JP7265322B2 - Suppressor of iron absorption inhibition by exercise and method for producing the same - Google Patents

Description

TECHNICAL FIELD The present invention relates to an inhibitor of exercise-induced iron absorption inhibition and a method for producing the same.

Exercise-induced anemia (also called sports anemia) is anemia that develops due to strenuous exercise. Exercise-induced anemia is a major problem, especially for athletes, because it affects athletic performance and health.
Exercise anemia is further classified into iron deficiency anemia, hemolytic anemia, and the like.
Of these, iron deficiency is the main cause of iron deficiency anemia. Also, the main cause of hemolytic anemia is the destruction of red blood cells due to compression of blood vessels in the soles of the feet during exercise.
Conventionally, iron preparations containing iron have been used to improve anemia. However, overdose of iron supplements carries the risk of side effects. That is, since humans do not have the function of excreting iron, iron may accumulate excessively in organs and cause dysfunction of the organs. In addition, gastrointestinal symptoms such as nausea (vomiting), vomiting, abdominal discomfort, abdominal pain, diarrhea, and constipation may occur.

Therefore, iron-free agents for improving exercise anemia have been proposed.
For example, Patent Document 1 describes a blood iron increasing agent for high-load exercisers containing catechins as an active ingredient. Patent Literature 1 describes that as an effect of this agent, ingestion of catechins during high-intensity exercise such as long-distance runners increases iron in the blood and prevents anemia. there is
Further, in Patent Document 2, the ability to remove risk factors during exercise (for example, the ability to prevent sports anemia) is characterized by containing an n-3 highly unsaturated fatty acid (for example, eicosapentaenoic acid) as an active ingredient. Compositions having are described.

JP 2017-43548 A JP-A-11-239464

Exercise anemia may occur even in athletes who are properly managing their diet and are taking enough iron. It is known that in some cases no improvement is observed.
Thus, at least some types of exercise anemia cannot be improved by iron supplementation. The reason for this is thought to be that the utilization of iron in vivo is inhibited.
An object of the present invention is to provide an agent capable of suppressing inhibition of iron utilization in vivo and having no risk of side effects when ingested.

As a result of intensive studies to solve the above problems, the present inventors have found that the hydrolyzate of milk-derived protein suppresses the inhibition of the utilization of iron in vivo without causing the side effects of iron preparations. I found that it can be done, and completed the present invention.

The present invention provides the following [1] to [7].
[1] A suppressor for inhibiting iron absorption due to exercise, comprising a milk-derived protein hydrolyzate as an active ingredient.
[2] The inhibitor of iron absorption inhibition due to exercise according to [1] above, which is an agent to be taken before exercise.
[3] The inhibitor of iron absorption inhibition by exercise according to [1] or [2] above, wherein the inhibitor of iron absorption inhibition by exercise is an iron-free agent.
[4] The inhibitor of iron absorption inhibition due to exercise is intended to be taken by the consumer in an amount of 0.01 to 5 g/day per kg of body weight as the hydrolyzate of the milk-derived protein. The inhibitor of iron absorption inhibition according to any one of [1] to [3] above.
[5] The inhibitor of iron absorption inhibition according to any one of [1] to [4] above, wherein the inhibitor of iron absorption inhibition due to exercise is a food or drink.
[6] A method for producing the inhibitor of iron absorption inhibition according to any one of [1] to [5] above, wherein the milk-derived protein is hydrolyzed using a proteolytic enzyme, A method for producing an agent for inhibiting iron absorption due to exercise, comprising a hydrolysis step of obtaining a hydrolyzate of the milk-derived protein.
[7] The method for producing an inhibitor of exercise-induced iron absorption inhibition according to [6] above, wherein the proteolytic enzyme is a microbial protease.

The present invention also provides the following [8] to [14].
[8] Use of a milk-derived protein hydrolyzate in the production of an inhibitor of iron absorption inhibition by exercise.
[9] The use according to [8] above, wherein the inhibitor of iron absorption inhibition due to exercise is an agent to be taken before exercise.
[10] The use according to [8] or [9] above, wherein the inhibitor of iron absorption inhibition by exercise is an iron-free agent.
[11] The inhibitor of iron absorption inhibition by exercise is intended to be ingested by the intake person in an amount of 0.01 to 5 g/day per 1 kg of body weight as the amount of the hydrolyzate of the milk-derived protein. The use according to any one of [8] to [10] above.
[12] The use according to any one of [8] to [11] above, wherein the inhibitor of iron absorption inhibition by exercise is a food or drink.
[13] A milk-derived protein hydrolyzate for use in the production of an inhibitor of exercise-induced iron absorption inhibition.
[14] A method for suppressing exercise-induced inhibition of iron absorption, which comprises the step of allowing a subject to ingest a milk-derived protein hydrolyzate.

According to the inhibitor of iron absorption inhibition by exercise of the present invention (herein, also referred to as the agent of the present invention), inhibition of iron utilization in vivo can be suppressed without causing side effects. can.
The agent of the present invention is effective for athletes with exercise anemia despite adequate iron intake through diet control, and for athletes with iron deficiency anemia despite taking iron preparations. It can be expected to improve anemia in those who cannot

The inhibitor of iron absorption inhibition by exercise of the present invention contains a milk-derived protein hydrolyzate as an active ingredient.
Examples of milk include animal milk such as cow's milk, sheep's milk and goat's milk.
Examples of milk-derived proteins include milk proteins and glycoproteins contained in exocrine fluids such as milk.
Milk proteins include casein and whey protein (also called whey protein).
Examples of whey proteins include α-lactalbumin and β-lactoglobulin.
Glycoproteins contained in exocrine fluids such as milk include lactoferrin.
Examples of milk-derived protein content include whey protein isolate (WPI; also referred to as whey protein isolate), whey protein concentrate (WPC), milk protein concentrate (MPC; including casein and whey protein). products), α-lactalbumin, β-lactoglobulin, bovine lactoferrin (lactoferrin derived from cattle), and the like.

A milk-derived protein hydrolyzate can be obtained by hydrolyzing a milk-derived protein using a proteolytic enzyme.
Examples of proteolytic enzymes include microbial proteases (microbial-derived proteases), plant proteases, mammalian proteases, fungal proteases, and the like.
Among them, microbial protease is preferably used in the present invention because it can efficiently produce a peptide having a preferred molecular weight, which will be described later.
In the present invention, a commercially available product can be used as the microbial protease.

Hydrolysates of milk-derived proteins contain peptides.
The number average molecular weight (Mn) of the peptide (hydrolyzate) is preferably 400-800, more preferably 450-700, particularly preferably 500-700. When the value is within the preferred range, the effect of the present invention (inhibition of iron utilization in vivo) can be further enhanced.
The weight average molecular weight (Mw) of the peptide (hydrolyzate) is preferably 500 to 1,200, more preferably 550 to 1,150, still more preferably 600 to 1,100, particularly preferably 600 to 900. . When the value is within the preferred range, the effect of the present invention (inhibition of iron utilization in vivo) can be further enhanced.
The weight average molecular weight to number average molecular weight ratio (Mw/Mn) of the peptide is preferably 1.0 to 1.4, more preferably 1.0 to 1.3, and particularly preferably 1.1 to 1.3. is. When this value is within the preferred range, the molecular weight distribution is narrow, so that the effect of the present invention can be enhanced when the same amount of the agent of the present invention is used, compared to when the molecular weight distribution is wide.
The number average molecular weight and weight average molecular weight of the peptide can be measured using, for example, size exclusion chromatography.

The agent of the present invention is preferably an iron (Fe)-free agent. When iron is not included, various symptoms (eg, nausea, vomiting, abdominal discomfort, etc.) due to iron content do not occur, and excessive accumulation of iron does not cause dysfunction of organs.
The agent of the present invention can be used for food and drink.
The agent of the present invention can contain various optional ingredients in addition to the milk-derived protein hydrolyzate, depending on its use. Examples of optional ingredients include water, proteins, amino acids, carbohydrates, lipids, vitamins, minerals, fruit juices, flavors and the like.
Examples of amino acids include arginine, leucine, isoleucine, and valine. Examples of vitamins include vitamin A, vitamin B group, vitamin C, vitamin D group, vitamin E, carotenes and the like. Examples of minerals include calcium, potassium, magnesium, sodium, zinc and the like.

The amount of the agent of the present invention to be used is preferably 0.001 to 2 g/day, more preferably 0.01 to 1 g/day, and more preferably 0.01 to 1 g/day, per 1 kg of body weight of the intake person. An amount of 0.015 to 0.5 g/day is preferred, and an amount of 0.02 to 0.2 g/day is particularly preferred.
When the amount is 0.001 g/day or more, the effect of the present invention (inhibition of iron utilization in vivo) can be further enhanced. When the amount is 2 g/day or less, it is possible to reduce the physical and mental burden on the ingestion person and the cost.

EXAMPLES The present invention will be described below with reference to Examples, but the present invention is not limited to these Examples.

[Whey peptide]
As the milk protein hydrolyzate, a product prepared by treating WPI (whey protein isolate) with a microorganism-derived protease (hereinafter referred to as whey peptide) was used. The weight average molecular weight was 717 and the number average molecular weight was 595 as measured by size exclusion chromatography.
As the size exclusion chromatography, "TSKgel G2000SWXL" (product name; manufactured by Tosoh Corporation) was used as the column, and 55% by weight water/45% by weight acetonitrile/0.1% by weight trifluoroacetic acid was used as the developing solvent. . The flow rate was set at 0.5 mL/min, and the temperature of the column oven was set at 30°C. Detection was performed by measuring absorbance at a wavelength of 214 nm. As molecular weight markers, cytochrome c (molecular weight: 12,400), adrenocorticotropic hormone (1-24) (molecular weight: 2,933), oxytocin (molecular weight: 1,007), and glutathione (molecular weight: 307) were used. board.

[Grouping of rats]
Forty rats (inbred rats, “DA rats”, male, 7 weeks old) were purchased from Japan SLC, Inc. and acclimatized for one week or longer. During the acclimatization period, standard diet MF was used as the diet. After acclimation, 4 animals with low body weight were excluded and divided into 4 groups of 9 animals per group.
The composition of these four groups (presence or absence of exercise load and administration) is as follows.
Group 1: No exercise load, administer distilled water Group 2: Exercise load, administer distilled water Group 3: Exercise load, administer whey peptides before exercise Group 4: Exercise load, administer whey peptides after exercise

[Rat preparatory exercise]
The diet was switched to the standard diet AIN-93G, and the rats (groups 1 to 4) were made to perform treadmill exercise in the morning for 5 consecutive days. The conditions for this treadmill exercise were set at a gradient of 0 degrees, a speed of 20 m/min, and a running time of 20 minutes. This was followed by a rest period of 4 days. Rats were fasted overnight starting in the evening on the last day of the rest period.

[Imposition of exercise load to rats (groups 2 to 4) and administration of iron]
In the morning of the day following the last day of the rest period after the preliminary exercise, the rats with "exercise load" (groups 2 to 4) were subjected to treadmill exercise under more severe conditions than the preliminary exercise described above. added load. The conditions for this treadmill exercise were set at a gradient of 5 degrees, a speed of 22 m/min, and a running time of 120 minutes.
At this time, 30 minutes before the exercise load and immediately after the end of the exercise load for the rats in Group 2 (with exercise load, distilled water administered) (more specifically, after blood collection as described later; hereinafter the same ), distilled water was administered orally in a volume of 10 milliliters per kg of body weight.
A mixture of whey peptides and distilled water was administered to the rats in group 3 (exercise load, whey peptides administered before exercise) 30 minutes before exercise load, and the amount of whey peptides was 1 g per 1 kg of body weight and It was administered orally in a volume of 10 milliliters of distilled water. In addition, distilled water was orally administered in an amount of 10 ml per 1 kg body weight immediately after the end of the exercise load.
Distilled water was orally administered to the rats in group 4 (exercise load, whey peptide administered after exercise) in an amount of 10 ml/kg body weight 30 minutes before the exercise load. Immediately after the end of the exercise load, a mixture of whey peptides and distilled water was orally administered in an amount of 1 g of whey peptides and 10 ml of distilled water per 1 kg of body weight.
Next, the rats (groups 2 to 4) were treated with an aqueous solution of iron sulfate (FeSO ₄ ) (concentration: 0.498 g/liter) 4 hours after the end of the exercise load. It was administered orally at a dose of 0.5 mg per dose.

[Iron administration to rats (group 1)]
After the rest period after the above preliminary exercise, the experiment was conducted in the same manner as the experiment of the rats of Group 2 (exercise load, distilled water administered) except that the exercise load was not applied.
Specifically, the rats in Group 1 (no exercise load, administered with distilled water) were allowed to rest without any exercise load while the rats in Group 2 were under exercise load. Distilled water was orally administered in an amount of 10 milliliters per kg of body weight to the rats in the group 30 minutes before the exercise load and immediately after the end of the exercise load.
Then, to the rats (group 1), 4 hours after the end of the exercise load to the rats of group 2, an aqueous ferrous sulfate solution was orally administered in the same manner as the rats of group 2.

[Chronological blood sampling]
For rats (groups 1 to 4), (a) immediately before oral administration of iron (iron sulfate) (“0 hr” in Table 1), (b) 1 hour after oral administration of iron (iron sulfate) ("1 hr" in Table 1), (c) 3 hours after oral administration of iron (iron sulfate) ("3 hr" in Table 1), a total of 3 times, chronological blood sampling from the jugular vein under isoflurane anesthesia. gone. After the last (3rd) time-course bleed, the rat was euthanized by exsanguination by drawing blood from the abdominal vena cava under isoflurane anesthesia.
[Measurement of plasma iron concentration in collected blood]
Plasma iron concentration of the collected blood of rats (groups 1 to 4) was measured by "metalloassay iron measurement LS (nitroso PSAP method)" (metallogenics kit).
Table 1 shows the measurement results (mean value, standard error) of plasma iron concentration.

[Statistical analysis]
Statistical analysis was performed using the statistical analysis software "Excel Statistics" (product name) (manufacturer: Social Information Service Co., Ltd.). Comparisons between rats and two groups of rats were performed by Dunnett's test.
As a result, one hour after oral administration of iron (1 hr), the rats in group 2 (with exercise load, administered distilled water) compared to the rats in group 1 (without exercise load, administered distilled water) showed significantly lower plasma iron concentrations at the 5% significance level (p<0.05). From this, it was confirmed that the iron absorption after the exercise load is inhibited by the exercise load.
In addition, at 1 hour after oral administration of iron (1 hr), the rats of group 3 (with exercise load, administration of whey peptide before exercise) rats of group 2 (with exercise load, administration of distilled water) In contrast, the amount of change in plasma iron concentration showed a significantly higher value at the significance level of 5% (p<0.05). From this, it was confirmed that the ingestion of whey peptides before exercise suppresses the inhibition of iron absorption after exercise.
One hour after oral administration of iron (1 hr), the rats in group 4 (with exercise load, whey peptide administered after exercise) and the rats in group 2 (with exercise load, administered distilled water) In contrast, plasma iron concentrations were elevated.
From the above, it was found that the agent of the present invention can suppress exercise-induced inhibition of iron absorption.

Claims (7)

An inhibitor of iron absorption inhibition due to exercise containing a milk-derived protein hydrolyzate as an active ingredient,
The milk-derived protein hydrolyzate is a whey protein isolate hydrolyzate having a number average molecular weight of 400 to 800 and a weight average molecular weight of 500 to 1,200. A inhibitor of exercise-induced iron absorption inhibition , which is a peptide having a ratio of 1.0 to 1.4 . 2. The inhibitor of iron absorption inhibition due to exercise according to claim 1, wherein the inhibitor of iron absorption inhibition due to exercise is an agent to be taken before exercise. 3. The inhibitor of iron absorption inhibition by exercise according to claim 1 or 2, wherein the inhibitor of iron absorption inhibition by exercise is an iron-free agent. The inhibitor of iron absorption inhibition due to exercise is to be taken by the consumer in an amount of 0.001 to 2 g/day per 1 kg of body weight as the amount of the milk-derived protein hydrolyzate. Item 4. The inhibitor of exercise-induced iron absorption inhibition according to any one of items 1 to 3. The inhibitor of iron absorption inhibition due to exercise according to any one of claims 1 to 4, wherein the inhibitor of iron absorption inhibition due to exercise is food or drink or a pharmaceutical. A method for producing the inhibitor of exercise-induced iron absorption inhibition according to any one of claims 1 to 5 , wherein the whey protein isolate is hydrolyzed using a proteolytic enzyme to produce a number of characterized by including a hydrolysis step of obtaining a peptide having an average molecular weight of 400 to 800, a weight average molecular weight of 500 to 1,200, and a weight average molecular weight to number average molecular weight ratio of 1.0 to 1.4. A method for producing an inhibitor of exercise-induced iron absorption inhibition. 7. The method for producing an inhibitor of exercise-induced iron absorption inhibition according to claim 6 , wherein the proteolytic enzyme is a microbial protease.