The composition of the present invention can be administered orally, enterally, or parenterally. For example, the compositions of the present invention can be taken orally or injected through the skin with a paint or patch.

  The composition can include a carbohydrate (eg, a simple carbohydrate), insulin, or a substance that promotes the production of insulin. The composition can also include glycine, insulin, insulin mimetics, and / or insulin action modifiers.

  The composition can be a dietary supplement that can be ingested, injected, or absorbed through the skin. Preferably, the composition can be administered in one or more doses per day. The dose of β-alanine can be between about 1 milligram to about 200 milligrams / kilogram body weight, or the dose of β-alanine peptide (eg, carnosine) is from 2.5 milligrams to 500 milligrams / kilogram body weight. be able to. In one embodiment, the total amount of β-alanine (or other composition of the invention) administered can be at least 200 mg, 200 mg to 5 g, or 5 g or more per day for humans. A single dose of the active ingredient, eg, β-alanine, carnosine, anserine, or valenin, or a mixture thereof can be formulated to be an amount of about 200, 400, 800 mg, or more. The dose of creatine (eg, creatine monohydrate) or other compositions of the invention can be between about 5 milligrams and 200 milligrams / kilogram body weight. The dose of L-histidine, or other composition of the present invention, can be between about 1 milligram and 100 milligrams / kilogram body weight. A single carbohydrate (eg, glucose) dose, or a dose of the composition of the invention can be between about 0.5 and 2.0 grams / kilogram body weight.

  For an 80 kilogram person, the appropriate daily dose is 0.08 to 16.0 grams of β-alanine, or 200 to 40 grams of β-alanine peptide, and 0.4 grams of creatine monohydrate. To 16.0 grams, L-histidine from 0.08 grams to 8.0 grams, or glucose or other simple carbohydrates from 40 grams to 160 grams. The composition can be in the form of a solid, liquid, or suspension that can be ingested or injected into the body. The composition can be taken by a human in an amount between 0.08 and 1000 grams per day or more and can be taken in one or more doses throughout the day. In animals, daily intake will be adjusted by body weight.

  In one embodiment, the total dose of a peptide of β-alanine that can be administered per day, such as carnosine, anserine, or valenin is at least 500 mg, between about 500 mg to about 5 g, between about 5 g to about 16 g, Or more than 16g. A single dose of a peptide of β-alanine, creatine, anserine, or valenin, or a mixture thereof includes an amount of 0.5, 1, 1.5, or 2 g (for each or all or a mixture) It can be formulated so as to be about the peptide in the formulation.

  For humans and animals, the composition may be as follows.

(A) 1% to 99% by weight β-alanine or 1% to 99% by weight β-alanine peptide;
1% to 99% by weight of creatine monohydrate; and 0% to 98% by weight of water;
(B) 1% to 98% β-alanine or 1% to 98% β-alanine peptide;
1% to 98% by weight of L-histidine;
1% to 98% by weight of creatine monohydrate; and 0% to 97% by weight of water;
(C) 1% to 20% by weight β-alanine or 1% to 20% by weight β-alanine peptide;
39% to 99% glucose or other simple carbohydrate; and 0% to 60% water; or (d) 1% to 20% β-alanine or 1% to 20% by weight. a peptide of β-alanine;
1% to 20% by weight of L-histidine;
39% to 99% glucose or other simple carbohydrate; and 0% to 60% water.

  In one embodiment, the composition is applied to the body for a period of at least 3 days, 3 days to 2 weeks, 2 weeks to 4 weeks, or longer. In certain dosing regimes, the daily dose is gradually increased or decreased. This can be done daily, every 2-3 days, or weekly.

  Specific examples of methods and compositions for increasing the anaerobic working capacity of muscles and other tissues are given below.

Example 1
The effect of supplementing a normal diet containing multiple daily doses of β-alanine and L-histidine on carnosine concentrations in type I, IIA, and IIB skeletal muscle fibers of thoroughbred horses was evaluated. Six experimental thoroughbred horses (three female foals and three steers), 4-6 years of age, are fed for 1 month (30 days) prior to the start of the supplementation period Received adjustment (pre-supply period). During the dietary adjustment period, each horse receives three doses per day (at 8:30, 12:30, and 16:30 respectively), 1 kilogram of pelleted feed (Spillers racehorse cubes) and complex and simple carbohydrates. Feed was provided with 1 kilogram of soaked beet pulp as a source. Soaked hay (3 kg dry weight) was also given twice daily (9 and 17:00). Water was given accordingly.

  The same feeding method was carried out during the supplementation period. However, each hard feed flour was supplemented with β-alanine and L-histidine (free base). β-alanine and L-histidine were mixed directly into normal feed. Individual doses of β-alanine and L-histidine were calculated according to body weight. β-alanine was administered at 100 milligrams / kilogram body weight and L-histidine was administered at 12.5 milligrams / kilogram body weight. Feeding began on day 1 of the protocol and was interrupted at the end of day 30. Heparinized blood samples (5 ml) were taken on days 1, 6, 18, 24, and 30. On day 1 and day 30, blood samples were taken on each day before the first feeding and every hour for a total of 12 hours. During the three sampling days in between, blood was collected before the first feeding and 2 hours after each subsequent feeding. Muscle biopsy from each horse's right gluteus muscle (m. Gluteus medius) using the Bergstrom-Stille percutaneous biopsy needle after local skin anesthesia on the day before the start of supplementation (day 0) Went. Subsequent muscle biopsies were taken as close as possible to the first sampling site immediately after the replenishment period (day 31) was over. Horse clinical monitoring was performed daily. This includes visual inspection and body weight measurement, twice daily measurement of rectal temperature, and weekly blood sampling for clinical biochemistry and hematology. During the course of the study, the horse did not receive any formal training or exercise, but allowed one hour of free movement every day.

Individual muscle fiber fragments cut from a lyophilized muscle biopsy were preincubated at pH 4.5 by modification of the method described in Kaiser and Brook, Arch. Neurol. , 23: 369-379 (1970) . It was subsequently characterized as either type I, type IIA, or type IIB by histochemical staining for myosin ATPase activity at pH 9.6.

Heparinized plasma samples were extracted by high performance liquid chromatography (HPLC) and analyzed for β-alanine and L-histidine concentrations. Individually weighed muscle fibers are described in Dunnett and Harris, "High-performance liquid chromatographic determination of imidazole dipeptides, histidine, 1-methylhistidine and 3-methylhistidine in muscle and individual muscle fibers," J. Chromatogr. B. Biomed Extracted by HPLC and analyzed for carnosine according to the method described in Appl. , 688: 47-55 (1997).

  Using one-way analysis of variance (ANOVA), differences in carnosine concentrations in the fiber type before and after supplementation were confirmed in horses. In cases where differences were detected, significance was determined using multiple comparison tests (Fisher's PLSD method).

  There was no preference problem with adding β-alanine and L-histidine to the feed. During the 30 days of supplementation, no physiological or behavioral adverse effects from supplemented diet were observed in any of the horses. Significant changes in body weight were not recorded, and rectal temperature remained within the normal range. No clinical biochemical or hematological acute or chronic changes were observed. No β-alanine was detected in any plasma of horses before the start of supplementation. The lower limit of quantification by the assay used for β-alanine in plasma was 3 micromolar (μM). Plasma L-histidine concentrations in 6 horses before the start of supplementation were between 36.6 and 54.4 μM.

  The individual changes in plasma β-alanine and L-histidine concentrations for 5 of 6 horses over all sampling days are shown in FIGS. 1 and 2, respectively. There was a tendency for plasma β-alanine and L-histidine concentrations before feeding to increase as the supplementation period increased. Furthermore, the plasma concentration response to supplementation also increased over the 30-day supplementation period. The reaction was greater for β-alanine.

A comparison of the changes in plasma β-alanine and L-histidine concentrations for the six individual horses between the first and last day of the supplementation period before feeding the first feed of the day and every hour thereafter is shown in FIG. And 3b and FIGS. 4a and 4b, respectively. Figures 3a, 3b, 3c, 3d, 3e, and 3f show β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg respectively) on the first and last days of the 30 day period of feed supplementation. It is a graph showing the contrast in the change of the density | concentration of (beta) -alanine in the plasma of 6 horses before giving the feed of a kilogram body weight (3 times per day) and every 1 hour after giving. Figures 4a, 4b, 4c, 4d, 4e, and 4f show β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg respectively) on the first and last days of the 30 day period of feed supplementation. It is a graph showing the contrast in the change of the L-histidine density | concentration in the plasma of 6 horses before giving the feed of a kilogram body weight (3 times per day) and every 1 hour after giving. FIG. 5 shows the feed of β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, 3 times per day, respectively) on the first and last day of the 30 day period of feed supplementation. It is a graph showing the contrast in the change of the average density | concentration of (beta) -alanine in the horse plasma (n = 6) before giving and every other hour after giving. The mean (SD) change in plasma β-alanine concentration (n = 6) over 24 hours on the first day (day 1) and last day (day 30) of the supplementation period is compared in FIG. The mean plasma β-alanine concentration over 24 hours area under the time curve (AUC _{(0-24 hr)} ) was very large at day 30 of supplementation.

The mean (SD) change (n = 6) in plasma L-histidine concentration over 24 hours on the first day (day 1) and last day (day 30) of the supplement period is compared in FIG. The area under the mean plasma β-alanine concentration versus time curve over 24 hours (AUC _{(0-24 hr)} ) was greater at day 30 of supplementation. A larger AUC for plasma β-alanine on the last day of supplementation (day 30) compared to the first day of supplementation (day 1) indicates that β-alanine uptake from the horse's gastrointestinal tract as the supplementation proceeds. Suggests an increase. Similar effects were observed for changes in plasma L-histidine concentration during the supplementation period. In each case, the highest plasma concentrations of β-alanine and L-histidine were produced about 1-2 hours after feeding.

A total of 397 individual skeletal muscle fibers (192 before supplement, 205 after supplement) were cut from 6 horses and analyzed for carnosine. Mean (SD) carnosine concentration expressed in millimoles / kilogram dry weight (mmol · kg ⁻¹ dW) in type I, type IIA, and type IIB skeletal muscle fibers before and after supplementation from six individual horses Is given in Table 1. N in Table 1 is the number of individual muscle fibers analyzed. As a result of 30 days supplemented with β-alanine and L-histidine, the average carnosine concentration was increased in type IIA and type IIB fibers of all six horses. These increases were statistically significant in 7 cases. The increase in mean carnosine concentration in type IIB fibroskeletal muscle fibers was statistically significant in 5 out of 6 horses. The increase in mean carnosine concentration in type IIA skeletal muscle fibers was statistically significant in 2 of 6 horses.

Table 2 lists the absolute increase (eg, mmol kg ⁻¹ dW) and increase in mean carnosine concentration in type IIA and IIB skeletal muscle fibers from six horses.

It was observed that individual horses that showed a large increase in muscle carnosine concentration after 30 days of supplementation also exhibited a large increase in plasma β-alanine AUC between days 1 and 30 of the supplementation period. According to FIG. 7, a significant correlation (r = 0.986, p <0.005) for 5 of the 6 horses showed an increase in mean carnosine concentration averaged over type IIA and type IIB skeletal muscle fibers. , _Observed during the first 12 hours of supplementation during the first 12 hours with an increase in plasma β-alanine AUC (AUC _{(0-12 hr)} ). Only 5 horses were used to calculate the regression line. Horse 6 (filled circles) did not show a noticeable increase in plasma β-alanine concentration that was greater than the increase observed on day 1 until the last day of supplementation. This was different from the other five horses that progressively increased with each sampling day. For this reason, Horse 6 was excluded from the calculation of the regression equation.

An increase in muscle carnosine concentration as a result of 30-day supplementation with β-alanine and L-histidine will result in a direct increase in total muscle buffer capacity. This increase can be calculated using the Henderson-Hasselbalch equation. Table 3 shows the calculated values for increased muscle buffering capacity of type IIA and type IIB skeletal muscle fibers in six Thoroughbred horses.

(Example 2)
The effect of supplementing a normal diet, including daily and multiple doses of β-alanine in free or peptide-bound state, on β-alanine and β-alanyl dipeptide concentrations in human plasma was evaluated. The plasma concentration of β-alanine in 6 normal subjects was observed after consuming broth supplying approximately 40 milligrams / kilogram body weight of β-alanine. Dose of 10 and 20 milligrams / kilogram body weight of β-alanine was also given.

  Bros prepared as follows: Fresh chicken breasts (without skin and bone removed) were minced and boiled for 15 minutes with water (1 liter for every 1.5 kg of chicken). The remaining chicken was crushed and removed. Seasoned carrots, onions, celery, salt, pepper, basil, parsley and tomato puree and reboiled for another 15 minutes, then cooled to 4 ° C. before final rinsing through fine muslin. Yield from 1.5 kilograms of chicken and 1 liter of water was 870 mL broth. A portion of the broth was measured for total β-alanyl-dipeptide content (eg, carnosine and anserine) and β-alanine. Typical analytical values were as follows:

Total β-alanyl-dipeptide 74.5 mM
Free β-alanine 5.7 mM
Six male subjects were in normal health and were 25-53 years old as shown in Table 4. The investigation began after an overnight fast (eg, a minimum of 12 hours after the last meal containing meat). Prior to the start of the study, subjects were given the option of taking a small amount of hot water. The catheter insertion began at 8:30 and the study began at 9 o'clock.

  As a control, 8 ml / kg body weight of water was ingested (for example, 600 mL for a 75 kg body weight subject).

  In one session, 8 milliliters / kilogram body weight broth containing about 40 milligrams / kilogram body weight of β-alanine (eg, in the form of anserine and carnosine) was ingested. This resulted in an intake of 600 milliliters of broth containing a total of 3 grams of β-alanine for a subject weighing 75 kilograms. In another session, an additional 3 milliliters / kilogram body weight of fluid containing a test amount of β-alanine along with 5 milliliters / kilogram body weight of water was ingested. In all sessions, subjects received an additional 8 milliliters / kilogram body weight of water (in a 50 mL volume) between 1-2 hours after ingestion. Six hours later I gave vegetarian pizza. Subsequently, a normal meal was made 8 hours later.

A 2.5 milliliter venous blood sample was taken through the indwelling catheter every 10 minutes for the first 90 minutes and then 120, 180, 240 and 360 minutes. The blood sample was dispensed into a tube containing lithium-heparin as an anticoagulant. The catheter was maintained by rinsing with saline. Plasma samples were analyzed by HPLC according to the method described in Jones & Gilligan (1983) J. Chromatogr. 266: 471-482 (1983).

The treatment assignments under investigation of β-alanine absorption are summarized in Table 4. The estimated equivalent dose of β-alanine is shown in Table 3.

  The plasma concentration curve after each treatment is represented in the graph of FIG. Table 4 summarizes the mean results of treatment with β-alanine, broth, or carnosine. Plasma β-alanine was below the detection limit in all subjects on control treatment. Neither carnosine nor anserine was detected in plasma after ingestion of chicken broth or other treatments. Ingestion of broth resulted in the highest concentration of 427.9 (SD 161.8) μM in plasma. Administration of carnosine corresponding to 20 milligrams / kilogram body weight of β-alanine in one subject resulted in an equivalent increase in plasma β-alanine concentration.

  All treatments except controls increased plasma taurine levels. The change in taurine concentration strictly reflected the change in β-alanine concentration. Administration of broth, a natural food, results in an equivalent increase in plasma taurine, which indicates that the response usually occurs after most meals.

(Example 3)
The effect of administering three doses of 10 mg / kg body weight of β-alanine per day for 7 days (ie, administered in the morning, noon, and night) on the plasma concentration profile of β-alanine and taurine was investigated. Plasma concentration profiles after administration of 10 milligrams / kilogram body weight of β-alanine were investigated at the beginning and end of the 7 day period in 3 subjects. Meanwhile, subjects were given 3 doses of β-alanine per day.

Three male subjects aged 33-53 in normal health were examined. Subjects received 3 doses of 10 mg / kg body weight of β-alanine per day for 8 days. Two subjects were then given 3 doses of 20 milligrams / kilogram body weight per day for an additional 7 days (days 9-15). Subjects reported to the blood collection lab on day 1 (before any treatment) and on days 8 and 15 at 8 am after an overnight fast. Subjects were asked not to eat any meaty meals for 12 hours prior to the study. On each of these 3 day test days, the subject was inserted with a catheter and an initial blood sample was taken when β-alanine was administered at 9 am, 12 noon, and 3 pm or around that time. . Blood samples were taken after 30, 60, 120 and 180 minutes and analyzed for changes in plasma concentrations of β-alanine and taurine. A 24-hour urine sample was collected over each study day and analyzed by HPLC to determine β-alanine and taurine excretion. The treatments are summarized in Table 5.

  Plasma β-alanine concentrations are summarized in FIG. Each time, a peak of β-alanine concentration occurred 30 minutes or 1 hour after ingestion, and dropped to a basal level of 0 to 10 μmol in 3 hours immediately before the next administration. The responsiveness of the 8th day of treatment tended to be lower than the 1st day as shown by the area under the plasma concentration curve.

Example 4
Three doses of β-alanine at 40 mg / kg body weight per day for two weeks (ie morning, noon, and night) for muscle carnosine content and isometric endurance at 66% of maximum voluntary contractile force Was examined.

Six normal male subjects aged 25 to 32 years who had no signs of metabolic or muscle disease were recruited and investigated. Subjects were asked about recent diet and supplement habits. None of the subjects are currently taking dietary supplements containing creatine or have taken such diets in a recent study supplementation procedure. The subject's physical characteristics are summarized in Table 6.

  Two days before treatment, the subject was placed in a sitting position, and a preliminary measurement of the maximum voluntary (isometric) contractile force (MVC) of the knee extensor was performed. The MVC was measured using the Macflex system while motivating the subject with immediate image display of force output. For each subject, two trials were performed to measure endurance at 66% MVC, which was sustained until the target force could no longer be maintained despite voice encouragement. After this initial contraction, the subject was allowed to rest on an isometric chair for a rest time of 60 seconds. After the rest period, the second contraction was continued until fatigued. After the second 60 second break, a third contraction until fatigue was performed.

  One day before treatment, subjects reported to the isometric test lab between 8 am and 10 am. MVC was measured and endurance at 66% MVC over 3 contractions was measured at a 60 second break interval as described above. Measurements were made using the subject's dominant foot. A biopsy of the lateral part of the lateral vastus muscle was again taken from the dominant foot.

  On the first day of the treatment study, subjects reported to the blood sampling lab after an overnight fast and at least 8 hours after the last meal. After insertion of the catheter and basal blood sample, each subject followed the replenishment and blood sampling protocol described in Example 3. A dose of 10 mg / kg body weight of β-alanine was administered at 0 hours (9 am), 3 hours, and 6 hours.

  On days 2-15, subjects continued to take 3 doses of 10 mg / kg body weight of β-alanine.

  On the morning of the 14th day, a post-treatment isometric exercise test was performed with the dominant foot to measure MVC and endurance at 66% MVC versus 66% MVC measured on the day before treatment. In the afternoon, a muscle biopsy was taken from the lateral vastus, near the site of the biopsy taken the day before treatment.

  On day 15, the procedure according to day 1 was repeated to measure any overall shift in the plasma concentration profile of β-alanine and taurine over 15 days of supplementation. Plasma β- for 9 hours after ingestion of 10 mg / kg body weight of β-alanine at 0, 3, and 6 hours on days 1 and 15 administered at 3 × 10 mg / kg body weight per day The average change in alanine is shown in FIG.

  One additional subject (number 7) took 3 doses of 10 milligrams / kilogram body weight for 7 days, followed by 3 doses of 20 milligrams / kilogram body weight for 7 days, according to the study. No muscle biopsy was taken from this subject.

  There was no obvious change in muscle carnosine content in the muscles of the six subjects from whom the biopsy was taken. Changes in plasma taurine concentration in 6 subjects reflected changes in β-alanine concentration as described in Example 2.

Table 7 lists endurance measurements at MVC and 66% MVC 1 day before and 14 days after treatment with 3 doses of 10 mg / kg body weight of β-alanine. Average endurance time at 66% MVC increased in 5 of 6 subjects. An increase was also seen in subject 7 who received more.

(Example 5)
The effect of 4 weeks of beta-alanine supplementation using two dosing regimes and equimolar L-carnosine (beta-alanyl histidine) administered over 4 weeks on muscle carnosine content was examined.

Fifteen male subjects aged 20-29 years with no apparent signs of clinical disease and normal height and weight were recruited and investigated (Table 8). All subjects participated in one or more sports and all ate mixed meals containing variable amounts of meat. A record of the approximate amount of meat taken by each subject was recorded during the course of the study.

  Five subjects were assigned to one of three treatment groups (1, 2, and 3). During the study, their diet was supplemented with either β-alanine or carnosine as described in Table 9 (Figure 17). Supplementation was given in soft gelatin capsules containing either 400 mg β-alanine or 500 mg carnosine.

  In Group 1, β-alanine was administered in four divided doses (4 times a day) throughout the day at a fixed rate for 4 weeks.

  Group 2 was administered β-alanine as 8 divided doses throughout the day rather than 4 times in an attempt to maintain a more uniform increase in plasma concentration. In addition, the dose was gradually increased by 800 mg per day per week.

  In group 3, carnosine was again administered in 8 doses in approximately equimolar amounts as in group 2. Therefore, this treatment contained approximately the same amount of β-alanine as group 2 when hydrolyzed to its constituent amino acids.

  Subjects were replenished at the times shown in Table 9 (FIG. 17). Using Bergstroem's percutaneous needle biopsy procedure (1962), a single muscle biopsy of the lateral vastus muscle was taken before and at the end of supplementation. In summary, the procedure involves inserting a hollow needle under aseptically anesthetized condition to obtain a sample of about 20-40 mg containing about 100-700 muscle fibers. The skin and subcutaneous tissue are anesthetized with 1% lignocaine (avoid contact with muscles). An incision is made to the skin and deep fascia with a scalpel blade. Insert a needle without a central rod into the muscle. The muscle mass is pushed inside the window on the side of the needle. The sample is cut by pushing the inner sharp cylinder along the needle. The needle is removed and the central rod is used to eject the sample. Then close the wound.

  Muscle samples from subjects were frozen in liquid nitrogen, lyophilized and analyzed for muscle carnosine and taurine content by HPLC. Table 9 (FIG. 17) shows a breakdown of the dosing schedule adopted for each of the three treatment groups. As a result, there was no physique change in any of the subjects supplemented with β-alanine or carnosine.

(Changes in muscle carnosine (Table 10 and FIGS. 12 and 13))
No significant increase in muscle carnosine content was recorded for group 1 and 3 subjects. In group 2, one subject (number 10) with the highest initial carnosine content (initial carnosine content: 33.3 mmol / kg dry muscle) had no change in muscle carnosine content (later content) : 33.7 mmol / kg dry muscle). When this subject was removed from group 2, this group showed as significant an increase as seen in the other groups. The subject 10 is a person who consumes meat at a medium to high level, but has no other outstanding features.

  Supplementing either β-alanine or carnosine at the same dose (groups 2 and 3) appears to have an equivalent effect in increasing muscle carnosine content.

  This pattern of change is very similar to that observed with the addition of creatine, which may suggest that there is a threshold that is reached quickly with further supplementation with no further effect. In the case of the subject 10, although not being bound by this theory, it seems that the threshold has been reached even before the start of supplementation. However, there is an exception to the upper limit concept. Particularly, the subjects 6 (carnosine after supplementation: 45.9 mmol / kg dry muscle) and the subject 15 (carnosine after supplementation: 68.9 mmol / kg dry muscle). Subjects 6 and 15 had no noticeable features in either their eating habits or participation in exercise.

Table 10 summarizes carnosine muscle concentration data for treatment groups 1-3. Treatment group 2 in italic letters does not include subjects 10 who did not show an increase in muscle carnosine concentration. The initial carnosine concentration of subject 10 was the highest value for all subjects, but may have already been at the “upper limit” level prior to supplementation.

(Changes in muscle β-alanine and histidine (Table 11 and FIG. 14)
The muscle β-alanine concentration was below the detection limit (<0.2 mmol / kg dry muscle) before supplementation and at the end of supplementation. In some subjects, the last dose of β-alanine was given within 1-2 hours of the last muscle biopsy.

  There is no change in muscle histidine concentration with either β-alanine or carnosine supplementation, the latter having the potential to release histidine into the systemic circulation. There was no decrease in histidine concentration for increased carnosine synthesis (each mole requires 1 mole of histidine).

(Changes in muscle taurine (Table 12 and FIGS. 15 and 16)
High concentrations of β-alanine may interfere with the uptake of taurine into the tissue, but the above observations indicate that after administration of both β-alanine and carnosine, increased plasma taurine concentration and urinary taurine This study shows a decrease, and this study has confirmed no decrease in muscle taurine in any of the three groups. Some individuals produced significant changes in muscle taurine content, but both increases and decreases were observed. FIG. 15 is a graph illustrating data showing muscle concentration (average + SD) of taurine before and after supplementation in four different treatment groups. FIG. 16 is a graph for explaining data indicating taurine muscle concentration (average + SD) before and after supplementation in different subjects.

(Conclusion)
These studies illustrate that supplementation with β-alanine and carnosine of the present invention may increase muscle carnosine content. Based on the test results, it appears to be equally effective at increasing carnosine in the tissue.

Changes in muscle buffer capacity help maintain an intracellular microenvironment against H ⁺ accumulation during intense exercise. As such, supplementation with β-alanine or a compound that delivers β-alanine upon ingestion can have a positive effect on athletic performance in sports and general daily activities such as causing lactic acid accumulation. In terms of other chemical activities due to carnosine (as an antioxidant and anti-glycation agent), increasing carnosine concentration can have other beneficial effects apart from those resulting from increased muscle buffering capacity. .

Four weeks of supplementation did not cause any apparent reduction of taurine in the muscle.

  A number of embodiments of the invention have been described. In any case, it will be understood that various modifications can be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the appended claims.

Claims (41)

A method for increasing anaerobic work capacity in an organization, comprising the following steps:
(A) providing a β-alanyl histidine dipeptide and a glycine, insulin, insulin mimetic, or insulin action modifier; and
(B) administering to the tissue β-alanine and at least one of glycine, an insulin mimetic, or an insulin action modifier in an amount effective to increase β-alanyl histidine dipeptide synthesis in the tissue; Increasing the anaerobic working capacity in the organization by,
Including said method. A method for adjusting hydronium ion concentration in a tissue, comprising the following steps:
(A) providing a β-alanyl histidine dipeptide and a glycine, insulin, insulin mimetic, or insulin action modifier; and
(B) administering to the tissue β-alanine and at least one of glycine, an insulin mimetic, or an insulin action modifier in an amount effective to increase the concentration of hydronium ions in the tissue;
Including said method.   2. The method of claim 1, wherein the step of administering β-alanine and at least one of glycine, an insulin mimetic, or an insulin action modifier to the tissue comprises oral administration, administration to blood or plasma, or a combination thereof. Method.   The method of claim 1, wherein the β-alanyl histidine dipeptide comprises carnosine, anserine, or valenin.   A composition comprising an amino acid selected from the group consisting of glycine, insulin, an insulin mimetic, or an insulin action modifier and β-alanine, a chemical derivative of β-alanine, and a peptide containing β-alanine, or an active derivative thereof. And a composition comprising:   6. The composition of claim 5, wherein [beta] -alanine comprises [beta] -alanyl histidine dipeptide.   7. The composition of claim 6, wherein the [beta] -alanyl histidine dipeptide comprises carnosine, anserine, or valenin.   6. The composition of claim 5, further comprising at least creatine or a carbohydrate.   Insulin mimetics include D-pinitol (3-O-methyl-tyrosinositol), 4-hydroxyisoleucine, demethyl-asteriquinone B-1 compound, alpha lipoic acid, R-alpha lipoic acid, guanidinopropionic acid, vanadium compound, vanadium 6. The composition of claim 5, comprising a complex or a synthetic phosphoinositol glycan peptide.   6. The composition according to claim 5, wherein the insulin action modifier is sulfonylurea, thiazolidinedione, or biguanide.   The composition according to claim 5, wherein the composition is a pharmaceutical composition.   The composition according to claim 5, wherein the composition is a dietary supplement or a sports drink.   13. The composition of claim 12, wherein the dietary supplement or sports drink is a human supplement.   At least 0.2, 0.3, 0.4, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or A composition comprising 5 grams of a peptide or ester having β-alanine.   At least 0.2, 0.3, 0.4, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 grams of peptide or ester having β-alanine, Injectable composition.   16. A composition according to claim 14 or 15, wherein the peptide comprises a β-alanyl histidine dipeptide.   17. The composition of claim 16, wherein the [beta] -alanyl histidine dipeptide comprises carnosine, anserine, or valenin.   A human composition comprising at least 200, 250, 300, 450, 500, 550, 600, 650, 700, 750, or 800 mg of β-alanine.   19. The composition of claim 18, wherein the composition is formulated for ingestion or is in an injectable form.   20. A composition according to claim 19, wherein the ingestible formulation is a beverage, gel, food or tablet.   19. The composition of claim 18, wherein the peptide comprises [beta] -alanyl histidine dipeptide.   21. The composition of claim 20, wherein the [beta] -alanyl histidine dipeptide comprises carnosine, anserine, or valenin. A method for increasing the anaerobic capacity of a tissue in a subject comprising the following steps:
(A) (i) an amino acid selected from the group consisting of glycine, insulin, insulin mimetic, or insulin action modifier and β-alanine, a chemical derivative of β-alanine, and a peptide containing β-alanine, or a Preparing a composition comprising a mixture with a composition having an active derivative, (ii) a peptide or ester of at least 0.5 grams of β-alanine, or (iii) at least 200 mg of β-alanine in injectable form Steps to perform, and
(B) administering the composition to the subject in an amount effective to enhance the anaerobic working capacity of the tissue;
And said method comprising:   24. The method of claim 23, wherein the total 24-hour dose of β-alanine is at least 0.2 grams.   24. The method of claim 23, wherein the total 24-hour dose of β-alanine is from about 0.2 grams to about 6.4 grams.   24. The method of claim 23, wherein the composition is given over a period of at least 3 days.   24. The method of claim 23, wherein the composition is provided over a period of at least about 3 days to about 2 weeks.   24. The method of claim 23, wherein [beta] -alanine comprises [beta] -alanyl histidine dipeptide.   29. The method of claim 28, wherein the [beta] -alanyl histidine dipeptide comprises carnosine, anserine, or valenin.   30. The method of claim 28, wherein the total dose over a 24 hour period of beta-alanyl histidine dipeptide is at least about 0.5 grams.   32. The method of claim 30, wherein the total dose of β-alanyl histidine dipeptide over 24 hours is greater than about 5 grams.   30. The method of claim 28, wherein the total dose over 24 hours of [beta] -alanyl histidine dipeptide is from about 5 grams to about 16 grams.   30. The method of claim 28, wherein the total dose over 24 hours of [beta] -alanyl histidine dipeptide is at least 16 grams.   24. The method of claim 23, wherein the composition is administered in multiple doses.   35. The method of claim 34, wherein the composition is administered at least 2 to 8 times over a 24 hour period.   24. The method of claim 23, wherein about 200 mg [beta] -alanine or about 500 mg carnosine is administered about 2 to 8 times per day over a period of several weeks.   24. The method of claim 23, wherein at least about 2 g of β-alanine, or at least about 5 g of carnosine is administered from about 2 to 8 times per day over a period of about 2, 3, or 4 days.   24. The method of claim 23, wherein the amount of composition administered increases daily.   24. The method of claim 23, wherein the amount of composition administered increases weekly.   24. The method of claim 23, wherein the composition is administered for a treatment period lasting at least about 4 weeks. A method for adjusting hydronium ion concentration in tissue in a subject comprising the steps of:
(A) (i) an amino acid selected from the group consisting of glycine, insulin, insulin mimetic, or insulin action modifier and β-alanine, a chemical derivative of β-alanine, and a peptide containing β-alanine, or a Preparing a composition comprising a mixture with a composition having an active derivative, (ii) a peptide or ester of at least 0.5 grams of β-alanine, or (iii) at least 200 mg of β-alanine in injectable form Steps to perform, and
(B) administering the composition to the subject in an amount effective to regulate hydronium ion concentration in the tissue;
Including said method.

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