HK1144549A - Anti-fatigue agent comprising amino acid composition - Google Patents

Description

Anti-fatigue agent containing amino acid composition

Technical Field

The present invention relates to an anti-fatigue agent containing an amino acid composition comprising a specific type and a specific amount of an amino acid.

Background

For human beings living in modern society, the phenomenon of "fatigue" has become a condition that is not negligible. According to the results of recent investigation, nearly 60% of all nations in japan feel fatigue, of which nearly 40% have continued for 6 months or more. When converted into an employment population, approximately 3000 thousands of workers work with a continuous feeling of fatigue. In addition, it causes a problem in daily life when severe chronic fatigue occurs. Further, the economic loss due to such fatigue is estimated to be on the scale of several billions of yen to one trillion of yen, including the cost associated with the recovery measures.

When fatigue is classified according to its performance, it is roughly classified into acute fatigue and chronic fatigue. The former occurs in units of minutes to hours, and most of them can be recovered by a short time of rest. The latter is an acute fatigue accumulation, requiring days for recovery, weeks for the long, and in the case of a long period, more than 6 months.

When the classification is made based on the fatigue-producing site, the classification is roughly classified into muscle fatigue (physical fatigue) and nerve fatigue (mental fatigue). However, although such clear classification into muscle fatigue and nerve fatigue is possible, they are often in fact complexly correlated from the viewpoint of stress (stress). Therefore, the above simple classification cannot clearly find a processing method, which is the present situation. On the other hand, if scientific knowledge of their interrelationships is available, it would be helpful to construct effective fatigue recovery measures.

Various methods have been studied for preventing or recovering from fatigue, including a simple decompression method such as bathing and a therapeutic method by administering a drug. These methods aim to recover from fatigue of each of the muscle fatigue and the nerve fatigue, but actually, there is no method for obtaining a fatigue recovery that can satisfy both of them.

In addition, these methods may include administration of a pharmaceutical agent, nutritional supplement, or the like. When administering a drug, it is troublesome to require a doctor or the like to make a diagnosis or a prescription. Since the method of administering foods such as nutritional supplements is simple and easy to acquire in daily life, research and development of the foods and development of products have been advanced in recent years. The market for nutritional supplements is continuously expanding, and nutritional supplements for fatigue recovery, such as those represented by citric acid, vitamins, and coenzyme Q10, are sold even in convenience stores and the like. However, the effects of fatigue recovery on the above-mentioned muscle fatigue and nerve fatigue are not clearly described in these nutritional supplements, and a method of satisfying both of them at the same time has not been actually obtained.

However, a product composed of 17 amino acid compositions contained in saliva secreted from larvae of vespid wasps (hornets) and known under the trade name "VAAM" has received attention for its effect of mainly improving exercise ability. This "VAAM" also has an effect of recovering fatigue, as mentioned in patent document 1, for example. In addition, development of various amino acid compositions derived from this technique has been advanced, and for example, patent document 2 discloses an amino acid composition comprising 12 amino acid compositions for rapidly recovering mental fatigue such as muscle fatigue and a feeling of lassitude accompanied therewith.

Further, for example, patent document 3 discloses that valine, leucine, and isoleucine known as BCAA have an effect of preventing central nervous system fatigue (brain fatigue) or recovering brain fatigue in addition to an application of improving motor ability, and discloses a central nervous system fatigue preventing/recovering agent obtained from these amino acids.

However, none of these conventional amino acid compositions has been sufficiently studied for the purpose of preventing both muscular fatigue and nerve fatigue, and there are industrial (manufacturing) and economic problems because many kinds of amino acids and a large amount of amino acids need to be blended, and amino acids which are difficult to dissolve and troublesome to prepare, amino acids which are high in cost, and the like are contained.

On the other hand, as a result of some studies on fatigue, it is found that a decrease in blood concentration of a certain amino acid or entry into a specific tissue occurs with fatigue. For example, non-patent document 1 discloses that amino acids such as proline, glycine, and alanine are significantly consumed and reduced in humans subjected to a long-term load from automobiles.

In addition, non-patent document 2 studies the change in the concentration of each component in plasma with or without supplementation with a carbohydrate during exercise. The results showed that the concentrations of glycine, alanine, lysine, threonine and histidine were reduced in plasma, and particularly the reduction rate of the concentration of histidine was large.

In addition, non-patent document 3 also studies changes in plasma concentrations of various components when exercise is performed or not performed. The results showed that the tryptophan concentration was particularly greatly reduced.

Although the causal relationship between the decrease in the concentration of each amino acid and the phenomenon of exercise or fatigue is not clear, it is presumed that these amino acids are substances that are consumed in the body in some way in the cycle of energy metabolism. Therefore, it is considered that muscle fatigue can be recovered by supplementing amino acids at a visually reduced concentration. However, after muscle fatigue and nerve fatigue, fatigue of both cannot be recovered by simply supplementing these amino acids. Further, it has not been known that these amino acids are supplemented before muscle fatigue and nerve fatigue to prevent and further recover from fatigue of both.

Patent document 1: japanese patent No. 2518692

Patent document 2: japanese laid-open patent publication No. 8-198748

Patent document 3: japanese re-listing 2002/034257

Non-patent document 1: G. ア - ボルグ et al (G.Ahlborg et al), The Journal of clinical Investigation, Vol.53, No. 4 1974, p.1080-1090

Non-patent document 2: T.L. バザ - レ et al (T.L. Bazzaree et al), Journal of the American College of Nutrition, 1992, Vol.11, No. 5, p.501-511

Non-patent document 3: A.H. フオ - スランド et al (A.H. Forslund et al), am.J. physiol Endocrinol Meteb, 2000, Vol.278, p.857-867

As described above, the conventional amino acid compositions are mainly intended to improve exercise performance, and studies from the viewpoint of the anti-fatigue or anti-fatigue effect are not sufficient. Further, although there is a technique for particularly claiming an effect of recovering from fatigue or preventing fatigue, studies from the viewpoint of an effect of preventing both muscular fatigue and nerve fatigue are not sufficient, and it is difficult to say that the effects have been solved. In addition, these conventional amino acid compositions require a large number of amino acids to be prepared into a composition, and thus have problems that the preparation thereof is troublesome and the cost thereof inevitably increases.

Disclosure of Invention

As a result of intensive studies, the present inventors have found that, by constituting an amino acid composition in a composition and amount ratio which have not been achieved in the past, a conventionally insufficient effect of preventing fatigue of both muscle fatigue and nerve fatigue can be obtained, and have completed the present invention through further studies.

Accordingly, an object of the present invention is to provide an anti-fatigue agent containing an amino acid composition comprising a specific type of amino acid and a specific amount of amino acid.

That is, the present invention relates to an anti-fatigue agent containing an amino acid composition comprising the following amino acids

30-200 parts of proline;

60-140 parts of glycine;

50-260 parts by weight of alanine;

50-130 parts of lysine;

30-75 parts by weight of tryptophan;

20-40 parts of histidine.

The present invention also relates to the above anti-fatigue agent, wherein the amino acid composition further contains the following amino acids:

3-75 parts of tyrosine;

15-45 parts of arginine.

The present invention also relates to the above anti-fatigue agent, wherein the amino acid composition further contains the following amino acids:

30-55 parts by weight of valine;

35-60 parts by weight of leucine;

25-60 parts of isoleucine.

The present invention also relates to the above anti-fatigue agent, wherein the amino acid composition as an active ingredient is administered in the range of 0.01 to 8 g/kg/day.

Furthermore, the present invention relates to the above anti-fatigue agent, which prevents both muscle fatigue and nerve fatigue at the same time.

The present invention also relates to the above anti-fatigue agent, wherein muscle fatigue is evaluated by an activity measurement, and nerve fatigue is evaluated by a blood biomarker measurement.

The present invention also relates to the above anti-fatigue agent, wherein the effect of preventing muscle fatigue and nerve fatigue is evaluated when the amount of action of the non-administered group is 100 and the amount of action (relative value) of the administered group is 110 or more in the evaluation by the measurement of the amount of action, and when the measured concentration of the non-administered group is 100 and the measured concentration (relative value) of the administered group is 96 or less in the evaluation by the measurement of the blood biomarker.

The composition used in the anti-fatigue agent of the present invention is an amino acid composition having a composition and a content ratio which have not been achieved before. This can provide a high effect in preventing fatigue, which has not been sufficient in the past. That is, according to the present invention, both actual muscle fatigue and nerve fatigue can be prevented at the same time for the first time.

In the present invention, 2 systems, specifically, the measurement of the amount of movement during fatigue and the measurement of a blood biomarker during fatigue, are used as evaluation systems, and these are used as indicators of muscle fatigue and nerve fatigue, respectively. As a result, the amino acid composition of the present invention can obtain a particularly significant fatigue prevention effect in both evaluation systems.

Further, according to the present invention, since a high fatigue preventing effect is obtained by using 6 to 11 kinds of amino acids less than the conventional amino acid composition, the kinds of raw materials required therefor are reduced, the production trouble is reduced, and a high effect is exerted industrially and economically.

Further, according to the present invention, since a high fatigue preventing effect is obtained by using a predetermined amount of amino acid smaller than that of the conventional amino acid composition, the present invention also exerts a high effect industrially and economically. Further, when the preparation is carried out in such a manner that the effect equivalent to that of the conventional preparation is obtained, since the amount of each amino acid used is small, the preparation is carried out in order to adapt the preparation to a beverage or the like, and the reduction of the volume of the beverage or the like can be achieved, and the preparation is effective particularly for a portable beverage such as a sports drink.

The present inventors paid attention to proline, glycine, alanine, lysine, tryptophan, and histidine, which are amino acids whose concentration in blood decreases during exercise, and since these amino acids affect some reactions in the body, they have studied to add these amino acids at a predetermined concentration in order to supplement blood with these amino acids.

Although details of the reaction of the above amino acids in the body are not clear, it is considered that proline, for example, is used as an energy source for muscle and has an effect of burning fat and an effect of inhibiting sympathetic nerve activity; alanine is used as a muscle energy source and has the effect of promoting sympathetic nerve activity; lysine has effects of promoting proper metabolism of fatty acid, improving attention, promoting sympathetic nerve activity, etc.

In addition, since tyrosine, arginine, valine, leucine, and isoleucine, which are amino acids that increase in blood concentration during exercise, affect some in vivo reactions, it has been studied to add these amino acids at a predetermined concentration in order to supplement them to the body outside the blood.

Although details of the reaction of the above-mentioned amino acids in the body are not clear, for example, arginine is considered to have an action of promoting protein synthesis in muscle and promoting sympathetic nerve activity; valine is an essential component contained in protein of muscle, and has the effect of promoting sympathetic nerve activity; leucine has the effects of improving muscle strength and promoting sympathetic nerve activity; isoleucine is present in muscle proteins, and is used as an energy source for muscles, and has an effect of inhibiting sympathetic nerve activity.

Further, it is considered that these tyrosine, arginine, valine, leucine, isoleucine are classified into valine, leucine, isoleucine which is a Branched Chain Amino Acid (BCAA), and tyrosine, arginine which is not BCAA.

Drawings

Fig. 1 is a graph showing a test schedule of the fatigue resistance effect verification test.

Detailed Description

The present invention will be described in detail below, but the present invention is not limited to the embodiments described below.

The amino acid composition contained in the anti-fatigue agent of the present invention preferably contains the following amino acids in the following ratio. That is, the anti-fatigue agent preferably contains an amino acid composition comprising 6 kinds of amino acids in a predetermined ratio.

30-200 parts of proline;

60-140 parts of glycine;

25-260 parts by weight of alanine;

40-130 parts of lysine;

20-75 parts by weight of tryptophan;

15-40 parts of histidine.

In addition, the amino acid composition of the above composition preferably further contains the following amino acids in the following amount ratios. That is, the anti-fatigue agent preferably contains an amino acid composition comprising 8 kinds of amino acids in a predetermined ratio.

3-75 parts of tyrosine;

15-45 parts of arginine.

In addition, the amino acid composition of the above composition preferably further contains the following amino acids in the following amount ratios. That is, the anti-fatigue agent preferably contains an amino acid composition comprising 11 amino acids in a predetermined amount ratio.

30-55 parts by weight of valine;

35-60 parts by weight of leucine;

25-60 parts of isoleucine.

Further, the amino acid composition contained in the anti-fatigue agent of the present invention more preferably contains the following amino acids in the following amount ratios. That is, the anti-fatigue agent is more preferably an amino acid composition comprising 6 kinds of amino acids in a predetermined ratio.

30-200 parts of proline;

60-140 parts of glycine;

50-260 parts by weight of alanine;

50-130 parts of lysine;

30-75 parts by weight of tryptophan;

20-40 parts of histidine.

In addition, the amino acid composition of the above composition more preferably further contains the following amino acids in the following amount ratio. That is, the anti-fatigue agent is more preferably an amino acid composition comprising 8 kinds of amino acids in a predetermined ratio.

3-75 parts of tyrosine;

15-45 parts of arginine.

In addition, the amino acid composition of the above composition more preferably further contains the following amino acids in the following amount ratio. That is, the anti-fatigue agent is more preferably an amino acid composition comprising 11 kinds of amino acids in a predetermined ratio.

30-55 parts by weight of valine;

35-60 parts by weight of leucine;

25-60 parts of isoleucine.

Further, the amino acid composition contained in the anti-fatigue agent of the present invention preferably contains the following amino acids in the following amount ratios. That is, the anti-fatigue agent further preferably contains an amino acid composition comprising 6 kinds of amino acids in a predetermined amount ratio.

30-150 parts of proline;

60-110 parts of glycine;

50-220 parts by weight of alanine;

50-95 parts by weight of lysine;

30-65 parts by weight of tryptophan;

20-35 parts of histidine.

In addition, the amino acid composition of the above composition further preferably contains the following amino acids in the following amount ratios. That is, the anti-fatigue agent further preferably contains an amino acid composition comprising 8 kinds of amino acids in a predetermined amount ratio.

3-65 parts of tyrosine;

20-45 parts of arginine.

In addition, the amino acid composition of the above composition preferably further contains the following amino acids in the following amount ratios. That is, the anti-fatigue agent is more preferably an amino acid composition comprising 11 kinds of amino acids in a predetermined ratio.

30-50 parts by weight of valine;

40-60 parts by weight of leucine;

35-55 parts of isoleucine.

In addition, the amino acid composition contained in the anti-fatigue agent of the present invention preferably contains the following amino acids in the following amount ratios. That is, an anti-fatigue agent containing an amino acid composition comprising 6 kinds of amino acids in a predetermined amount ratio is particularly preferable.

30-100 parts of proline;

60-90 parts by weight of glycine;

100-220 parts by weight of alanine;

50-80 parts by weight of lysine;

30-55 parts of tryptophan;

20-30 parts of histidine.

In addition, in the amino acid composition of the above composition, it is particularly preferable to further contain the following amino acids in the following amount ratio. That is, an anti-fatigue agent containing an amino acid composition comprising 8 kinds of amino acids in a predetermined amount ratio is particularly preferable.

5-60 parts of tyrosine;

20-35 parts of arginine.

In addition, in the amino acid composition of the above composition, it is particularly preferable to further contain the following amino acids in the following amount ratio. That is, an anti-fatigue agent containing an amino acid composition comprising 11 kinds of amino acids in a predetermined amount ratio is particularly preferable.

30-45 parts by weight of valine;

45-60 parts by weight of leucine;

40-50 parts of isoleucine.

In the amino acid composition having the above-mentioned structure, tyrosine may be present in an amount of 3 to 30 parts by weight, and more preferably 5 to 25 parts by weight.

The amino acid composition as a main component of the anti-fatigue agent of the present invention is extremely safe and can be administered in a wide range. In general, the route of administration and various factors such as age, body weight, and symptoms of the subject animal including a human can be appropriately set in consideration. In the present invention, this is not limited, and for example, the amino acid composition is administered as an active ingredient preferably in the range of 0.01 to 8g/kg/day, more preferably 0.05 to 8g/kg/day, still more preferably 0.1 to 8g/kg/day, yet more preferably 0.3 to 8g/kg/day, particularly preferably 0.4 to 5g/kg/day, and most preferably 0.5 to 3 g/kg/day.

On the other hand, as shown by the results of human tests in the examples below, the anti-fatigue agent of the present invention can be administered, for example, preferably in a range of 0.01 to 0.3g/kg/day, more preferably in a range of 0.015 to 0.28g/kg/day, further preferably in a range of 0.02 to 0.25g/kg/day, further preferably in a range of 0.05 to 0.25g/kg/day, particularly preferably in a range of 0.05 to 0.2g/kg/day, and most preferably in a range of 0.05 to 0.15g/kg/day, in view of the use, effect, efficacy, production cost, and the like of the present invention, because the amino acid composition as an active ingredient can exert a sufficient effect at a lower concentration than ever before.

The composition of the present invention can be administered not only as an anti-fatigue agent in advance when prevention of fatigue is desired, but also as a fatigue recovery agent after the feeling of fatigue. The anti-fatigue agent of the present invention can be administered by either oral administration or non-oral administration (intramuscular, subcutaneous, intravenous, suppository, transdermal, etc.).

The anti-fatigue agent for oral administration may be administered, for example, 0.3g to 30g per day 1 to 3 times. When the agent is a liquid, it can be administered in the form of a 0.3 to 6.0 wt% solution 100 to 500ml per day 1 to 3 times. In the case of an injection to be administered intravenously, for example, 100 to 400ml, preferably 150 to 300ml of the solution may be administered in the form of a 0.3 to 6.0 wt% solution.

The anti-fatigue agent of the present invention was evaluated to have an anti-fatigue effect on both muscle fatigue and nerve fatigue using 2 evaluation systems, i.e., the measurement of the amount of movement during fatigue and the measurement of a blood biomarker during fatigue.

The evaluation by the action amount measurement is performed, for example, as follows: based on the following method for measuring the amount of action using mice, the amount of action of each of 2 groups, i.e., the administration group (administered with the amino acid composition) and the non-administration group (control group), was measured by applying a motor load to the 2 groups using a pedal rotator (pedal), and the like, and the relative amount of action of the administration group was calculated with the amount of action of the non-administration group as 100. When the amount of movement (relative value) of the administration group exceeds 100, preferably 110 or more, more preferably 120 or more, particularly preferably 190 or more, the effect of preventing muscular fatigue of the administered amino acid composition can be evaluated. The larger the numerical value of the amount of movement (relative value) of the administration group, the higher the effect of preventing muscle fatigue can be evaluated.

Evaluation of measurement using blood biomarkers is performed, for example, as follows: based on the following method for measuring a biomarker in blood using a mouse, 2 groups of an administration group (administration of an amino acid composition) and a non-administration group (control group) are subjected to a exercise load using a pedal rotor or the like, blood is collected after a certain period of time has elapsed, the concentration of the biomarker in blood of each of the 2 groups is measured, and the relative measurement concentration of the administration group is calculated assuming that the measurement concentration of the non-administration group is 100. When any blood biomarkers including cortisol, interferon γ (IFN- γ), and interleukin 10(IL-10) are used as an index, the effect of preventing nerve fatigue can be evaluated when the measured concentration (relative value) of the administration group is less than 100, preferably 96 or less.

In particular, when cortisol is used as an index, the effect of preventing nerve fatigue can be evaluated when the measured concentration (relative value) of the administration group is preferably 95 or less, more preferably 80 or less. When IFN- γ is used as an index, the effect of preventing nerve fatigue can be evaluated when the measured concentration (relative value) of the administration group is preferably 60 or less, more preferably 45 or less. When IL-10 is used as an index, the effect of preventing nerve fatigue can be evaluated when the measured concentration (relative value) of the administration group is preferably 70 or less, more preferably 50 or less.

The smaller the value of the measured concentration (relative value) of the administration group, the higher the effect of preventing the nerve fatigue can be evaluated.

The anti-fatigue agent of the present invention may contain, if necessary, methionine (preferably 0.3 to 0.7 mol%, more preferably 0.4 to 0.6 mol%), aspartic acid (preferably 0.1 to 0.3 mol%), taurine (preferably 3 mol% or less), phosphoethanolamine (preferably 2 mol% or less), cystine (preferably 0.5 mol% or less), β -alanine (preferably 1 mol% or less), γ -aminobutyric acid (preferably 0.5 mol% or less), ornithine or ethanolamine (preferably 3 mol% or less), ammonia (preferably 2 mol% or less), 1-methylhistidine (preferably 3 mol% or less), 3-methylhistidine (preferably 1 mol% or less), and the like, in addition to the amino acids.

The anti-fatigue agent of the present invention may contain, in addition to the above amino acids, components expected to have an effect of preventing fatigue, such as citric acid, carnitine, coenzyme Q10, dextrin (linear, branched, cyclic, etc.), vitamins, minerals, and the like, as necessary.

The amino acid used in the present invention is particularly preferably an L-amino acid, and each of the L-amino acids is preferably a single product and has high purity. For example, amino acids having a purity higher than that specified in "dietary supplement official " are used. In addition, these amino acids can also be used in the form of their physiologically tolerable salts.

The anti-fatigue agent of the present invention particularly preferably contains trehalose. Trehalose is present in many animals, plants and microorganisms in nature, for example in the nutrient exchange solutions of wasps and their larvae. In this case, it is considered that the synergistic effect of the amino acid composition of the present invention and trehalose is expected by using them together, and the effect of the anti-fatigue agent of the present invention in recovering from fatigue and preventing fatigue is improved. The mixing ratio of trehalose in the anti-fatigue agent of the present invention is not particularly limited, and may be suitably set according to the purpose or effect thereof, and is, for example, preferably 0.45 to 1.6: 0.5 to 5.0, more preferably 0.8 to 1.6: 1.0 to 4.0, and further preferably 1.0 to 1.6: 1.5 to 4.0 in terms of the mass ratio of the amino acid composition of the present invention to trehalose.

In the production of the amino acid composition of the present invention, commercially available amino acids may be mixed in the above-mentioned predetermined ratio. When the aqueous dispersion is used in the form of a solution, it may be dissolved in distilled water, ion-exchanged water, or the like. In general, the composition is prepared by uniformly mixing in powder form in advance, and is dissolved in distilled water, ion-exchanged water, or the like at the time of use. The temperature at which the composition of the present invention is produced or stored is not particularly limited, but it is preferably produced or stored at room temperature or lower.

The mode of administration of the anti-fatigue agent of the present invention is not particularly limited, and may be oral administration or non-oral administration according to a commonly used method. Specifically, the pharmaceutical composition can be administered in the form of, for example, tablets, powders, granules, capsules, syrups, troches (troche), inhalants, suppositories, injections, ointments, eye drops, nasal drops, ear drops, pastes, and lotions.

The above-mentioned preparations can be prepared by a conventional method by using a commonly used excipient, binder, lubricant, colorant, flavoring agent, deodorant, stabilizer, emulsifier, absorption enhancer, surfactant, pH adjuster, preservative, antioxidant, etc. as required, and blending the components generally used as raw materials of pharmaceutical preparations.

In the case of producing an oral preparation, for example, the amino acid composition of the present invention or a pharmacologically acceptable salt thereof is added with an excipient, and if necessary, a binder, a disintegrant, a lubricant, a coloring agent, a flavoring agent, a deodorant, or the like, and then the mixture is formulated into powder, fine granules, tablets, coated tablets, capsules, or the like by a conventional method.

Examples of the excipient include lactose, corn starch, white sugar, glucose, mannitol, sorbitol, crystalline cellulose, and silica. Examples of the binder include polyvinyl alcohol, polyvinyl ether, methyl cellulose, ethyl cellulose, gum arabic, tragacanth, gelatin, shellac, hydroxypropylmethyl cellulose, hydroxypropylcellulose, polyvinylpyrrolidone, polypropylene glycol polyoxyethylene block copolymer, and meglumine.

Examples of the disintegrating agent include starch, agar, gelatin powder, crystalline cellulose, calcium carbonate, sodium hydrogen carbonate, calcium citrate, dextrin, pectin, and calcium carboxymethylcellulose. Examples of the lubricant include magnesium stearate, talc, polyethylene glycol, silicon dioxide, and hardened vegetable oil.

The coloring agent may be a substance that is acceptable for addition to a medicine, and the flavoring and deodorizing agent may be cocoa powder, menthol, aromatic powder, peppermint oil, borneol, cinnamon powder, or the like. Examples of the flavoring deodorant include commonly used sweeteners, souring agents, and perfumes.

In the case of producing a liquid preparation such as syrup or an injection preparation, the amino acid composition of the present invention or a pharmacologically acceptable salt thereof is added with a pH adjuster, a dissolving agent, an isotonizing agent, and the like, and if necessary, a cosolvent, a stabilizer, and the like, and the preparation is carried out by a conventional method.

The anti-fatigue agent of the present invention contains the amino acid composition, and can exhibit sufficiently excellent effects as a pharmaceutical product, but can be prepared into a food by appropriately adjusting the concentration of the amino acid composition while taking into consideration the taste (flavor), and can also be prepared into a food for special use such as a food for special health use. The anti-fatigue agent of the present invention can be contained in a food composition and directly ingested as a functional food such as a nutritional functional food, thereby easily obtaining an effect of preventing fatigue.

Specifically, when the composition is used as a food composition, the anti-fatigue agent of the present invention may be added to various foods and beverages (e.g., milk, refreshing drink, fermented milk, yogurt, cheese, bread, biscuit, cracker, pizza, formula milk powder, liquid diet, food for patients, infant milk powder, milk powder for lactating women, and nutritional food) to be ingested. In addition, it can be used according to the conventional method in usual food compositions such as mixing with other foods or food ingredients. The state of the polymer may be any of the state of a generally used food or drink, for example, solid (powder, granule, etc.), gel, paste, liquid, or suspension.

When the composition is used as a food composition, other components are not particularly limited, and water, proteins, sugars, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, flavors (flavors), and the like contained in the food composition can be used as the components. Examples of the protein include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein concentrate, whey protein isolate, α -casein, β -casein, κ -casein, β -lactoglobulin, α -lactalbumin, lactoferrin, soy protein, egg protein, meat protein and other animal and vegetable proteins, hydrolysates thereof, butter, whey minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipids, lactose and other milk-derived components. Examples of the saccharides include various saccharides, processed starches (soluble starches other than dextrin, british starch, oxidized starches, starch esters, starch ethers, and the like), dietary fibers, and the like. Examples of the lipid include animal fats and oils such as lard and fish oil, their separated oils, hydrogenated oils and transesterified oils, and vegetable fats and oils such as palm oil, safflower oil, corn oil, rapeseed oil and coconut oil, their separated oils, hydrogenated oils and transesterified oils. Examples of the vitamins include vitamin a, carotenes, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, nicotinic acid, pantothenic acid, biotin, inositol, choline, folic acid, and the like; examples of the minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Examples of the organic acid include malic acid, citric acid, lactic acid, and tartaric acid. These components may be used in combination of 2 or more, or a synthetic product and/or a food containing a large amount of them may be used.

Examples

The present invention will be described in more detail below by referring to examples of the present invention, but the present invention is not limited to these specific examples, and various changes can be made without departing from the scope of the technical idea of the present invention.

Examples 1 to 4 and comparative example 1

Mice (male, 8-week-old, C57BL/6N, breeding environment: 23. + -. 3 ℃ C., 12-hour light and dark cycle) were divided into groups of 3 to 4 groups of test samples (examples) and control groups (comparative examples) as uniformly as possible. These mice were not fasted and were forcibly orally administered the anti-fatigue agent (test sample) or physiological saline (control) of the present invention. After 1 hour of administration, a movement load was applied by means of a pedal-rotator test (25m/min, 60 minutes).

Each sample was administered to the mice so as to achieve 500mg/kg body weight (suspension (5 wt%) was 10. mu.L/g body weight) in terms of amino acids. Here, the amino acid composition of the anti-fatigue agent of the present invention to be administered is shown in table 1 (the numerical values of the amino acid composition are shown in parts by weight).

(test 1: measurement of amount of movement in fatigue)

The amount of movement after the application of the movement load by the above-mentioned treadle was evaluated. After the end of the pedal test, the total amount of the spontaneous movement distance (moving distance) for 30 minutes under red lamp illumination was measured for the sample group (example) and the control group (comparative example). Then, the moving distance of the sample group is relatively represented as a moving amount with the moving distance of the control group as 100. That is, the larger the amount of action, the less muscular fatigue is considered. The measurement results are shown in Table 1.

(test 2: measurement of blood biomarkers in fatigue)

The biomarkers in blood after the exercise load was applied using the above-described treadmill were ranked. After the pedal rotator test is finished, blood is collected after a certain time, and the blood concentration of cortisol, interferon gamma (IFN-gamma) and interleukin 10(IL-10) is measured. Then, the respective numerical values of the sample groups were relatively expressed as immunological indicators in blood (blood biomarkers) with the respective numerical values of the control group as 100. That is, the smaller these values are, the lower the fatigue feeling and the less the nerve fatigue are considered. The measurement results are shown in table 1.

These evaluation methods were published in the 2005 society of biochemistry in japan (evolution: poor analysis of the site of immunity システム, biochemistry, 2005, Vol. 77, p.1056) according to the professor of biochemistry of the department of medicine, university of osaka city, and teaching of wellness for molecular pathology lecture (sa) and the like (rotation: fatigue における. the site of immunity システム. the rotation of rotation, 2005, Vol. 77, p.1056)

TABLE 1 amount of movement at fatigue and measurement results of blood biomarkers

Example No	Example 1	Example 2	Example 3	Example 4	Comparative example 1
						Pro	53.63	197.95	34.35	37.47	-
Gly	101.74	136.83	63.06	68.82	-
						Ala	256.56	52.05	166.64	181.82	-

Example No	Example 1	Example 2	Example 3	Example 4	Comparative example 1
						Lys	74.27	120.40	59.41	64.82	-
Trp	69.18	42.85	38.70	42.23	-
						His	32.85	38.14	21.06	23.00	-
Tyr	-	-	54.00	5.29	-
						Arg	-	-	23.64	25.82	-
Val	-	-	34.94	38.12	-
						Leu	-	-	49.76	54.29	-
Ile	-	-	42.65	46.53	-
						Kind of amino acid	6 kinds of	6 kinds of	11 kinds of	11 kinds of	0 species of
Amount of movement	110	119	285	298	100
						IFN-γ	56	38	51	38	100
Cortisol	79	76	50	74	100
						IL-10	96	61	48	65	100

In the evaluation of the amount of movement, it was found that the anti-fatigue agents of examples 1 to 4 of the present invention allowed further movement even after the application of a sports load by a treadmill, and had a higher effect of preventing muscle fatigue than comparative example 1. In particular, the amount of movement after exercise in examples 3 and 4 was found to be very high.

On the other hand, in the evaluation of blood biomarkers, it was found that all the values of cortisol, IFN-. gamma.and IL-10 were significantly reduced as compared with comparative example 1 by administering the anti-fatigue agents of examples 1 to 4 of the present invention. These compounds increase the fatigue feeling and increase the respective concentrations, and therefore can be evaluated as an index of the neural fatigue. That is, in examples 1 to 4 of the present invention, the effect of preventing the nerve fatigue is high as well as the effect of preventing the muscle fatigue. Moreover, these fatigue reducing (recovering) effects are particularly remarkable in examples 3 and 4.

[ example 5]

(test 3: fatigue resistance Effect verification test)

Next, an accelerated pulse examination, a blood examination, a urine examination, a saliva examination, and a physical examination (physical exam) were performed on an artificial subject, which were measured so as to be able to objectively evaluate muscle fatigue and nerve fatigue at the time of fatigue load without giving unnecessary psychological and physical pain to the subject, and the anti-fatigue effect of the anti-fatigue agent of the present invention was further verified.

(1) Subject of a human

The subjects were healthy adult male and female (12 male, 6 female, 18 total) judged by the test-responsible physician to be suitable for the trial from among the volunteers. The test was conducted under the gist of the declaration of Helsinki under the approval of the member under Investigation (IRB) at test .

(2) Test food

The test foods used in this test were 11 amino acid mixtures (hereinafter also referred to as "subject foods") and control foods (hereinafter also referred to as "placebo"). The nutrient composition and amino acid blend composition are shown in Table 2 and Table 3, respectively.

The subject food was hard capsules prepared from 11 amino acid mixtures in an amount of 200mg per capsule, and the placebo was hard capsules containing no 11 amino acid mixtures. In IRB, a functional test was performed to confirm that the subject food and placebo could not be identified by functional aspects such as flavor and aroma, or by packaging.

Table 2: test food nutrient composition (capsules each)

Heat-composition	Food subject	Placebo
			11 kinds of amino acid mixture (mg)	200	0
Crystalline cellulose (mg)	30	230
			Heat (kcal)	0.9	0
Protein (g)	0.2	0
			Lipid (g)	0	0
Carbohydrate (g)	0	0.2
			Sodium (mg)	0	0

Table 3: composition of 11 amino acid mixtures

(3) Test method

A. Design of experiments

The test was conducted in a double blind fashion, and was designed as a placebo-controlled 2-zone cross-control test. The test period was about 6 weeks in total, including a preceding observation period of 1 week, examination day, intake period of 1 week, and physical load day, which were performed 2 times at 4-week intervals (see fig. 1). The subjects ingested 5 test foods after each morning meal 1 week before the physical workload and immediately before the physical workload on the day of the loading. The subjects were instructed on the way of daily life such as diet and exercise before the test period was not changed.

B. Fatigue loading method by physical work

An Anaerobic working Threshold (AT) was measured before the previous observation period using a respiratory metabolism measurement system AE-300S (minito medical co., ltd.) and a load cell 75XL-IIME (Combi welless), and a physical working load on the day was obtained by using the load cell AT a load intensity of 80% of the heart rate AT the AT for 4 hours (1 hour × 4 group).

In addition, to supplement calories (glucose 75G) 10 minutes before the start of the load, please ingest 1 cigarette (225ml) of Toroeran G75(Ajinomoto Pharmaceuticals).

C. Test items

(a) Acceleration pulse examination

Acceleration pulses were measured by an acceleration pulse meter artemit C (U-medical) before the test food intake on the day of the intake period and the day of the load, 4 hours after the load was started and 4 hours after the recovery. The measurement time was 2 minutes and 1 time.

(b) Blood examination

Before the test food intake on the day of the intake period and the day of the load, and 30 minutes after the start of the load

Blood was collected (after a 10-second high-load test conducted 30 minutes after the start of loading), 4 hours after the start of loading, and 4 hours after recovery, and blood was examined. The test items were 37 items of blood biochemical test and concentrations of 41 amino acid-related substances including 20 amino acids.

(c) Urine examination

All urine at 4 hours of loading was collected on the day of loading and the concentration of vanillylmandelic acid, homovanillic acid and creatinine in the urine was determined.

(d) Saliva examination

Saliva was collected before test food intake on the day of loading, 2 hours after loading, 4 hours after loading and 4 hours after recovery, and the concentrations of amylase, cortisol, chromogranin a and total protein in saliva were determined.

(e) Physical examination

Blood pressure, pulse, body temperature and body weight were measured before intake of test food on the day of initiation of the intake period and the day of loading, 2 hours after initiation of loading, 4 hours after initiation of loading and 4 hours after recovery.

D. Test results

(a) Acceleration pulse examination

Among the measurement items, the measurement items for which significant differences were observed according to the standard that the risk ratio of t-test was 5% or less are as follows.

In the subject food group, compared to the placebo group, in absolute terms, it was found that the pulse rate was significantly decreased before the test food intake on the day of loading (0hr), Taz was significantly increased and e/a was significantly decreased 4 hours after the start of loading (4hr), the vascular aging deviation value was significantly decreased and Waveformindex I, d/a, the a-a interval variation coefficient, a-a interval standard deviation, and a-a interval variation amplitude were significantly increased 4 hours after recovery (8 hr). In addition, the change from the pre-ingestion value showed a significant decrease in pulse rate and a significant prolongation of Taa in terms of the change amount (0hr — before ingestion); in terms of the amount of change (4 hr-before ingestion), significant shortening of Taz was suppressed and Tab was significantly prolonged. In addition, it was found that, in terms of the change in load, a significant increase in the value of the aged vascular offset was suppressed, a significant decrease in the Waveform indexI was suppressed, and Taz was significantly shortened, as measured by the change amount (8hr-4 hr).

In the frequency analysis, the LF% -MEM on the day of onset of ingestion (before ingestion) was seen to be significantly increased in absolute value in the subject food group relative to the placebo group; LF-FFT, LF% -FFT significantly increased before intake of test food on the current day of loading (0 hr); LF% -MEM was significantly decreased, LF-FFT, HF-MEM were significantly increased 4 hours (4hr) after the start of the load; LF-FFT, total P-MEM, and total P-FFT increased significantly after 4 hours of recovery (8 hr). In addition, a significant decrease in LF% -MEM in the amount of change (4hr — before intake) from the amount before intake was observed. In terms of the change in load, it was found that the significant increase in LF% -MEM was suppressed and the LF% -FFT was significantly reduced in the amount of change (4hr to 0 hr).

Among the measurement items showing significant differences, the measurement results (absolute values) of d/a, a-a interval coefficient of variation, and LF% -MEM are shown in table 4.

Table 4: acceleration pulse measurement result

Mean. + -. standard deviation of the mean

With corresponding t-test: p < 0.05

In the above measurement results, d/a is an index reflecting the decrease in capillary resistance and is increased by the vasodilation action of the parasympathetic nerve, and therefore, it is considered that the food under test is a substance that suppresses the decrease in parasympathetic activity and regulates autonomic nerve function.

In addition, since the a-a interval coefficient of variation is an index reflecting parasympathetic nerve activity and is reduced by physical fatigue load, it is known that food under test is a substance that suppresses reduction in parasympathetic nerve activity due to fatigue and regulates autonomic nerve function.

In addition, LF% to MEM is an index reflecting sympathetic nerve activity, and since it is found that significant increase in LF% to MEM is suppressed, it is considered that food under test is a substance that suppresses increase in sympathetic nerve activity and regulates autonomic nerve function.

(b) Blood examination

Among the measurement items, the measurement items for which significant differences were observed according to the standard that the risk ratio of t-test was 5% or less are as follows.

In the subject food group, a significant decrease in uric acid on the day of initiation of ingestion (before ingestion) was seen in absolute values relative to the placebo group; triglyceride, sodium and chlorine were significantly decreased and free fatty acid was significantly increased 4 hours after the start of loading (4 hr). In addition, it was found that uric acid significantly increased and blood glucose significantly decreased in the amount of change (0 hr-before-intake), uric acid significantly increased and triglyceride significantly increased in the amount of change (4 hr-before-intake), and uric acid significantly increased in the amount of change (8 hr-before-intake). Furthermore, in terms of change under load, monocytes were significantly reduced in terms of change amount (4hr-0 hr); the alkaline phosphatase and calcium are remarkably reduced and the free carnitine is remarkably increased in terms of variation (8hr-0 hr); chlorine increased significantly by the amount of change (8hr-4 hr).

In the amino acid analysis, a significant increase in the proline/total amino acid ratio on the day of onset of intake (before intake) was seen in the subject food group relative to the placebo group in terms of absolute values; the tyrosine/total amino acid ratio and the aromatic amino acid/total amino acid ratio significantly increased 0.5hr after the start of loading (0.5 hr); the tyrosine/total amino acid ratio increased significantly 4 hours (4hr) after loading; after 4 hours (8hr) of recovery, the tyrosine/total amino acid ratio and the aromatic amino acid/total amino acid ratio were significantly increased. In addition, it was found that significant increases in the tryptophan/total amino acid ratio and the tryptophan/LNAA ratio were suppressed in the change amount (8hr — before ingestion) from the change amount before ingestion. In addition, in terms of the change under load, it was found that the ratio of lysine/total amino acid in terms of the amount of change (4hr to 0hr) and taurine were significantly increased; the significant decrease of cystine is inhibited by the variance (8hr-0 hr); the ratio of cystine and phenylalanine to total amino acid is significantly increased and taurine is significantly decreased in terms of the variation (8hr-4 hr).

Among the measurement items for which significant differences were observed, the measurement results (absolute values) of the concentrations of triglyceride and free fatty acid are shown in table 5.

Table 5: blood test measurement (Absolute value)

Item	Group of	Before ingestion	0hr	4hr	8hr
						TG(mg/dL)	Placebo group subject diet group	89±4793±53	94±5683±41	113±68]*95±57	79±5166±42
NEFA(mEq/L)	Placebo group subject diet group	345±105352±120	305±91349±127	1288±264]*1367±309	50±3453±36

Mean. + -. standard deviation of the mean

With corresponding t-test: p < 0.05

In the above measurement results, since a significant decrease in triglyceride and a significant increase in free fatty acid are observed, it is considered that the subject food promotes the decomposition of stored fat into free fatty acid (fat combustion) which becomes an energy source, and the energy supply can be adjusted according to the change in energy metabolism accompanying the increase in muscle load.

(c) Urine examination

The subject food group did not show significant difference from the placebo group according to the criteria that the risk rate according to the t-test was 5% or less.

(d) Saliva examination

Among the measurement items, the measurement items for which significant differences were observed according to the standard that the risk ratio of t-test was 5% or less are as follows.

In the test group, amylase was significantly decreased 4 hours (4hr) after the start of the loading, and amylase and salivary protein were significantly decreased 4 hours (8hr) after the recovery, in absolute terms, compared to the placebo group. In addition, in terms of change under load, it was found that significant increase in amylase was suppressed in terms of change amount (4hr to 0 hr); the significant increase of amylase and salivary protein is inhibited in terms of variation (8hr-0 hr).

Among the measurement items showing significant differences, the measurement results (absolute values and variation amounts) of Amylase (AMY) are shown in table 6.

Table 6: saliva examination and measurement results

Mean. + -. standard deviation of the mean

With corresponding t-test: p < 0.05

In the above measurement results, the amylase concentration in saliva is an index reflecting sympathetic nerve activity, and since a significant increase in amylase concentration is suppressed, it is considered that the subject food suppresses an increase in sympathetic nerve activity and regulates autonomic nerve function.

(e) Physical examination

Among the measurement items, the measurement items for which significant differences were observed according to the standard that the risk ratio of t-test was 5% or less are as follows.

In the subject food group, a significant increase in body temperature on the day of initiation of ingestion (before ingestion) was seen in absolute value relative to the placebo group. In addition, in terms of the change from the value before intake, a significant decrease in pulse rate and body temperature was observed in the change amount (0 hr-before intake), a significant decrease in blood pressure during the diastolic period was observed in the change amount (2 hr-before intake), and a significant increase in body temperature was observed in the change amount (8 hr-before intake). In addition, in terms of the change in load, a significant decrease in the diastolic blood pressure was observed in terms of the change amount (2hr to 0 hr).

From the results of tables 4 and 6, it is presumed that the anti-fatigue agent of example 5 of the present invention has effects of regulating autonomic nerve functions, alleviating mental load feeling of the subject, and preventing and recovering from nervous fatigue at the time of and after the load.

Further, from the results of table 5 above, it is presumed that the anti-fatigue agent of example 5 of the present invention promotes fat burning under load, increases the supply of free fatty acids as an energy source, and has the effect of preventing and recovering muscular fatigue of the subject.

Furthermore, in the above-described verification test of the anti-fatigue effect on the human subject, it is presumed that the amino acid composition of example 5 of the present invention has the effect of preventing and recovering muscle fatigue and nerve fatigue in humans as described above although the intake amount in the intake period is 1000mg/day (15.3 mg/kg/day calculated as 65.3kg of the average body weight of the subject) and extremely low, and therefore it is considered that a higher effect can be obtained by increasing the administration amount.

As described above, the anti-fatigue agent comprising the amino acid composition of the present invention can provide a high anti-fatigue effect of preventing both muscle fatigue and nerve fatigue. Further, since the composition is composed of a smaller number of types of amino acids than the conventional amino acid composition for the purpose of improving exercise performance, the composition requires a smaller number of types of raw materials for production, and has excellent industrial and economical effects. Therefore, the industrial value is high particularly in the field of functional amino acid compositions.

Claims (7)

1. An anti-fatigue agent comprising an amino acid composition comprising the following amino acids:

30-200 parts of proline;

60-140 parts of glycine;

50-260 parts by weight of alanine;

50-130 parts of lysine;

30-75 parts by weight of tryptophan;

20-40 parts of histidine.

2. The anti-fatigue agent according to claim 1, wherein the amino acid composition further comprises the following amino acids:

3-75 parts of tyrosine;

15-45 parts of arginine.

3. The anti-fatigue agent according to claim 2, wherein the amino acid composition further comprises the following amino acids:

30-55 parts by weight of valine;

35-60 parts by weight of leucine;

25-60 parts of isoleucine.

4. The anti-fatigue agent according to any one of claims 1 to 3, wherein the amino acid composition as an active ingredient is administered in the range of 0.01 to 8 g/kg/day.

5. The anti-fatigue agent according to any one of claims 1 to 4, wherein the anti-fatigue agent prevents both muscle fatigue and nerve fatigue at the same time.

6. The anti-fatigue agent according to claim 5, wherein muscle fatigue is evaluated by an action amount measurement, and nerve fatigue is evaluated by a blood biomarker measurement.

7. The anti-fatigue agent according to claim 6, wherein in the evaluation by an action amount measurement, when the action amount (relative value) of the non-administered group is 100 and the action amount (relative value) of the administered group is 110 or more, and in the evaluation by a blood biomarker measurement, when the measured concentration (relative value) of the non-administered group is 100 and the measured concentration (relative value) of the administered group is 96 or less, the effect of preventing muscle fatigue and nerve fatigue is evaluated to be exhibited.