JP2010031035A6 - Methods and compositions for enhancing anaerobic exercise capacity in tissue - Google Patents

Abstract

Methods and compositions for enhancing anaerobic exercise capacity of muscle and other tissues are provided.
An amount of β-alanine effective to increase β-alanyl histidine dipeptide synthesis in a tissue is supplied to the blood or plasma, and the tissue is exposed to the blood or plasma, thereby β-alanyl histidine A method for controlling the concentration of hydronium ions in the tissue, and a composition for enhancing the anaerobic exercise capacity of the tissue.
[Selection figure] None

Description

  The present invention relates to methods and compositions for enhancing the anaerobic performance of muscles and other tissues.

Background of the Invention Natural food supplements are typically designed to compensate for reduced levels of dietary nutrients in modern humans and animals. In particular, useful auxiliary substances enhance tissue function when ingested. In particular, it is important to supplement certain types of animal diets (the normal diet may be deficient in nutrients available only from meat and animal foods) (eg, human vegetarians and others Animals eat herbivorous foods).

  For example, in sports and competition societies, the importance of natural food supplements that specifically improve athletic performance, such as supplements that promote and strengthen physical robustness for leisure and labor purposes, is increasing. . As another example, anaerobic (eg, lactic acid production) stress can cause the occurrence of fatigue and discomfort that can be experienced with aging. Anaerobic stress can also be caused by increased intramuscular pressure (e.g., during rock climbing, free diving, synchronized swimming, etc.) when the local circulation is partially or completely occluded, etc. It can also occur from a shaku movement. Excess lactic acid production can cause acidification of the intracellular environment.

  Creatine (i.e. N- (aminoiminomethyl) -N-glycine, N-amidinosarcosine, N-methyl-N-guanylglycine, or methylglycosylamine) is characterized by the ability of skeletal muscle and high variable energy requirements Are found in large quantities in other “excitable” tissues (eg, smooth muscle, cardiac muscle, or sperm). Creatine is converted to phosphoryl creatine in an intracellular energy generating biochemical pathway. In mammalian skeletal muscle, the total content of typical creatine (ie, creatine and phosphoryl creatine) can vary from less than 25 mmol per kilogram of fresh muscle to 50 mmol (ie, 3.2-6.5 grams per kilogram of muscle).

  Creatine is produced in the liver and taken up into tissues such as muscle by an active transport system. Creatine synthesis in the body also depends on the intake of creatine present in the meat (eg, 5-10 milligrams per kilogram of body weight per day for an average carnivorous person and almost zero in a vegetarian diet) Will be increased.

  Accumulation of hydronium ions and lactate (anaerobic metabolism) formed during glycolysis significantly increased intracellular pH during intensive exercise or continued exercise under local hypoxia. May decrease. Reduced pH can reduce 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.

  β-alanine and histidine dipeptides and their methylated analogs include carnosine (β-alanyl-L-histidine), anserine (β-alanyl-L-1-methylhistidine), or valenine (β-alanyl-L- 3-methyl histidine). These dipeptides are present in human and other vertebrate muscles. Carnosine is found in significant amounts in, for example, human and equine muscles. Anserine and carnosine are found, for example, in the muscles of dogs, camels, and many avian species. Anserine is the major β-alanyl histidine dipeptide found in many fish. Valenin is the major β-alanyl histidine dipeptide found in several species of aquatic mammals and reptiles. In humans, horses and camels, the highest concentration of β-alanyl histidine dipeptide is found in rapidly contracting glycolytic muscle fibers (types IIA and IIB) used intensively during intense exercise. Lower concentrations are found in oxidative slow contractile muscle fibers (type I). For example, Dunnett, M .; & Harris, R.C. Equine Vet. J. Suppl. 18, see 214-217 (1995). Carnosine is estimated to contribute to the hydronium ion buffering capacity in various muscle fiber types, up to 50% of the total in horse II type fibers.

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

SUMMARY OF THE INVENTION Generally, the present invention relates to methods and compositions for enhancing anaerobic exercise capacity of muscles and other tissues. The method involves the simultaneous accumulation of creatine and β-alanyl histidine dipeptides, or β-alanine and L-histidine analogs in body tissues. The method includes ingesting or injecting the composition into the body. The composition is a mixture of compounds that can increase the availability and absorption of creatine and precursors for the synthesis and accumulation of β-alanyl histidine dipeptide in human or animal muscles. The composition elicits the synthesis and accumulation of β-alanyl histidine dipeptide in the human or animal body when it is introduced into the body.

  The composition comprises a mixture of creatine and β-alanine, creatine, β-alanine and L-histidine, or an active derivative of creatine and β-alanine or L-histidine. Each of β-alanine or L-histidine may be an individual amino acid or may be a component of a dipeptide, oligopeptide or polypeptide. β-alanine and L-histidine may be active derivatives. An active derivative is a compound or precursor thereof derived from a substance that functions in the same or similar manner as the substance in the body, or a compound that is processed into such a substance and placed in the body. Examples include esters and amides.

  In one aspect, the present invention relates to a method for controlling the concentration of hydronium ions in tissue. The method provides a blood or plasma with an amount of β-alanine effective to increase β-alanyl histidine dipeptide synthesis in the tissue and exposes the tissue to the blood or plasma, thereby providing β-alanyl histidine. Increasing the concentration of in the tissue. The method can include supplying an amount of L-histidine to blood or plasma effective to increase β-alanyl histidine dipeptide synthesis.

  In another aspect, the invention relates to a method for increasing anaerobic exercise capacity of a tissue. The method is effective in increasing the synthesis of β-alanyl histidine dipeptide in tissue by supplying the blood or plasma with an amount of β-alanine effective to increase β-alanyl histidine dipeptide synthesis in the tissue. Providing a quantity of L-histidine to the blood or plasma and exposing the tissue to the blood or plasma. The concentration of β-alanyl histidine is increased in the tissue.

  In certain embodiments, the method comprises increasing the concentration of creatine in the tissue. This increasing step may include providing an amount of creatine effective to increase the creatine concentration in the tissue to the blood or plasma (eg, by supplying said amount of creatine to the blood or plasma).

  The providing step of the method can include ingestion or infusion (eg, injection) of the amount of β-alanine or a composition comprising the amounts of β-alanine and L-histidine, or a combination of ingestion and infusion.

  The method can include increasing insulin concentration in blood or plasma. The concentration of insulin can be increased, for example, by injection of insulin.

  The tissue can be skeletal muscle.

  In another aspect, the invention relates to a composition consisting essentially of a peptide source comprising β-alanine, from about 39 to about 99% by weight carbohydrate, and up to about 60% by weight water. The composition comprises about 1 to about 20% by weight β-alanine. The peptide source can include L-histidine. The composition may comprise about 1 to about 20% by weight L-histidine.

  The carbohydrate may be a simple carbohydrate (eg glucose). In another aspect, the invention relates to a composition consisting essentially of a peptide source comprising β-alanine, from about 1 to about 98% by weight creatine source, and up to about 97% by weight water. The composition comprises about 1 to about 98% by weight β-alanine. The peptide source can comprise L-histidine and the composition comprises from about 1 to about 98% by weight L-histidine.

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

  The composition can be a food supplement. The creatine source may be creatine monohydrate.

  The concentration of a component in blood or plasma can be increased by infusion (eg, injection) or ingestion of a substance that can function to cause an increase in plasma concentration. The composition can be ingested in an amount between about 10 grams to about 800 grams per day. This amount can be administered once a day or divided into multiple doses.

  Increased intramuscular content of creatine and β-alanyl histidine dipeptide increases the cell's tolerance to increased hydronium ion production associated with anaerobic exercise and reduces the duration of exercise until fatigue occurs. Can be increased.

  The compositions and methods may be useful to compensate for the loss of these components due to degradation or leaching of β-alanine, L-histidine or creatine during cooking or processing. The compositions and methods may also be useful in replenishing these component deficiencies in a vegetarian diet.

  The method and composition may include β-alanyl histidine dipeptide in, for example, sportsmen, athletes, bodybuilders, synchronized swimming swimmers, soldiers, the elderly, race horses, work and race dogs, competition birds, etc. Can be used to increase, prevent and delay the occurrence of muscle fatigue.

  Other advantages and features of the invention will be apparent from the detailed description and from the claims.

Brief Description of Drawings
After feeding β-alanine and L-histidine for 30 days (100 milligrams per kilogram body weight and 12.5 milligrams per kilogram body weight, 3 times a day, respectively) before feeding and every 2 hours after feeding It is a graph which shows the change of the density | concentration of (beta) -alanine in the plasma. After feeding β-alanine and L-histidine for 30 days (100 mg / kg body weight and 12.5 mg / kg body weight, 3 times a day, respectively) before feeding and every 2 hours after feeding It is a graph which shows the change of the density | concentration of L-histidine in the plasma of each. Horse 1 of 6 horses on the first and last days of 30-day dietary supplementation of β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, respectively) It is a graph which compares and shows the change of the density | concentration of the beta-alanine in the plasma for every time before feeding of -3 and every time after feeding. Horses 4 out of 6 horses on the first and last days of 30-day dietary supplementation of β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, respectively) It is a graph which compares and shows the change of the density | concentration of (beta) -alanine in the plasma for every time before feeding of -6 and every time after feeding. Horse 1 of 6 horses on the first and last days of 30-day dietary supplementation of β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, respectively) It is a graph showing by comparison the change of the density | concentration of L-histidine in the plasma for every time before feeding of -3 and every time after feeding. Horses 4 out of 6 horses on the first and last days of 30-day dietary supplementation of β-alanine and L-histidine (100 mg / kg body weight and 12.5 mg / kg body weight, respectively) It is a graph which represents the change of the density | concentration of L-histidine in the plasma for every time before feeding of -6, and every time after feeding. Horses by beta-alanine and L-histidine for 30 days (100 milligrams per kilogram of body weight and 12.5 milligrams per kilogram of body weight, 3 times a day, respectively) on the first and last days, before and after feeding It is a graph which compares and shows the change of the average density | concentration (n = 6) of (beta) -alanine in plasma. Horses by beta-alanine and L-histidine for 30 days (100 milligrams per kilogram of body weight and 12.5 milligrams per kilogram of body weight, 3 times a day, respectively) on the first and last days, before and after feeding It is a graph which compares and shows the change of the average density | concentration (n = 6) of L-histidine in plasma. Increased carnosine concentration in type II skeletal muscle fibers of six purebred horses (average of total IIA and IIB fibers) and plasma β-alanine concentration-time curve between days 1 and 30 of supplementation an increase in the area _{(AUC (0-12hr))} over the first 12 hours of the day under a graph showing the correlation. It is a graph which shows the average result of the administration to the test subject of (beta) -alanine, a broth, or carnosine. FIG. 6 is a graph showing the mean change in plasma β-alanine over 9 hours of treatment. 2 is a graph showing the mean change in plasma β-alanine over 9 hours after oral intake of 10 milligrams β-alanine per kilogram body weight. 2 is a graph showing the mean (n = 6) plasma β-alanine concentration over 24 hours on day 1 and day 30 of the treatment period.

DESCRIPTION OF THE PREFERRED EMBODIMENTS β-Alanylhistidine dipeptides such as carnosine, anserine, and valenin have pKa values between about 6.8 and 7.1 and are involved in controlling intracellular pH homeostasis during muscle contraction and fatigue development. ing. The content of other substances involved in buffering hydronium ions, such as amino acid residues in proteins, inorganic and organic phosphates and bicarbonates, is constrained by their involvement in other cellular functions . β-alanyl histidine dipeptides provide an effective way to accumulate pH sensitive histidine residues in cells. Changes in muscle β-alanyl histidine dipeptide concentration affect an individual's anaerobic performance.

  β-alanyl histidine dipeptide is synthesized in the body from β-alanine and L-histidine. These precursors are made available in the diet, including those from the degradation of ingested β-alanyl histidine dipeptides produced in the body.

  Β-alanine in the body is transported to tissues such as muscles. Under typical feeding conditions, the concentration of β-alanine is low compared to the concentration of L-histidine in human and horse plasma. These concentrations must be considered in relation to the affinity of the carnosine synthetase carnosine synthetase for its substrate 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 1000-2300 μM. The low affinity of carnosine synthetase for β-alanine and the low concentration of β-alanine in muscle indicate that the synthesis of β-alanyl histidine dipeptide is limited by the concentration of β-alanine in muscle.

  Increasing the amount of β-alanyl histidine dipeptide in muscle favorably affects muscle performance and the amount of exercise achieved by the muscle. Accordingly, the synthesis and accumulation of β-alanyl histidine dipeptide is increased in human or animal tissues.

  The synthesis and accumulation of β-alanyl histidine dipeptides in humans or animals can include increased blood or plasma concentrations of β-alanine, increased blood or plasma concentrations of β-alanine and creatine, or β-alanine, L-histidine and By increasing the blood or plasma concentration of creatine, it can be increased with increasing creatine content in the body. The increase in dipeptide can occur simultaneously with the increase in β-alanine concentration.

  Plasma concentrations of β-alanine, L-histidine and creatine can be increased by ingestion or infusion of β-alanine, L-histidine and creatine or their active derivatives. The composition can be administered orally, enterally, or parenterally. β-alanine and creatine or β-alanine, L-histidine and creatine are preferably taken orally.

  The composition may include carbohydrates (eg, simple carbohydrates), insulin, or substances that stimulate the production of insulin.

  The composition can be ingested as a food supplement. Preferably the composition is administered one or more times per day. The dose of β-alanine can be between about 5 milligrams and about 200 milligrams per kilogram of body weight. The dose of creatine (eg, creatine monohydrate) can be between about 5 milligrams and 200 milligrams per kilogram body weight. The dose of L-histidine can be between about 1 milligram and 100 milligrams per kilogram of body weight.

  The dosage of a simple carbohydrate (eg glucose) can be between about 0.5 and 2.0 grams per kilogram of body weight.

  For an 80 kilogram human, a suitable daily dose is 0.4 grams to 16.0 grams β-alanine, 0.4 grams to 16.0 grams creatine monohydrate, 0.08 grams to 8.0 grams L-histidine, or It can be 40 grams to 160 grams of glucose or other simple carbohydrate. The composition can be a solid or liquid dosage form for ingestion, or a suspension dosage form, or a liquid or suspension dosage form for injection into the body. The composition is ingested by humans in an amount between 2 grams and 1000 grams per day (for example, an amount between 10 grams and 800 grams), which can be taken in portions during the day. it can. Daily intake in animals will be adjusted for body weight.

  For humans and animals, the composition comprises (a) 1% to 99% by weight β-alanine, 1% to 99% by weight creatine monohydrate, and 0% to 98% by weight water; (b) 1 wt% to 98 wt% β-alanine, 1 wt% to 98 wt% L-histidine, 1 wt% to 98 wt% creatine monohydrate, and 0 wt% to 97 wt% Water; (c) 1% to 20% β-alanine, 39% to 99% glucose or other simple carbohydrate, and 0% to 60% water, or (d) 1% % To 20% β-alanine, 1% to 20% L-histidine, 39% to 99% glucose or other simple carbohydrate, and 0% to 60% water, can do.

  The following are specific examples of methods and composition methods for enhancing the anaerobic performance of muscles and other tissues.

Example 1
The effect of multiple daily supplementation of β-alanine and L-histidine on a standard diet on carnosine concentrations in type I, IIA and IIB skeletal muscle fibers of purebred horses was evaluated. Feed normal experimental 4-9 year old purebred horses (3 young female horses and 3 castrated horses) for 1 month (30 days) (pre-supply period) prior to the start of the supply period Conditioned. During the feeding conditioning period, each horse receives 1 kilogram of shaped feed (Spillers Competitive Horse Cube) and 1 kilogram of ground sugar beet pulp as a source of complex and simple carbohydrate three times a day (each 08: 30, 12:30 and 16:30). Crushed hay (3 kg dry weight) was also given twice daily (at 09:00 and 17:00). I was allowed to drink water freely.

  The same feeding regime was implemented during the supplementation period. However, β-alanine and L-histidine (free base) were supplemented when giving a hard feed. β-alanine and L-histidine were mixed directly into the standard diet. Individual doses of β-alanine and L-histidine were calculated according to body weight. β-alanine was administered at 100 mg / kg body weight, and L-histidine was administered at 12.5 mg / kg body weight. Feeding began on day 1 of this protocol and ended on day 30. Blood samples (5 milliliters) were taken on days 1, 6, 18, 24 and 30 and heparin was added. On days 1 and 30, blood samples were taken every hour for the total 12 hours before the first feeding. On the remaining 3 days of blood collection, blood was collected before the first feeding and 2 hours after each subsequent feeding. On the day before the start of replenishment (Day 0), after local anesthesia of the skin, a muscle biopsy was taken from the right intermediate muscle (m. Glutaeusmedius) of each horse using a Bergstrom-Stille percutaneous biopsy needle. Subsequent muscle biopsies were taken as close as possible to the original collection site and immediately after the replenishment period (day 31). Horses were clinically monitored daily. This monitoring included gross inspection and body weight measurements, twice daily rectal temperature measurements, and weekly blood collections for clinical biochemistry and hematology. During the experiment, the horses did not receive formal training or exercise, but were allowed to exercise freely for 1 hour every day.

  Individual muscle fiber fragments dissected from lyophilized muscle biopsy material are described in Kaiser and Brcok, Arch. Neurol., 23: 369-379 (1970), by a histochemical staining of myosin ATPase activity at pH 9.6, followed by preincubation at pH 4.50, to give I, IIA Or characterized as either type IIB.

  Heparinized plasma samples were extracted and analyzed for β-alanine and L-histidine concentrations by high performance liquid chromatography (HPLC). Weighed individual muscle fibers were extracted and Dunnett and Harris, “High Performance Liquid Chromatographic Measurement of Imidazole Dipeptides, Histidine, 1-Methylhistidine, 3-Methylhistidine in Muscle and Individual Muscle Fibers”, J. Am. Chromatogr. B. Biomed. Carnosine was analyzed by HPLC according to the method described in Appl., 688: 47-55 (1997).

  The difference in carnosine concentration contained in the fiber type before and after replenishment was determined in horses using one-way analysis of variance (ANOVA). When the difference was recognized, the significant difference was calculated | required by the multiple comparison test (Fisher's PLSD).

  Addition of β-alanine and L-histidine to the diet did not cause any taste problems. During the 30-day supplementation period, no adverse physiological or behavioral effects of supplemental diet were observed in any horse. No significant change in body weight was recorded and rectal temperature remained within the normal range. There were no acute or chronic changes in clinical biochemistry or hematology. Β-alanine was not detected in the plasma of horses before supplementation. The lower limit of β-alanine quantification in plasma by the assay used was 3 micromolar (μM). The plasma L-histidine concentration of 6 horses before the start of supplementation was 36.6-54.4 μM.

  Individual changes in plasma β-alanine and L-histidine concentrations over all blood collection days are shown in FIGS. 1 and 2 for 5 of 6 horses, respectively. As the supplementation period increased, pre-feed concentrations of plasma β-alanine and L-histidine tended to increase. Furthermore, the plasma concentration response to supplementation also increased over the 30 day supplementation period. This response was greater with β-alanine.

A comparison of the changes in plasma β-alanine and L-histidine concentrations for the day before the first supplementation and for the first and last days of the supplementation period is shown in Figures 3a and 3b and 6a for each of the six horses. Shown in 4b. FIG. 5 shows the mean (± SD) change (n = 6) in plasma β-alanine concentration over time over 24 hours on the first day (day 1) and the last day (day 30) of the supplementation period. The area under the mean plasma β-alanine concentration (AUC _{(0-24 hr)} ) for the time curve over 24 hours was even greater on the 30th day of supplementation.

The mean (± SD) change (n = 6) in plasma L-histidine concentration over time over 24 hours from the first day (day 1) and the last day (day 30) of the supplement period is shown in FIG. The area under the mean plasma β-alanine concentration (AUC _{(0-24 hr)} ) for the time curve over 24 hours was greater on the 30th day of supplementation. The larger AUC of plasma β-alanine on the last day of supplementation (day 30) compared to the first day of supplementation (day 1) increases the absorption of β-alanine from the horse's gastrointestinal tract as supplementation proceeds Suggests to do. Similar effects were observed with respect to changes in plasma L-histidine concentration during the supplementation period. Maximum plasma concentrations of β-alanine and L-histidine appeared in each case about 1-2 hours after feeding.

* 補給前と有意に異なる、p<0.05
** 補給前と有意に異なる、p<0.01
*** 補給前と有意に異なる、p<0.005 A total of 397 skeletal muscle fibers (192 before supplementation; 205 after supplementation) were excised from six horses and analyzed for carnosine. Mean (± SD) carnosine concentration (mmol kg ⁻¹ dw) per kilogram of dry weight in skeletal muscle fibers of type I, IIA and IIB before and after supplementation obtained from each of the six horses Are shown in Table 1, where n is the number of individual muscle fibers analyzed. 30 days after supplementation with β-alanine and L-histidine, the average carnosine concentration in type IIA and IIB fibers increased in all six horses. These increases were statistically significant in 7 cases. The increase in mean carnosine concentration in type IIB skeletal 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.

* Significantly different from before replenishment, p <0.05
** Significantly different from before replenishment, p <0.01
*** Significantly different from before replenishment, p <0.005

Table 2 shows the absolute increase (eg, mmol kg ⁻¹ dw) and percent increase in mean carnosine concentration in type IIA and IIB skeletal muscle fibers obtained from 6 horses.

It was observed that horses that showed a greater increase in muscle carnosine concentration after 30 days of supplementation also showed a greater increase in plasma β-alanine AUC between days 1 and 30 of the supplementation period. Referring to FIG. 7, mean carnosine concentration averaged between type IIA and type IIB skeletal muscle fibers and plasma β-alanine AUC (AUC _(0-12hrs) over the first 12 hours on days 1 and 30 of supplementation. ₎ ) And a significant correlation (r = 0.986, p = 0.005) was observed for 5 out of 6 horses. Only 5 horses were used to determine the regression line. Horse 6 (filled circles) showed no appreciable increase until the last day of supplementation, higher than the increase in plasma β-alanine concentration observed on day 1. This horse was different from the other 5 horses that showed an incremental increase on each blood collection day. For these reasons, horse 6 was excluded from the regression equation.

An increase in muscle carnosine concentration after 30 days of supplementation with β-alanine and L-histidine will cause a direct increase in overall muscle buffer capacity. This increase can be calculated using the Henderson-Hasselbach equation. Table 3 shows the calculated values for the increase in muscle buffering capacity of IIA and IIB type skeletal muscle fibers in 6 purebred horses.

Example 2
The effect of multiple daily supplements of β-alanine and L-histidine on the diet was evaluated on the carnosine content of human I, IIA and IIB skeletal muscle fibers. The plasma concentration of β-alanine in 6 normal subjects was monitored after ingesting broth supplying approximately 40 mg / kg (body weight) of β-alanine. A dose of 10 and 20 mg / kg (body weight) of β-alanine was also administered.

  Broth was prepared as follows. Fresh chicken breast (skinned and boned) was minced and boiled for 15 minutes with water (1 liter per 1.5 kg of chicken). The remaining chicken was removed by coarse filtration. The filtrate was seasoned with carrot, onion, celery, salt, pepper, basil, parsley and tomato puree, reboiled for another 15 minutes, cooled and then passed through fine muslin at 4 ° C. for final filtration. 870 mL broth was obtained from 1.5 kg chicken and 1 L water. A portion of this stock was assayed for its total β-alanyl-dipeptide content (eg, carnosine and anserine) and β-alanine. A typical analysis was as follows.

Total β-alanyl-dipeptide 74.5 mM
Free β-alanine 5.7 mM
The six male subjects were in normal health and the age ranged from 25 to 53 years as shown in Table 4. The experiment was started after an overnight fast (eg, at least 12 hours after the last meal containing meat). Subjects were given the freedom to drink a small amount of hot water before the start of the experiment. The catheterization started at 08:30 and the experiment started at 09:00.

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

  In one session, 8 mL / kg (body weight) broth containing about 40 mg / kg (body weight) of β-alanine (eg, in the form of anserine and carnosine) was ingested. In a subject weighing 75 kg, this corresponds to ingesting 600 mL of broth containing 3 g of β-alanine. In another session, 3 mL / kg (body weight) of fluid containing a test amount of β-alanine was ingested with an additional 5 mL / kg (body weight) of water. In all sessions, subjects drank another 8 mL / kg (body weight) of water (50 mL each) within 1-2 hours after ingestion. Six hours later, I was given a vegetarian pizza. After 8 hours, she returned to regular diet.

  2.5 mL venous blood samples were taken from the indwelling catheter every 10 minutes for the first 90 minutes and then 120, 180, 240 and 360 minutes. Blood samples were distributed into tubes containing lithium-heparin as an anticoagulant. The catheter was maintained by flushing with saline. Jones & Gilligan (1983) Chromatogr. Plasma samples were analyzed by HPLC according to the method described in 266: 471-482 (1983).

Table 4 summarizes the treatment assignments during the β-alanine absorption experiment. The estimated equivalent amount of β-alanine is shown in Table 3.

  The plasma concentration curve after each treatment is shown graphically in FIG. Average results of administration of β-alanine, broth or carnosine according to the treatment regimen in Table 4. Neither carnosine nor anserine was detected in plasma after ingestion of chicken broth or after other treatments.

  Ingestion of broth resulted in a maximum plasma concentration of 427.9 (SD 161.8) μM. When one subject received carnosine equal to 20 mg / kg (body weight) of β-alanine, plasma β-alanine concentrations increased as well.

  Administration of all treatments except the control increased plasma taurine levels. Changes in taurine concentration closely reflected changes in β-alanine concentration. Administration of broth, a natural food, caused a similar increase in plasma taurine, indicating that such a response occurs normally after most meals.

Example 3
What is the effect on the plasma concentration profile of β-alanine and taurine when 10 mg / kg (body weight) of β-alanine is administered three times a day (ie morning, noon, and night) for 7 days? I investigated. Three subjects were administered 10 mg / kg (body weight) of β-alanine three times a day for 7 days, and the plasma concentration profiles at the beginning and end of the 7 days were examined.

The subjects were three normal healthy men aged 33-53. Subjects were administered 10 mg / kg (body weight) of β-alanine three times a day for 8 days. Two of the subjects were subsequently administered 20 mg / kg (body weight) of β-alanine three times a day for a further 7 days (from day 9 to day 15). Subjects were sent to the blood collection room on the 1st day (before any treatment), 8th day, and 15th day at 8am after fasting overnight. Subjects were asked not to take any meat-containing meals for 12 hours prior to this study. On each of these three test days, subjects were placed with a catheter and the first blood sample was taken when β-alanine was administered at or near 9 am, noon and 3 pm. Blood samples were taken after 30, 60, 120 and 180 minutes and analyzed for changes in plasma concentrations of β-alanine and taurine. On each test day, a 24-hour urine sample was collected and analyzed by HPLC to measure the excretion amounts of β-alanine and taurine. A summary of the treatment is shown in Table 5.

  Plasma β-alanine concentrations are summarized in FIG. With each administration, the maximum β-alanine concentration was reached at 1/2 hour or 1 hour after administration and then decreased to a basal level of 0-10 μM at 3 hours (ie immediately before the next administration). As shown by the area under the plasma concentration curve (AUC), the response on treatment day 8 tended to be lower than on day 1.

Example 4
Isometric endurance at 66% of muscle carnosine content and maximum voluntary contractility when administered 40 mg / kg (body weight) β-alanine three times a day (ie morning, noon, and night) for 2 weeks We investigated what effect it had on force.

Subjects were recruited 6 normal men aged 25-32 years with no evidence of metabolic or muscle disease. Subjects were asked about their recent diet and supplementary habits. None of the subjects recently took supplements containing creatine, and no one did so in a recent pilot supplement. The subject's physical characteristics are summarized in Table 6.

  Two days prior to treatment, the subject's maximum voluntary (isometric) contractile force (MVC) in a sitting state was preliminarily measured. MVC was measured using the Macflex system while guiding the subject by immediately displaying the MVC output. In each subject, two trials were performed and the endurance at 66% MVC was measured, with 66% MVC being maintained until it was no longer possible to maintain the target force regardless of cheering. After this initial contraction, the subject was allowed to sit in an isometric chair for a 60 second break. After the rest period, the second contraction was continued until fatigued. After giving a second break for 60 seconds, the third contraction was continued until fatigued.

  One day prior to treatment, subjects were sent to an isometric laboratory between 8-10 am. In addition to measuring MVC, three contractions were performed with a 60 second rest interval as described above, and endurance at 66% MVC was measured. The measurement was performed using the subject's dominant foot. A biopsy of the outer part of the lateral vastus muscle was again taken from the dominant foot.

  After at least 12 hours of fasting overnight and the last meat-containing meal, subjects were sent to the blood collection room at 8 am on the first day of treatment. Each subject was placed a catheter and the first blood sample was collected, followed by the replenishment and blood collection protocol described in Example 3. 10 mg / kg (body weight) of β-alanine was administered at 0 hours (9 am), 3 hours and 6 hours.

  On days 2-15, the subject continued to receive 10 mg / kg (body weight) of β-alanine three times.

  On the morning of day 14, an isometric exercise test after treatment was performed on the dominant foot to measure MVC and to measure endurance at 66% MVC relative to 66% MVC measured before treatment. That afternoon, biopsies of the outer vastus muscle were collected from the vicinity of the site from which the biopsy was taken prior to treatment.

  On day 15, the procedure performed on day 1 was repeated to determine the overall change in the plasma concentration profile of β-alanine and taurine over 15 days of supplementation. Plasma for 9 hours after oral administration of 10 mg / kg (body weight) of β-alanine at 0, 3 and 6 hours on days 1 and 15 when a dose of 10 mg / kg (body weight) was administered 3 times a day The average change of β-alanine is shown in FIG.

  One additional subject (No. 7) was administered a dose of 10 mg / kg (body weight) 3 times for 7 days, followed by a dose of 20 mg / kg (body weight) 3 times for 7 days did. No muscle biopsy material was collected from this subject.

  There was no apparent change in muscle carnosine content in muscle biopsies taken from 6 subjects. As described in Example 2, the change in plasma taurine concentration in 6 subjects reflected the plasma β-alanine concentration.

Table 7 shows the measured endurance in MVC and 66% MVC one day before and 14 days after the administration of 10 mg / kg (body weight) of β-alanine three times. Average endurance at 66% MVC increased in 5 of 6 subjects. In addition, an increase was also observed in subject 7 who was administered a higher dose.

  Other embodiments are within the scope of the claims.

Claims (25)

  An amount of β-alanine effective to increase β-alanyl histidine dipeptide synthesis in the tissue is supplied to the blood or plasma, and the tissue is exposed to the blood or plasma, thereby reducing the concentration of β-alanyl histidine. A method of controlling the concentration of hydronium ions in a tissue comprising increasing in the tissue.   2. The method of claim 1, further comprising providing the blood or plasma with an amount of L-histidine effective to increase β-alanyl histidine dipeptide synthesis in the tissue.   2. The method of claim 1, further comprising increasing the concentration of creatine in the tissue. 4. The method of claim 3, further comprising providing the blood or plasma with an amount of L-histidine effective to increase β-alanyl histidine dipeptide synthesis in the tissue.   4. The method of claim 3, wherein increasing the concentration of creatine in the tissue comprises providing the blood or plasma with an amount of creatine effective to increase the concentration of creatine in the tissue.   6. The method of claim 5, further comprising providing the blood or plasma with an amount of L-histidine effective to increase β-alanyl histidine dipeptide synthesis in the tissue.   The method of claim 1, wherein the feeding step comprises ingesting a composition containing the amount of β-alanine.   The method of claim 1, wherein the feeding step comprises injecting a composition containing the amount of β-alanine.   2. The method of claim 1, further comprising increasing the concentration of insulin in blood or plasma.   10. The method of claim 9, further comprising providing the blood or plasma with an amount of L-histidine effective to increase β-alanyl histidine dipeptide synthesis in the tissue.   The method of claim 1, wherein the tissue is skeletal muscle.   The method of claim 1, wherein the tissue is human tissue.   The method of claim 1, wherein the tissue is animal tissue.   Supplying blood or plasma with an amount of β-alanine effective to increase β-alanyl histidine dipeptide synthesis in the tissue and an amount of L-histidine effective to increase β-alanyl histidine dipeptide synthesis A method for enhancing an anaerobic exercise capacity of a tissue comprising supplying the blood or plasma and exposing the tissue to the blood or plasma, thereby increasing the concentration of β-alanylhistidine in the tissue.   15. The method of claim 14, further comprising increasing the concentration of creatine in the tissue.   15. The method of claim 14, wherein the feeding step comprises ingesting a composition containing the amount of β-alanine and the amount of L-histidine.   15. The method of claim 14, wherein the feeding step comprises injecting a composition containing the amount of β-alanine and the amount of L-histidine.   15. The method of claim 14, further comprising increasing the concentration of insulin in blood or plasma.   15. The method of claim 14, wherein the tissue is skeletal muscle.   15. The method of claim 14, wherein the tissue is human tissue.   15. The method of claim 14, wherein the tissue is animal tissue.   A composition consisting essentially of a peptide source comprising β-alanine, about 39 to about 99% by weight carbohydrate, and up to about 60% by weight water, comprising about 1 to about 20% by weight β-alanine. Said composition comprising.   23. The composition of claim 22, wherein the peptide source comprises L-histidine and the composition comprises from about 1 to about 20% by weight L-histidine.   A composition consisting essentially of a peptide source comprising β-alanine, about 1 to about 98% by weight creatine source, and up to about 97% by weight water, comprising about 1 to about 98% by weight β-alanine A composition as described above, comprising:   25. The composition of claim 24, wherein the peptide source comprises L-histidine and the composition comprises from about 1 to about 98% by weight L-histidine.

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