JP2011148769A - Degradation inhibitor for branched chain amino acid - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new active ingredient having degradation-inhibiting effect of a branched-chain amino acid. <P>SOLUTION: There are provided a branched-chain amino acid degradation inhibitor containing lactoferrin as an active ingredient, a muscle fatigue inhibitor containing the branched-chain amino acid degradation inhibitor, and a muscle fatigue inhibitor further containing a branched-chain amino acid. There is provided a branched-chain amino acid degradation inhibitor which sufficiently inhibits degradation of the branched-chain amino acid, thereby capable of inhibiting muscle fatigue. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a branched chain amino acid degradation inhibitor.

  It is clear that branched chain amino acids (branched chain amino acids, BCAA) are involved in metabolic regulation under rapid exercise conditions and play an important role in the suppression of symptoms caused by muscle fatigue such as muscle damage and so-called muscle pain (Yoshiharu Shimomura, Descente Sports Science vol.28 p3-10 (Non-patent Document 1)). In other words, a certain amount of branched-chain amino acids are accumulated in muscle tissue, and the accumulated branched-chain amino acids are decomposed under rapid exercise conditions and used as energy sources (Yoshiharu Shimomura, Descente Sports Science) Vol.18 (Non-Patent Document 2)). In addition, when branched-chain amino acids are degraded under exercise conditions, the amount of branched-chain amino acids accumulated in muscle tissue decreases, which is estimated to contribute to the development of symptoms due to muscle fatigue. . As a method for suppressing such fatigue, a means for taking (supplementing) branched-chain amino acids from the outside before exercise is common.

  On the other hand, regarding lactoferrin, JP 2005-68060 A (Patent Document 1) discloses chronic fatigue syndrome such as vitamin B group complex, magnesium salt, calcium salt, carnitine, coenzyme Q10, sorghum extract, ginseng extract and the like. It is described that the therapeutic effect of the component is enhanced synergistically or additively in combination with a component effective for the treatment of the above. In addition, Natsume Ninagawa et al., Juntendo Medicine, Vol. 52, No. 4, 635, 2006 (Non-patent Document 3) suppresses an increase in blood lactate level and anemia by ingesting lactoferrin together with iron. Is observed.

JP 2005-68060 A

Shimomura Yoshiharu, Descente Sports Science vol. 28 p3-10 Effect of administration of branched chain amino acids on muscle damage caused by exercise Shimomura Yoshiharu, Descente Sports Science vol. 18 Kinetic and nutritional studies on proteins beneficial to muscle building through training Natsue Ninagawa et al., Effects of lactoferrin intake on blood chemistry markers indicating motor anemia in female long-distance runners, Juntendo Medicine 52, 4, 635, 2006

  With regard to the suppression of muscle pain, the effectiveness of the branched-chain amino acid intake from the outside alone is not sufficient, and a higher suppression effect is desired. On the other hand, the relationship between lactoferrin and the metabolism of branched chain amino acids and the effect of lactoferrin alone on the suppression of muscle pain have not been known so far. An object of the present invention is to provide a new active ingredient having an effect of inhibiting the degradation of branched chain amino acids.

  We have focused on the relationship between lactoferrin and the metabolism of branched-chain amino acids, and found that lactoferrin has the effect of suppressing the degradation of branched-chain amino acids in the process of intensive studies.

That is, the present invention provides the following [1] to [3].
[1] A branched-chain amino acid degradation inhibitor comprising lactoferrin as an active ingredient.
[2] A muscle fatigue inhibitor comprising the branched-chain amino acid degradation inhibitor according to [1].
[3] The muscle fatigue inhibitor according to [2], further comprising a branched chain amino acid.

  ADVANTAGE OF THE INVENTION According to this invention, the branched-chain amino acid degradation inhibitor which can fully suppress decomposition | disassembly of a branched-chain amino acid and can thereby suppress muscle fatigue is provided.

  The branched chain amino acid degradation inhibitor of the present invention contains lactoferrin as an active ingredient.

  Examples of lactoferrin include commercially available lactoferrin; mammals (eg, humans, cows, sheep, goats, horses, etc.) colostrum, transitional milk, regular milk, end milk, etc. or processed milk products of skim milk, whey, etc. And lactoferrin separated by a conventional method (for example, ion exchange chromatography); lactoferrin produced from plants (tomato, rice, tobacco); lactoferrin obtained by genetic recombination. Lactoferrin may be a commercially available product or can be prepared and used by a known method.

  As the lactoferrin, one of the above specific examples may be used alone, or two or more may be used in appropriate combination. In addition, as lactoferrin, the lactoferrin derived from a cow is preferable.

The branched chain amino acid degradation inhibitor of the present invention suppresses the degradation of branched chain amino acids.
The branched chain amino acid means an amino acid having a branched chain (branched side chain), and examples thereof include valine, leucine, and isoleucine. The optical isomerism of amino acids in the present invention is not particularly limited, but is usually L-type. The branched chain amino acid targeted by the branched chain amino acid degradation inhibitor of the present invention may be one of these, or two or more.

  Degradation of a branched chain amino acid means degradation of the chemical structure of the branched chain amino acid, and usually means degradation via the metabolic pathway of the branched chain amino acid in the body of an organism (in a tissue or in a cell). Usually, L-leucine is broken down into acetyl-CoA and acetoacetate, and L-valine and L-isoleucine are broken down into succinyl-CoA.

  Inhibition of the degradation of branched chain amino acids is usually to suppress the function of at least one of the various enzymes involved in the degradation pathway of branched chain amino acids in the organism, and the protein of the enzyme It is preferred to inhibit expression.

  The various enzymes are described in Kegg (Kyoto Encyclopedia of Genes and Genomes), and are specifically listed as follows.

  Aldehyde dehydrogenase 1 family, member B1 (aldehyde dehydrogenase 1 family, member B1), aldehyde dehydrogenase family 1, subfamily A7 (aldehyde dehydrogenase family 1, subfamily A7), aldehyde dehydrogenase 3 family, member A2 ), Aldehyde dehydrogenase 2 family (mitochondrial), aldehyde dehydrogenase 6 family member A1, aldehyde dehydrogenase 9 family, member A1 Aldehyde dehydrogenase such as

  Acyl-Coenzyme A dehydrogenase, C-2 to C-3 short chain, Acyl-Coenzyme A dehydrogenase, Acyl-Coenzyme A dehydrogenase , Short / branched chain), acyl-coenzyme A dehydrogenase, acyl-coenzyme A dehydrogenase, such as C-4 to C-12 straight chain (acyl-Coenzyme A dehydrogenase, C-4 to C-12 straight chain);

  Hydroxysteroid (17-β) dehydrogenase 4 (hydroxysteroid (17-beta) dehydrogenase 4), hydroxysteroid (17-β) dehydrogenase 10 (hydroxysteroid (17-beta) dehydrogenase 10), etc .;

  3-hydroxyisobutyrate dehydrogenase, dihydrolipoamide dehydrogenase, branched chain ketoacid dehydrogenase E1, alpha polypeptide, isovaleryl coenzyme A Dehydrogenases such as dehydrogenase (isovaleryl coenzyme A dehydrogenase), hydroxyacyl-Coenzyme A dehydrogenase;

  Acetyl-coenzyme A acetyltransferase 2 and other acetyl-coenzyme A acetyltransferase 1; acetyl-coenzyme A acetyltransferase 1;

  Acetyl-Coenzyme A acyltransferase 1 and acetyl-Coenzyme A acyltransferase 2; acetyl-Coenzyme A acyltransferase 2;

  Methylcrotonoyl-Coenzyme A carboxylase 2 (beta), methylcrotonoyl-Coenzyme A carboxylase 1 (alpha)), etc. -Coenzyme A carboxylase;

  Carboxylases such as propionyl-coenzyme A carboxylase, alpha polypeptide, propionyl coenzyme A carboxylase, beta polypeptide;

  Transferases such as 3-oxoacid-Coenzyme A transferase 1 (branched chain aminotransferase 2, mitochondrial), aldehyde oxidase 1;

  3-hydroxymethyl-3-methylglutaryl-Coenzyme A lyase, methylmalonyl-Coenzyme A mutase, 3-hydroxyisobutyl-Coenzyme A hydrolase (3-hydroxyisobutyryl-Coenzyme A hydrolase), 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (soluble), dihydrolipoamide branched Chain transacylase E2 (dihydrolipoamide branched chain transacylase E2), methylmalonyl CoA epimerase (methylmalonyl-Coenzyme A epimerase), enoyl coenzyme A hydratase, short chain, 1, mitochondrial (enoyl-Coenzyme A hydratase, short chain, 1 mitochondrial);

  Enoyl-coenzyme A, hydratase / 3-hydroxyacyl-coenzyme A dehydrogenase (enoyl-Coenzyme A, hydratase / 3-hydroxyacyl Coenzyme A dehydrogenase), AU RNA binding protein / enoyl-coenzyme A hydratase (AU RNA binding protein / enoyl-coenzyme) A hydratase), hydroxyacyl-coenzyme A dehydrogenase / 3-ketoacyl-coenzyme A thiolase / enoylcoenzyme A hydratase (trifunctional protein), β subunit (hydroxyacyl-Coenzyme A dehydrogenase / 3-ketoacyl-Coenzyme A thiolase / enoyl) -Coenzyme A hydratase (trifunctional protein), beta subunit), hydroxyacyl-coenzyme A dehydrogenase / 3-ketoacyl-coenzyme A thiolase / enoyl-coenzyme Hydratase (trifunctional protein), alpha subunit (hydroxyacyl-Coenzyme A dehydrogenase / 3-ketoacyl-Coenzyme A thiolase / enoyl-Coenzyme A hydratase (trifunctional protein), alpha subunit) enzyme complex, such as.

  The branched-chain amino acid degradation inhibitor of the present invention can express the activity of inhibiting the degradation of branched-chain amino acids by inhibiting the protein expression of one or more enzymes selected from the enzymes described above.

  In the present invention, suppressing decomposition means that the degree of decomposition of a substance in a living body is lower than usual. In the present invention, suppression of expression means that the degree of expression of a protein in a living body is lower than usual, and more specifically, the degree of expression of a protein gene is lower than usual. means.

  Confirmation that the branched chain amino acid degradation inhibitor of the present invention suppresses the degradation of branched chain amino acids can be performed, for example, as in the Examples. That is, the visceral fat differentiation medium (for example, a product of Primary Cell) with or without addition of lactoferrin is added to the preadipocytes prepared from rat mesenteric tissue and cultured. Thereafter, RNA is extracted from the cells and subjected to GeneChip analysis. GeneChip analysis uses a software such as GeneChip Data Analysis System GCOS (GeneChip Operating Software) to perform comparison analysis of samples with and without lactoferrin to determine the increase or decrease of the genes of enzyme proteins involved in the degradation of branched chain amino acids. Can be confirmed. In addition, when the sample is actually administered to a human or non-human mammal, the amount of degradation of the branched chain amino acid in the adipocytes before and after sample administration, the expression level of the enzyme involved in the degradation of the branched chain amino acid Changes, changes in the amount of branched chain amino acid degradation in adipocytes before and after sample administration may be confirmed.

  The object of the branched chain amino acid degradation inhibitor of the present invention is not particularly limited. Useful for humans or non-human vertebrates. Further, the health condition of the target human or non-human vertebrate is not particularly limited.

  The branched chain amino acid degradation inhibitor of the present invention may contain a pharmaceutically acceptable carrier in addition to lactoferrin. Examples of the pharmaceutically acceptable carrier include oily components, lubricants, excipients, binders, disintegrants and the like. Further, an appropriate amount of additives such as a sweetening agent, a sour agent, a flavoring agent, a coloring agent, and a pigment may be appropriately contained.

  Examples of the oil component include various fatty acid esters, hydrocarbons, higher fatty acids, higher alcohols and the like. Examples of the lubricant include magnesium stearate, stearic acid, sodium stearyl fumarate, sucrose fatty acid ester, polyethylene glycol (macrogol), talc and the like.

  As binders, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, gelatin, starch, dextrin, gum arabic, sodium alginate, tragacanth, purified gelatin, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, polyvinyl alcohol (partially saponified product), ethylcellulose, Examples include pullulan and polyethylene glycol (macrogol).

  Disintegrants include carboxymethylcellulose calcium (carmellose calcium), carboxymethylcellulose sodium (croscarmellose sodium), low-substituted hydroxypropylcellulose, carboxymethylcellulose, carboxymethylstarch sodium, cross-linked carboxymethylcellulose sodium (croscarmellose sodium) Examples thereof include powdered cellulose, cellulose or derivatives thereof, cross-linked polyvinyl pyrrolidone (crospopidone), starch, carboxymethyl starch, hydroxypropyl starch, agar and the like.

Excipients are exemplified as follows:
Polysaccharides such as crystalline cellulose, sodium alginate, xanthan gum;
Starches such as pregelatinized starch, hydroxypropyl starch, corn starch, potato starch and derivatives thereof;
Sugars and sugar alcohols such as sucrose, glucose, xylitol, erythritol, sorbitol, lactitol, trehalose, palatinose, palatinit (reduced palatinose), mannitol, maltitol, lactitol, lactose, fructose, powdered reduced maltose varicella;
Cellulose such as powdered cellulose, partially pregelatinized starch, ethylcellulose and derivatives thereof;
Light anhydrous silicic acid, titanium oxide, aluminum hydroxide gel, synthetic aluminum silicate, aluminum trisilicate, silicon dioxide, kaolin, cocoa butter, citric acid or its salt, stearic acid or its salt, calcium hydrogen phosphate, phosphoric acid Sodium hydride.

  Examples of the sweetener include aspartame, acesulfame potassium, saccharin sodium, sucralose, stevia, thaumatin and the like. Examples of the sour agent include citric acid, succinic acid, tartaric acid, malic acid, fumaric acid and the like. Examples of the flavoring agent include monoterpenes such as menthol, camphor, borneol, limonene, and various fragrances.

  The dosage form of the branched chain amino acid degradation inhibitor of the present invention is not particularly limited, and may be appropriately selected depending on the administration form. In the case of oral administration, tablets, capsules, granules, powders, fine granules, syrups, sustained-release tablets, tablets, chewable tablets, drop agents and the like can be mentioned.

  The production method of the branched chain amino acid degradation inhibitor is not particularly limited, and can be appropriately selected according to the dosage form. For example, when the dosage form is a tablet, a method of obtaining a tablet by mixing lactoferrin and optional components that can be blended as necessary, and then compressing the mixture to obtain a tablet, and further, the tablet obtained after compression molding as described above is enteric. A method of coating with components (method of enteric solvent) is mentioned, and the latter method is preferred. Examples of the enteric component include shellac, hydroxymethylcellulose phthalate, carboxymethylcellulose, aminoalkyl methacrylate copolymer, brewer's yeast cell wall (for example, trade name yeast wrap), tapioca starch, gelatin, pectin, etc. Among them, shellac is preferable. Whether or not it is an enteric solvent can be confirmed by the 14th revised Japanese Pharmacopoeia Disintegration Test Method.

  The branched chain amino acid degradation inhibitor of the present invention exhibits an excellent inhibitory effect on the degradation of branched chain amino acids. Therefore, the branched-chain amino chain degradation inhibitor of the present invention is effective in suppressing a phenomenon caused by degradation of a branched-chain amino acid, for example, muscle fatigue (for example, symptoms such as muscle damage and muscle pain). That is, when branched-chain amino acids are degraded under exercise conditions, the amount of branched-chain amino acids accumulated in muscle tissue decreases, which contributes to the development of symptoms due to muscle fatigue. If the decomposition of is suppressed, the accumulated amount is maintained, so that the effect of suppressing muscle fatigue is brought about. Therefore, the branched chain amino acid degradation inhibitor of the present invention can be used as a muscle fatigue inhibitor (that is, as a muscle fatigue inhibitor containing lactoferrin as an active ingredient). In addition to the above effects, lactoferrin is also known to have an effect of promoting fat metabolism (decomposition), that is, an obesity prevention and treatment effect. Therefore, muscle containing the branched-chain amino acid degradation inhibitor of the present invention By exercising while taking a fatigue inhibitor, it is expected that obesity can be effectively eliminated, and maintenance of health, prevention of lifestyle-related diseases, and treatment can be efficiently achieved.

  The blending amount of lactoferrin in the branched chain amino acid degradation inhibitor of the present invention can be appropriately determined according to the effect of inhibiting the degradation of branched chain amino acids. Generally, the blending concentration of lactoferrin is preferably 10% by mass or more, and more preferably 30% by mass or more.

  The dosage of the branched-chain amino acid degradation inhibitor of the present invention is the dosage form, dosage form, age of the human or non-human vertebrate to be administered, body weight, sex, exercise load when taken before exercise. Although depending on the conditions such as the above, the above-described effects can be exhibited, and the side effects can be appropriately determined within a range that can be tolerated. When the administration subject is an adult, it is usually preferable to administer an amount of 50 mg or more per day, more preferably 100 mg or more as the amount of lactoferrin. The upper limit is usually 5,000 mg or less. That is, it is preferable to administer an amount of 50 to 5,000 mg per day as the amount of lactoferrin. For example, if you want to take 300 mg per day with a tablet containing 100 mg of lactoferrin, you will take 3 tablets.

  The administration method can be appropriately determined depending on the blending concentration of lactoferrin, the dosage form, the age, weight, sex, and exercise load when taking the subject before exercise. For example, when the dosage form is a tablet, it is preferably taken with water or the like. The administration interval can be determined as appropriate, and may be any time before a meal, after a meal, or between meals.

  In addition to administration of the branched-chain amino acid degradation inhibitor of the present invention, administration of branched-chain amino acids or other branched-chain amino acid degradation inhibitors may be combined. When the administration of the branched chain amino acid degradation is combined with the administration of the branched chain amino acid degradation inhibitor of the present invention, it can be administered together with the branched chain amino acid degradation inhibitor of the present invention, or lactoferrin and the branched chain amino acid It is also possible to administer (as a single agent) as a muscle fatigue inhibitor containing

  By providing a branched-chain amino acid degradation inhibitor containing lactoferrin to humans or non-human vertebrates, muscle fatigue can be suppressed. Therefore, the branched chain amino acid degradation inhibitor of the present invention is also useful as a medicine, quasi-drug, and food.

  In addition to so-called health foods, foods include functional foods, functional health foods, foods for specified health use, functional nutritional foods, and dietary supplements.

Here, the intake time of the branched-chain amino acid degradation inhibiting food is not particularly limited, but is preferably taken before the start of exercise (sports, competition). Thereby, even at the time of exercise, the branched chain amino acids are not decomposed and the accumulation amount at the normal time (when not exercising) can be maintained, and muscle fatigue can be effectively suppressed. “Before starting exercise” means before the start of exercise, and is usually several hours before (for example, 30 minutes to 3 hours before).
There is no particular limitation on the exercise, and it can be applied regardless of the type and intensity of exercise, and the time of exercise, but it is preferably a strong exercise with muscle pain, that is, short-distance running or lack of exercise. It is a sudden and unusual movement of a person.

  In the case of food, it is preferable to display the effect of the present invention. The indication of the effect of the present invention includes an indication that it is used for inhibiting the degradation of branched chain amino acids; an indication that it is used for suppressing muscle fatigue, and the like.

  Thus, the food of the present invention suppresses the degradation of branched chain amino acids in the body by being administered and ingested to humans or non-human vertebrates. Therefore, the food of the present invention is useful as a food for suppressing muscle fatigue. In addition, since food can be taken daily, it is extremely useful as a preventive measure for muscle fatigue during exercise.

Example 1
Mesenteric adipose tissue was excised from SD rats (male, 12 weeks old), and preadipocytes were prepared. This was suspended in a visceral fat differentiation medium (Primary Cell) and seeded in a 24-well plate for cell culture. The cell seeding amount was 1 × 10 ⁷ cells / 24 well plate. At the start of culture, 0.3 mg / mL bovine lactoferrin (Wako Pure Chemical Industries) was added to each plate. The medium was changed once every two days during the culture period. In addition, it culture | cultivated similarly to the group which added lactoferrin as mentioned above (LF addition group) except not adding the lactoferrin as a control (non-addition group).

  Cells were collected 5 days after the addition of lactoferrin (5 days after the start of culture), total RNA was extracted and purified from the cells using RNeasy Mini Kit (QIAGEN), and the GeneChip step and Array scan were performed according to the protocol of Affymetrix. . For the Chip, a Rat Expression Array 230 2.0 was used. In scanning, image data was acquired using GeneChip 3000 Scanner.

  Using the GeneChip data analysis system GCOS (GeneChip Operating Software), the acquired array image data of each sample was confirmed. In addition, the gene expression level (Signal) was extracted as a numerical value using GeneChip data analysis software Expression Console. Further, using GCOS, a Comparison Analysis was performed between the LF added group and the non-added group, and the variation ratio (Signal Log Ratio = log2 (signal of the LF added group / signal of the non-added group)) was calculated, and the expression change (* 1 in Table 1-1 and Table 1-2). Tables 1-1 and 1-2 summarize the results of changes in gene expression on the fifth day of culture.

  As is clear from Table 1-1 and Table 1-2, in the LF added group, the expression of most of the enzyme groups related to branched chain amino acid degradation is significantly reduced compared to the non-added group. It was. This revealed that lactoferrin suppresses the degradation of branched chain amino acids.

Example 2
Formulation containing lactoferrin (lactoferrin 100 mg, crystalline cellulose 65 mg, lactose 60 mg, dextrin 45 mg, carboxymethylcellulose calcium 5 mg, sucrose fatty acid ester 5 mg, and Hihatsu fever) A plasticizer and a fluidizing agent were added to 5 mg of the water extract, tableted as usual, and 300 mg tablets coated with shellac were continuously taken, and the effect after taking was examined. The period of taking was about more than half a year (54% is more than one year). The dose was around 3 tablets per day.

  Regarding the ease of exercise, 12% (63) of the subjects answered that it became easier, and 1% (4) of the subjects answered that it became “tight”. In addition, among subjects who have a habit of exercising strongly (153 people), 16% (25 people) responded that they became easy, and 0% (0 people) answered that they became tight. )Met. Similarly, 15% (44) of subjects with a habit of moderate exercise (292) and 14% (61) of subjects (432) whose daily activities are walking became “easy”. And 0% (0) of the subjects answered that they became “tight”.

  Regarding physical activity increase / decrease, 15% (73) respondents replied “increased”, 6% (31 respondents) replied “decreased”, and the proportion of subjects who replied increased Was higher.

  From this, it was revealed that lactoferrin exhibits an effect of suppressing muscle fatigue. Moreover, based on the results of Example 1, it can be considered that lactoferrin inhibits the degradation of branched chain amino acids in the body, thereby suppressing muscle fatigue.

Claims (3)

  A branched-chain amino acid degradation inhibitor comprising lactoferrin as an active ingredient.   A muscle fatigue inhibitor comprising the branched-chain amino acid degradation inhibitor according to claim 1.   The muscle fatigue inhibitor according to claim 2, further comprising a branched chain amino acid.