JP7429604B2 - Composition for suppressing TGF-β - Google Patents

Description

Transforming Growth Factor-β (TGF-β) is a type of cytokine that was initially discovered as a factor that promotes the transformation of fibroblasts. It has become clear that they are involved in a variety of biological phenomena by regulating the proliferation, motility, and differentiation of cell types. Furthermore, there are many known cases in which disruption of TGF-β signaling regulation leads to diseases (cancer, immune abnormalities, fibrosis, osteochondral diseases, vascular lesions, etc.), and the development of drugs that suppress the effects of TGF-β. It is also popular (Non-patent Document 1).

In addition, physical activities such as daily living activities and exercise are lifestyle habits that are essential for maintaining health. Lack of physical activity is said to be a risk factor for obesity, the development of lifestyle-related diseases, and a decline in independence and frailty in the elderly, so it is important to exercise for a certain amount of time and establish appropriate daily activities and exercise habits. This is very important. It is known that TGF-β causes the "central fatigue" that is felt long before reaching physical limits during exercise. For example, in Non-Patent Document 2, it was reported that cessation of spontaneous locomotor behavior was caused in mice administered post-exercise rat brain spinal fluid, and that the active substance that causes the cessation of movement in mice was It has been reported that it was TGF-β. Since 80% of the total amount of spinal fluid in the brain is derived from serum/plasma, the composition is very similar to that of serum/plasma (Non-Patent Document 3). It is also known that TGF-β in plasma increases during exercise stress (Non-Patent Document 4). Based on these facts, it is thought that central fatigue can be expected to be improved or prevented by suppressing the increase in TGF-β in plasma during exercise.

For example, Patent Document 1 discloses a TGF-β inhibitor containing theanine as an active ingredient. Patent Document 2 discloses a TGF-β2 inhibitor containing lactic acid bacteria (Lactococcus lactis subsp. cremoris H-61) as an active ingredient.
In the prior art, compositions for suppressing TGF-β have been exemplified, but the TGF-β suppressing effect of wheat protein hydrolysates has not been clarified.

As an effect of glutamine-containing peptides, it has been reported that they reduce delayed muscle damage after exercise loads in athletes such as long-distance running, soccer, and throwing (Non-Patent Documents 5 to 7). In addition, it has been reported that NK cell activity increases in healthy adults who do not have a habit of exercising (Non-Patent Document 8), and that muscle mass is maintained in middle-aged and elderly people when combined with body weight exercise (Non-Patent Document 9). However, the TGF-β inhibitory effect has not been clarified.

Furthermore, Patent Document 3 reports that, as an effect of a wheat-derived glutamine-containing peptide, it improves the feeling of fatigue the day after exercise and muscle pain 3 days after exercise.
However, Patent Document 3 does not relate to alleviation of central fatigue felt during exercise, and furthermore, the TGF-β suppressing effect is neither described nor suggested.

Patent No. 2006-63003 Patent No. 2015-145356 Japanese Patent Application Publication No. 2005-97162

“New developments in TGF-β signaling research” Keiji Miyazawa, Medical History, Vol.234(No.10), p885-889, 2010 “Exercise/Nutrition and Central Fatigue Generation Mechanism - Central Fatigue Generation and Metabolic Regulation by TGF-β”, Kazuo Inoue et al., Physical Fitness Science, 55(2), p279-286, 2006 “Cerebrospinal fluid omics” Masato Ishihara, Fluid Management Renaissance, July 2015 issue (Vol.5 No.3) p68-75 “Analysis of digestive and immune pathologies associated with exercise and development of preventive drinking water,” Masayasu Inoue et al., Descente Sports Science, Vol. 25, p88-93 Koikawa N et al. Delayed-onset muscle injury and its modification by wheat gluten hydrolysate. Nutrition 2009; 25: p493-498. Aoki K, et al. Post-training of wheat gluten hydrolysate suppresses the delayed onset consumption of muscle injury in soccer players. Experimental and Therapeutic Medicine 2012;3:p969-972. Yuta Takanashi et al. Does wheat gluten hydrolyzate (WGH) reduce muscle damage during weight training in throwing athletes? Journal of the Japanese Society of Clinical Sports Medicine 2012; 20(1): 66-71. Horiguchi N, et al. Effect of Wheat Gluten Hydrolysate on the Immune System in Healthy Human Subjects. Biosci Biotechnol Biochem. 2005;69(12):p2445-2449. Yuki Niimura et al. Effects of combined weight-bearing exercise and supplements containing wheat gluten hydrolyzate on lower limb strength and muscle mass in middle-aged and elderly people. Physical Fitness Science 2012; 61(6): 636(42-2-C-3, p188).

An object of the present invention is to provide a novel composition for suppressing TGF-β, which contains a wheat protein hydrolyzate as an active ingredient.

As a result of studies to solve the above-mentioned problems, the present inventors discovered that a hydrolyzate of wheat protein has a TGF-β suppressing effect, leading to the completion of the present invention.
Specifically, the hydrolyzate of wheat protein preferably contains a glutamine-containing peptide, particularly preferably the content of the glutamine-containing peptide is 65-90% by mass, and the proportion of glutamine and glutamic acid in the peptide is 30-90% by mass. The present invention provides a composition for suppressing TGF-β, in which the wheat protein hydrolyzate contains 30 to 70% by mass of peptides having a molecular weight of 5000 or more. The present invention also provides a TGF-β inhibiting food containing the above composition.

The composition of the present invention has the effect of reducing blood TGF-β during exercise stress. The composition of the present invention can prevent or improve central fatigue associated with exercise based on the TGF-β suppressing effect. Furthermore, preventive or therapeutic effects based on the TGF-β suppressing action can be expected for various diseases caused by TGF-β (immune disorders, fibrosis, osteochondral diseases, vascular lesions, etc.).

Hereinafter, preferred embodiments of the present invention will be specifically described.
The wheat protein hydrolyzate used in the present invention is obtained by hydrolyzing a high-molecular wheat-derived protein into a relatively small molecular weight peptide, which has a high TGF-β inhibitory effect. It is. For example, by ingesting the composition of the present invention before or during exercise, a reduction in subjective exercise load intensity can be expected.

Any available wheat can be used as the wheat. Suitable examples include bread wheat, durum wheat, club wheat, spelt wheat, eel wheat, etc. belonging to the genus Triticum of the family Poaceae, and wheat wheat and wheat wheat belonging to the genus Aegilops of the family Poaceae.

The wheat protein used for hydrolysis mainly refers to wheat-derived gluten (wheat gluten), but it also contains other proteins known to be contained in wheat, such as albumin, globulin, glutenin, and gliadin. Furthermore, impurities such as starch and fibers may be unavoidably contained.

In addition, in the present invention, wheat proteins that have been previously subjected to chemical treatment or biological treatment using enzymes, etc. to reduce the molecular weight of wheat-derived gluten, and those that have increased affinity with proteases are also used. can. Furthermore, unrefined wheat protein, unrefined wheat gluten, etc. can also be used. Wheat protein is obtained as a by-product when starch is purified from wheat, but it may be used as is, and by doing so, wheat protein, a by-product that has not been used in the past, can be effectively used.

The wheat protein hydrolyzate may be any product obtained by hydrolyzing the above-mentioned wheat protein, and the hydrolysis may be carried out by an appropriate known method, such as a method of hydrolyzing using an acid, or a method of hydrolyzing protein. This can be carried out by applying a method of hydrolysis using a degrading enzyme (protease).

As a method for hydrolyzing wheat protein using an acid, a conventional method can be adopted. For example, examples of acids that can be used include mineral acids such as sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, and sulfurous acid; organic acids such as oxalic acid, citric acid, acetic acid, and formic acid. The procedure for hydrolyzing wheat protein using such an acid is to add an acid to an aqueous medium containing wheat protein so that the acid normality is in the range of 0.01 to 2 N, and then lower the temperature of the aqueous medium. The procedure includes hydrolysis at 50 to 100°C for 10 minutes to 6 hours. If it is desired to stop hydrolysis, it can be stopped by adding a base to neutralize it.

When hydrolyzing using an acid, the concentration of wheat protein in the aqueous medium needs to be adjusted appropriately depending on the type and normality of the acid, but it is usually adjusted to a range of 1.0 to 80% by mass. It is better to do so.

Examples of the method of hydrolyzing wheat protein using a protein hydrolase include a method of hydrolyzing wheat gluten using protease, or protease and amylase (Japanese Patent No. 2019439 and Japanese Patent No. 2985193). (see official bulletin). As the protease in this case, various proteases such as pepsin, acidic protease derived from Hirotake, acidic protease derived from Aspergillus, papain, bromelain, thermolysin, trypsin, chymotrypsin, pronase, subtilisin, esperase, etc. can be used. . A single type of protease may be used, or a combination of multiple types, such as a combination of acidic protease and neutral protease, may be used, or the treatment may be performed multiple times.

When wheat gluten is hydrolyzed with protease, when amylase is used in combination, impurities such as starch and fiber contained in wheat gluten are decomposed and removed, resulting in a high yield of hydrolyzate containing highly purified peptide components. (See Japanese Patent No. 2985193). Wheat gluten is insoluble in water and has poor dispersibility in water, but a hydrolyzate of wheat gluten has a low molecular weight and therefore has excellent dispersibility in water.

The wheat protein hydrolyzate thus obtained may be used as it is as an active ingredient in the present invention. Therefore, the wheat protein hydrolyzate used in the present invention may contain, in addition to the peptide component, other components and enzymes derived from wheat protein and wheat gluten that are unavoidably included in the manufacturing process. .

However, in consideration of ease of handling, storage stability, etc., it is preferable to remove insoluble matter from this hydrolyzate and further remove water to form it into a solid form. Any method may be used to remove insoluble matter and water as long as the obtained hydrolyzate does not undergo denaturation or thermal decomposition, such as filtration, centrifugation, centrifugal filtration, spray drying, Any method such as spray cooling, drum drying, vacuum drying, freeze drying, etc. can be used. Alternatively, it can also be obtained as a solid by binding or supporting a water-soluble component on a suitable carrier and then removing the solvent by the method described above.

The wheat protein hydrolyzate used in the present invention is commercially available (for example, "Glutamine Peptide GP-1", "Glutamine Peptide GP-2", "Glutamine Peptide GP-1N" manufactured by Nisshin Pharma Co., Ltd.) etc.) may be used as is, or may be used as a raw material after further treatment such as hydrolysis and fractionation.

A preferred embodiment of the wheat protein hydrolyzate used in the present invention is one containing 65 to 90% by mass of glutamine-containing peptides, and the proportion of glutamine and glutamic acid in the peptide is 30 to 60% by mass.

The amount of glutamine-containing peptides in wheat protein hydrolyzate is 18 types (Arg, Lys, His, Phe, Tyr, Leu, Ile, Met, Val, Ala, Gly, Pro, Glu+Gln, Ser, Thr , Asp+Asn, Trp, Cys). By having glutamine-containing peptides in the wheat protein hydrolyzate of 65% by mass or more, it is possible to have a relatively high content of peptide components, which has the advantage of being effective with a small amount of intake in order to obtain the TGF-β suppressing effect. be. The proportion of glutamine-containing peptides in the wheat protein hydrolyzate should be 90% by mass or less to ensure stability, such as suppressing browning due to moisture absorption, and ease of administration (high content of glutamine-containing peptides improves taste). This is preferable in terms of (suppression of deterioration of) Therefore, the content of glutamine-containing peptides in the wheat protein hydrolyzate is preferably 65 to 90% by weight, more preferably 70 to 88% by weight, particularly preferably 70 to 85% by weight. The proportion of glutamine-containing peptides in the wheat protein hydrolyzate can be measured by the method described in the Examples below.
In addition, in the wheat protein hydrolyzate, the remainder other than the glutamine-containing peptide includes carbohydrates, ash, free amino acids, and the like.

The proportion of glutamine and glutamic acid in the glutamine-containing peptide refers to the proportion of Glu+Gln in the total amount of the 18 types of amino acids mentioned above. When the proportion of glutamine and glutamic acid in the glutamine-containing peptide is 30% by mass or more, the proportion of glutamine and glutamic acid is relatively high, and there is an advantage that the effects of glutamine and glutamic acid can be easily obtained. If the proportion of glutamine and glutamic acid in the peptide is 60% by mass or less, there is an advantage that the types of amino acids constituting the peptide component are relatively increased, and the effects of the peptide component can be easily obtained. Furthermore, since increasing the content of glutamine and glutamic acid leads to a decrease in production yield, there is an advantage that production becomes easier. Therefore, the proportion of glutamine and glutamic acid in the peptide is preferably 30 to 60% by weight, more preferably 32 to 55% by weight, particularly preferably 35 to 50% by weight. The total proportion of glutamine and glutamic acid in the glutamine-containing peptide can be measured by the method described in the Examples below.

The wheat protein hydrolyzate preferably contains 30 to 70% by mass of peptides having a molecular weight of 5,000 or more. The ratio of peptides with a molecular weight of 5000 or more in the wheat protein hydrolyzate is 30% by mass or more, which has the advantage that the effect as a peptide component is more likely to be obtained, rather than as an aggregate of individual amino acids. The ratio of peptides with a molecular weight of 5,000 or more in the wheat protein hydrolyzate is 70% by mass or less, which has the advantage that the effect as a peptide component is easily obtained rather than as an effect as a protein. Therefore, the proportion of peptides having a molecular weight of 5000 or more in the wheat protein hydrolyzate is preferably 30 to 70% by mass, more preferably 30 to 65% by mass, particularly preferably 30 to 60% by mass. The proportion of peptides with a molecular weight of 5000 or more in the wheat protein hydrolyzate can be measured by the method described in the Examples below. Note that the proportion of peptides with a molecular weight of 5,000 or more in the wheat protein hydrolyzate measured by the method described in the Examples below can be regarded as the proportion of peptides with a molecular weight of 5,000 or more in the glutamine-containing peptide.

The average molecular weight of the wheat protein hydrolyzate is preferably 200 to 100,000 from the viewpoint of ease of production, and more preferably 500 to 20,000. The average molecular weight can be measured by the method described in Examples below.

The wheat protein hydrolyzate used in the present invention is derived from wheat that has been eaten and is composed of L-amino acids that constitute natural proteins. It is also preferable from the viewpoint of ease of preparation, economical efficiency, etc. Furthermore, the wheat protein hydrolyzate used in the present invention has good storage stability and excellent solubility, so it can be ingested in various forms such as food, feed, and medicine.

The composition for inhibiting TGF-β of the present invention is administered in the range of 0.5 to 15 g, preferably 1 to 10 g per day for adults, based on the dry weight of the wheat protein hydrolyzate. In particular, in the present invention, from the viewpoint of enhancing the TGF-β suppressing effect, the dry mass of the wheat protein hydrolyzate is preferably 3.5 g or more, more preferably 4.5 g or more per day for an adult, It is particularly preferable that the amount is 6.0 g or more.

The intake amount of the glutamine-containing peptide of the composition for suppressing TGF-β of the present invention is preferably 0.33 to 13.5 g, more preferably 0.7 to 9 g per adult. Particularly in the present invention, from the viewpoint of enhancing the TGF-β suppressing effect, the amount of glutamine-containing peptide per adult per day is preferably 2.3 g or more, more preferably 4.0 g or more, and 4.2 g or more. It is particularly preferable.

The composition for suppressing TGF-β of the present invention may be taken in the above amount continuously for several days, for example, 2 to 10 days, or temporarily, for example, for only one day.

The composition for suppressing TGF-β of the present invention is made from safe wheat and does not undergo excessive refining operations, so it is highly safe, can be taken orally as is, and can be used for a long period of time. Continuous intake is possible. Furthermore, since it has good storage stability and excellent solubility, it can be easily formed into compositions for various foods, feeds, medicines, and the like.

At that time, the wheat protein hydrolyzate obtained in the above step can be used alone as it is, but other components exhibiting TGF-β suppressing activity may also be used in combination, as long as they do not impair the effects of the present invention. can. Other components that exhibit TGF-β suppressive effects include, for example, theanine; lactic acid bacteria (Lactococcus lactis subsp. cremoris H-61); anti-TGF-β antibodies; leucine, phenylalanine, α-aminoisobutyric acid, N-methylalanine, - Peptides consisting of methyl isoleucine; imide/amide ether compounds, and phenylacetate and its derivatives. When these other components exhibiting a TGF-β suppressing effect are used together, the total amount is preferably 100 parts by mass or less, more preferably 75 parts by mass or less, per 100 parts by mass of the wheat protein hydrolyzate. The amount is preferably 50 parts by mass or less, particularly preferably 50 parts by mass or less.

The present inventors have found that when the composition for suppressing TGF-β of the present invention contains potassium citrate, the taste is significantly improved and it becomes easier to take orally. For example, in the present invention, in order to enhance the TGF-β suppressing effect, it is preferable to use a relatively large amount (for example, 3.5 g or more per adult per day) of wheat protein hydrolyzate, but if it contains potassium citrate, When blended with wheat protein hydrolyzate, the taste is improved and the above amount becomes easier to drink. Monopotassium citrate and tripotassium citrate are known for oral use as potassium citrate, but it is preferable to use tripotassium citrate in the present invention.

When the composition for suppressing TGF-β of the present invention contains potassium citrate, the amount of potassium citrate is 2 parts by mass or more based on 100 parts by mass of the dry weight of the wheat protein hydrolyzate. The amount is preferably 20 parts by mass or less in terms of improving the taste, and is preferably 20 parts by mass or less. From this point of view, the amount of potassium citrate is preferably 2 to 20 parts by weight, more preferably 5 to 17 parts by weight, and 8 to 14 parts by weight per 100 parts by weight of the dry wheat protein hydrolyzate. Parts by mass are particularly preferred.

Further, in the present invention, when potassium citrate is contained, it is more preferable to combine it with sodium citrate in terms of taste. Although disodium citrate and trisodium citrate are known for oral use as sodium citrate, it is preferable to use trisodium citrate in the present invention. When the composition for suppressing TGF-β of the present invention contains sodium citrate in addition to potassium citrate, the amount of sodium citrate is 75 parts by mass or more based on 100 parts by mass of potassium citrate. The amount is preferably 250 parts by mass or less from the viewpoint of improving the taste, and is preferably 250 parts by mass or less from the viewpoint of improving the taste. From this viewpoint, the amount of sodium citrate is preferably 75 to 250 parts by mass, more preferably 100 to 225 parts by mass, and preferably 125 to 200 parts by mass with respect to 100 parts by mass of potassium citrate. Particularly preferred.

In addition, in the present invention, the amount of citric acid, sodium citrate, and potassium citrate refers to the amount excluding water of hydration when these are hydrates.

The composition of the present invention contains at least one selected from citric acid, magnesium, calcium, and potassium, so that when the composition of the present invention is ingested before or during exercise, it has a TGF-β suppressing effect and a heat-stimulating effect. It is preferable in that it can increase resistance to disease and further contribute to prolonging exercise time, and it is particularly preferable to contain citric acid and at least one selected from magnesium, calcium, and potassium, and especially citric acid and magnesium. It is preferable to contain and/or calcium, and it is especially preferable to contain citric acid, magnesium, calcium, and potassium. Citric acid may be in the form of a citrate salt. Examples of citrates include potassium citrate, sodium citrate, iron citrate, and calcium citrate. Magnesium, calcium and potassium may each be in the form of a compound containing the metal. Examples of magnesium compounds include magnesium sulfate, magnesium chloride, magnesium carbonate, magnesium oxide, magnesium hydroxide, magnesium hydrogen phosphate, magnesium gluconate, magnesium silicate, magnesium aluminate metasilicate, magnesium aluminate silicate, etc. Magnesium sulfate, magnesium chloride, and magnesium carbonate are preferred from the viewpoint of taste. Examples of calcium compounds include calcium lactate, calcium phosphate, calcium citrate, calcium chloride, calcium carbonate, calcium hydroxide, calcium hydrogen phosphate, calcium gluconate, and calcium silicate. Calcium acid and calcium chloride are preferred. Examples of the potassium compound include potassium chloride, potassium nitrate, potassium citrate, potassium gluconate, and acesulfame potassium. Potassium citrate, acesulfame potassium, and potassium chloride are preferred from the viewpoint of taste. The amount of citric acid (in the case of citrate, citric acid ion The standard amount) is preferably 10 parts by mass or more and 100 parts by mass or less, more preferably 15 parts by mass or more and 75 parts by mass or less. From the same point of view, the amount of magnesium (based on metal) is preferably 0.01 parts by mass or more and 20 parts by mass or less, and 0.1 parts by mass or more and 10 parts by mass based on 100 parts by mass of the dry mass of the wheat protein hydrolyzate. Part or less is more preferable. From the same viewpoint, the amount of calcium (based on metals) is preferably 0.01 parts by mass or more and 20 parts by mass or less, and 0.1 parts by mass or more and 10 parts by mass relative to 100 parts by mass of the dry mass of the wheat protein hydrolyzate. The following are more preferable. From the same point of view, the amount of potassium (based on metals) is preferably 0.01 parts by mass or more and 20 parts by mass or less, and 0.1 parts by mass or more and 10 parts by mass relative to 100 parts by mass of the dry mass of the wheat protein hydrolyzate. The following are more preferable.

The composition of the present invention can contain saccharides. Saccharide is a general term for polyhydric alcohols having one carbonyl group (aldehyde group or ketone group). Saccharides include monosaccharides, disaccharides, oligosaccharides, sugar alcohols, and polysaccharides. Examples of monosaccharides include hexoses such as glucose, galactose, mannose, and fructose, and pentoses such as arabinose; examples of disaccharides include maltose, trehalose, lactose, and sucrose; and examples of oligosaccharides include fructo-oligos. Examples of sugar alcohols include maltitol, erythritol, mannitol, sorbitol, xylitol, and the like; examples of polysaccharides include starch, amylose, amylopectin, pullulan, and dextrin. Disaccharides or sugar alcohols are preferred in terms of taste, and at least one selected from maltose, trehalose, maltitol, mannitol, and sorbitol is more preferred. When the composition of the present invention contains saccharides, the amount thereof is not particularly limited, but from the viewpoint of taste, for example, 1 part by mass or more and 100 parts by mass per 100 parts by mass of the dry wheat protein hydrolyzate. The amount is preferably 5 parts or more and 50 parts by mass or less, more preferably 5 parts by mass or more and 50 parts by mass or less.

The content of wheat protein hydrolyzate in the composition of the present invention varies depending on its dosage form, but whether the composition of the present invention is a food product or a non-food product, such as a pharmaceutical product, the content of the wheat protein hydrolyzate varies depending on the dosage form. It is preferably in the range of 10 to 75% by weight, more preferably 15 to 70% by weight, particularly preferably 20 to 65% by weight, based on dry weight. It is desirable to have a form in which the daily dosage can be controlled so that the above-mentioned daily intake for adults can be taken.

The composition for suppressing TGF-β of the present invention can be used as a food for suppressing TGF-β. Foods for suppressing TGF-β include the above-mentioned wheat protein hydrolysates, sodium citrate, potassium citrate, other citric acid-containing compounds, magnesium compounds, calcium compounds, potassium compounds, and sugars. In addition, other food materials, various nutrients, various vitamins, minerals, amino acids, various oils and fats, various additives (e.g., flavor components, sweeteners, acidulants such as organic acids, surfactants, pH adjusters, It can be manufactured according to a conventional method by adding stabilizers, antioxidants, pigments, flavors), etc. Foods for suppressing TGF-β include health foods, foods with functional claims, foods for specified health uses, and all foods that can contain wheat protein hydrolysates. The composition for suppressing TGF-β and the food for suppressing TGF-β of the present invention can be in various formulations such as tablets, chewable tablets, powders, capsules, granules, drinks, and liquid foods such as enteral nutrition. It can be done. Foods in the form of formulations can be produced in the same manner as pharmaceuticals described below.

For example, the TGF-β suppressing food according to the present invention can be produced by blending a wheat protein hydrolyzate into a commonly eaten food. For example, the food for suppressing TGF-β according to the present invention includes beverages. Furthermore, the TGF-β suppressing food according to the present invention includes tea drinks such as green tea, oolong tea, and black tea, soft drinks, jelly drinks, sports drinks, milk drinks, carbonated drinks, fruit juice drinks, lactic acid bacteria drinks, fermented milk drinks, and powder drinks. , cocoa drinks, beverages such as purified water, spreads such as butter, jam, furikake, margarine, mayonnaise, shortening, custard cream, dressings, breads, rice, noodles, pasta, miso soup, tofu, milk, yogurt, It may be prepared as soups or sauces, confectionery (eg, biscuits, cookies, chocolate, candy, cake, ice cream, chewing gum, tablets), etc.

The TGF-β suppressing food according to the present invention includes not only food for humans but also feed for livestock, racehorses, etc., pet food, and the like. Since feed is almost the same as food except that the target is an animal, the description herein regarding foods for suppressing TGF-β can be similarly applied to feed.

When the composition of the present invention is a pharmaceutical, its dosage form includes oral preparations such as tablets, capsules, granules, powders, syrups, dry syrups, solutions, suspensions, inhalants, suppositories, etc. Examples include enteral preparations, ointments, creams, gels, skin external preparations such as patches, drops, and injections. Among these, oral preparations are preferred.

When the composition according to the present invention is a pharmaceutical, commonly used additives such as excipients, disintegrants, binders, lubricants, surfactants, Alcohol, water, a water-soluble polymer, a sweetener, a flavoring agent, an acidulant, etc. can be mixed according to the dosage form, and the formulation can be prepared according to a conventional method. Liquid preparations such as solutions and suspensions may be dissolved or suspended in water or other suitable medium immediately before administration, and in the case of tablets and granules, they may be dissolved or suspended in water or other suitable media immediately before administration. The surface may be coated.

As shown in the Examples described below, the TGF-β suppressing composition and TGF-β suppressing food of the present invention can reduce the TGF-β concentration in blood, especially plasma, by ingesting the same. . Due to this action, specific effects based on TGF-β suppressive activity include chronic glomerulonephritis, renal interstitial fibrosis, diabetic nephropathy, liver fibrosis, liver cirrhosis, pulmonary fibrosis, keloids, and scleroderma. , arteriosclerosis, posterior myocardial infarction, cardiac fibrosis, restenosis after angioplasty, acute megakaryoblastic leukemia, adult T-cell leukemia, etc. can be expected to have preventive or therapeutic effects. Furthermore, by ingesting the composition for suppressing TGF-β and the food for suppressing TGF-β of the present invention, it is possible to reduce the subjective intensity of exercise load felt during exercise load. Therefore, by ingesting the composition for suppressing TGF-β and the food for suppressing TGF-β of the present invention, one can exercise while feeling less burdened, and can improve exercise habits such as extending exercise time. The composition for suppressing TGF-β and the food for suppressing TGF-β of the present invention may be taken before exercise, during exercise, or after exercise, but it is more effective to take it before exercise, It is more preferable to take it orally the day before exercise, or the day before and several days before exercise.
In addition, as shown in the examples below, the composition for suppressing TGF-β and the food for suppressing TGF-β of the present invention have remarkable taste properties, especially when containing potassium citrate, and have excellent taste properties and TGF-β suppression properties. It has both β-suppressing ability and safety, and is also effective in preventing heatstroke.

EXAMPLES Hereinafter, the present invention will be explained in more detail with reference to examples, but the present invention is not limited to the following examples.

(Example 1)
28.00 kg of wheat gluten hydrolyzate (“GP-1N” manufactured by Nisshin Pharma), 6.85 kg of trehalose (manufactured by Hayashibara), 10.50 kg of citric acid anhydride (manufactured by Showa Kako), trisodium citrate dihydrate ( (manufactured by Showa Kako) 6.24 kg, tripotassium citrate (manufactured by Showa Kako) 3.50 kg, magnesium sulfate heptahydrate (manufactured by Tomita Pharmaceutical) 1.17 kg, calcium lactate pentahydrate (manufactured by Taihei Kagaku Sangyo) 2. 33 kg were mixed and granulated using a fluidized bed granulator (manufactured by Powrex) to obtain 69.59 kg of granulated powder. The binder for granulation is made by dissolving 7.00 kg of wheat gluten hydrolyzate ("GP-1N" manufactured by Nisshin Pharma), 0.99 kg of pullulan (manufactured by Hayashibara), and 0.70 kg of acesulfame potassium (manufactured by MC Food) in purified water. The entire amount was sprayed. 1.51 kg of antifoaming agent and 0.80 kg of fragrance were mixed into the granulated powder.
Through the above steps, a composition containing 6 g of wheat gluten hydrolyzate in a daily dose of 12 g was obtained.
As a result of analysis using the method described below, the proportion of glutamine-containing peptides in the wheat gluten hydrolyzate was 70% by mass (4.2 g per 12 g daily dose), and among the constituent amino acids of the glutamine-containing peptides, glutamine and glutamic acid were The proportion was 40% by mass. It was also confirmed that the wheat gluten hydrolyzate contained 32% by mass of peptides with a molecular weight of 5,000 or more, and that the average molecular weight was 4,700. In the daily dose of 12 g of the composition obtained in Example 1, potassium citrate was 0.6 g and sodium citrate was 0.94 g. Furthermore, in the daily dose of 12 g of the composition obtained in Example 1, the amount of citric acid was 2.7 g, magnesium 28.0 mg (metal basis amount), calcium 56.0 mg (metal basis amount), potassium ( It was confirmed that the amount (based on metal) was 260 mg.

Note that the amount of citric acid was measured by high performance liquid chromatography. For sample preparation, an appropriate amount of sample was taken, 5% (v/v) perchloric acid and water were added, and extraction was performed by homogenization or shaking. Further water was added to make the volume constant, filtered, and analyzed using an HPLC system.

The amounts of magnesium and calcium were measured by ICP emission spectrometry. The sample was prepared by dry ashing, extraction with hydrochloric acid, filtration, and then adjusting the volume appropriately.

The amount of potassium was measured by atomic absorption spectrophotometry. Samples were prepared by a hydrochloric acid extraction method in which diluted hydrochloric acid was added, extracted by shaking, and filtered, or by a dry ashing method.

(Amount of glutamine-containing peptide in wheat protein hydrolyzate)
(Amount of glutamine and glutamic acid in glutamine-containing peptide)
Glutamine-containing peptides in wheat gluten hydrolyzate are amino acids (18 types; Arg, Lys, His, Phe, Tyr, Leu, Ile, Met, Val, Ala, Gly, Pro, Glu+Gln, Ser, Thr, Asp). +Asn, Trp, Cys) was quantified as the total amount. Specifically, it was measured as follows.

Samples prepared by the following method were analyzed using an automatic amino acid analyzer (JLC-500/V or JLC-500/V2 manufactured by JEOL Ltd.) and a high-performance liquid chromatograph (LC-20AD manufactured by Shimadzu Corporation) to analyze 18 types (Arg, Lys, , His, Phe, Tyr, Leu, Ile, Met, Val, Ala, Gly, Pro, Glu+Gln, Ser, Thr, Asp+Asn, Trp, Cys). The columns used were LCR-6 (diameter 4 mm x length 120 mm) manufactured by JEOL Ltd. and CAPCELL PAK C18 AQ (diameter 4.6 mm x length 250 mm) manufactured by Osaka Soda. As the mobile phase, "sodium citrate buffer (H-01 to H-04)" or "lithium citrate buffer (P-21)" manufactured by JEOL Ltd., a mixture of 20 mmol/L perchloric acid and methanol. (volume ratio 80:20) was used. As a reaction solution for automatic amino acid analysis, "Ninhydrin Coloring Solution Kit-II for JEOL" manufactured by Fuji Film Wako Pure Chemical Industries, Ltd. was used.
Samples were prepared using the following method.
0.2 to 1.5 g of the sample was accurately weighed into a sealed test tube, and 20 mL of 6 mol/L hydrochloric acid (containing 0.04% (v/v) mercaptoethanol) was added. After degassing for 15 minutes under reduced pressure of 2.0 kPa or less, the tube was sealed and hydrolysis was performed at 110° C. for 24 hours. After the hydrolysis, the solution was cooled and opened, and the hydrolyzed solution was transferred to a 100 mL volumetric flask and made up to volume with water to adjust the concentration. The fixed volume of the hydrolyzed solution was adjusted to pH 2.2 with 3 mol/L sodium hydroxide solution, the fixed volume was made up with 0.067 mol/L sodium citrate buffer (pH 2.2), and the solution was filtered through a 0.45 μm filter.
The total amount of the 18 types of amino acids obtained above was taken as the ratio of the glutamine-containing peptide. Furthermore, the proportion of glutamine and glutamic acid in the glutamine-containing peptide was calculated from the amount of Glu+Gln obtained above.
The amount of wheat gluten hydrolyzate used as the denominator was the mass measured using a precision balance.

(Percentage of peptides with a molecular weight of 5000 or more in wheat protein hydrolyzate)
It was determined by measuring the molecular weight distribution using HPLC method.
An HPLC system using the gel filtration column for peptides described below was set up, a known peptide serving as a molecular weight marker was charged, and a calibration curve was determined based on the relationship between molecular weight and retention time. To prepare the sample, take approximately 0.02 g of the sample, add 10 mL of a mixture of water, acetonitrile, and trifluoroacetic acid (volume ratio 55:45:0.1), leave it at room temperature overnight, and pass it through a 0.45 μm filter. Filtered. 20 μL of the sample solution was applied to HPLC. The proportion (%) of the peptide fraction with a molecular weight of 5,000 or more in the wheat protein hydrolyzate was determined by the ratio of the area in the range of molecular weights of 5,000 or more (time range) to the area of the entire absorbance chart. Further, the weight average molecular weight in the molecular weight range of 200 to 100,000 was defined as the average molecular weight.
As the column, TSKgel G2500PW _XL (diameter 7.8 mm x length 300 mm) manufactured by Tosoh Corporation was used.
As the mobile phase, a mixed solution of water, acetonitrile and trifluoroacetic acid (volume ratio 55:45:0.1) was used.
The column temperature was 40° C., the flow rate was 0.5 ml/min, and the detection wavelength was 220 nm.
Cytochrome C (molecular weight 12327) using the following as a molecular weight marker
Aprotinin (molecular weight 6512)
Bacitracin (molecular weight 1450)
Angiotensin II (molecular weight 1046)
Gly-Gly-Tyr-Arg (molecular weight 451)
Gly-Gly-Gly (molecular weight 189)

(Comparative example 1)
The control composition was a placebo containing an excipient (dextrin) instead of the wheat gluten hydrolyzate, and a composition that had the same raw materials and manufacturing method as the composition manufactured in Example 1 except for the wheat gluten hydrolyzate. (Comparative Example 1). It was confirmed that the two compositions were indistinguishable in appearance, flavor, and properties, and were used in the test in a blinded manner.

(Plasma TGF-β concentration)
A placebo-controlled, randomized, double-blind, crossover comparative study was conducted on 18 healthy Japanese men and women (14 men, 4 women) aged 20 to under 65. The 18 subjects were assigned using a stratified randomization method and divided into two groups (Group A: 9 subjects, Group B: 9 subjects).
Group A was orally ingested 12 g of the composition of Example 1 dissolved in 500 mL of water every day for 8 days, followed by a 2-week rest period, and then ingested the composition of Comparative Example 1 for 8 days. 12 g per day was dissolved in 500 mL of water and administered orally.
Group B was orally ingested 12 g of the composition of Comparative Example 1 dissolved in 500 mL of water every day for 8 days, followed by a 2-week rest period, and then administered the composition of Example 1 for 8 days. 12 g per day was dissolved in 500 mL of water and administered orally.
In both groups A and B, during the test period of 8 days + 2 weeks + 8 days, the subjects were asked not to exercise for more than 30 minutes more than twice a week.
On the start day of the 8-day intake period (before intake) and on the end day of the intake period (after intake), exercise was performed on a bicycle ergometer (Aerobic Bike (registered trademark) 75XLIII (Konami Sports Life Co., Ltd.)) for 60 minutes. Immediately after the end of the exercise load, the concentration of total TGF-β in plasma was measured by the following method. Plasma TGF-β concentration was measured by ELISA method. The results are shown in Table 1.

Ingestion of the composition of Example 1 significantly decreased the TGF-β concentration in plasma compared to before ingestion. In a comparison between groups, after ingestion, the group ingesting the composition of Example 1 showed a significantly lower plasma TGF-β concentration compared to the group ingesting the composition of Comparative Example 1 (P= 0.027).

(Subjective exercise intensity)
In the above-mentioned (plasma TGF-β concentration) test, 14 men (6 men in the first group A, 8 men in the B group) were subjected to subjective tests immediately before and after the exercise load before and after the above intake. The participants were asked to evaluate their exercise intensity. Subjective exercise intensity was graded on a scale of 15 from 6 to 20 (6: at rest, 7: very easy, 9: quite easy, 11: easy, 13: somewhat hard, 15: The subjects were asked to indicate which grade their condition during exercise corresponded to (hard, 17: quite hard, 19: very hard), and the results were recorded. Table 2 shows the average scores of the composition intake group of Example 1 and the composition intake group of Comparative Example 1.

Ingestion of the composition of Example 1 significantly lowered the subjective exercise intensity score immediately after exercise load compared to before ingestion (P=0.0203). In a comparison between the groups, after ingestion, the group ingesting the composition of Example 1 showed a significantly lower score immediately after exercise load compared to the group ingesting the composition of Comparative Example 1 (P=0 .0481).

(Example 2)
In Example 1, the entire amount of potassium citrate was changed to anhydrous citric acid. A composition of Example 2 was produced in the same manner as Example 1 except for this point.

(Example 3)
In Example 1, the entire amount of potassium citrate was changed to sodium citrate. The composition of Example 3 was produced in the same manner as Example 1 except for this point.

(Example 4)
In Example 1, the entire amount of potassium citrate was changed to potassium chloride. The composition of Example 4 was produced in the same manner as Example 1 except for this point.

(Example 5)
In Example 1, potassium citrate was changed to trehalose.
A composition of Example 5 was produced in the same manner as Example 1 except for this point.

(taste)
Taste was evaluated by sensory evaluation by a panel of 5 healthy Japanese people (3 men, 2 women) aged 20 to 65 years. The sensory evaluation was scored based on the following criteria, and the average value of the five participants was calculated. The results are shown in Table 3. The amount of each sample ingested by one panelist was as follows: 12 g of the compositions of Examples 1 to 5 were dissolved in 500 mL of water, and 50 to 200 mL was orally ingested.
5: Very good balance of taste, sourness, and sweetness derived from wheat protein hydrolyzate.
4: Good balance of taste derived from wheat protein hydrolyzate, sourness, and sweetness.
3: The taste derived from wheat protein hydrolyzate, sourness, and sweetness have a normal balance.
2: The taste derived from wheat protein hydrolyzate, sourness, and sweetness are unbalanced.
1: The taste derived from wheat protein hydrolyzate, sourness, and sweetness are very unbalanced.

Claims (2)

A composition for suppressing TGF-β containing a wheat protein hydrolyzate , the wheat protein hydrolyzate containing 65 to 90% by mass of a glutamine-containing peptide, and the proportion of glutamine and glutamic acid in the peptide being 30% by mass. Using a hydrolyzate of wheat gluten with protease and amylase, which contains ~60% by mass and 30 to 70% by mass of peptides with a molecular weight of 5000 or more,
Contains sodium citrate, potassium citrate, anhydrous citric acid, magnesium sulfate, sugars and sweeteners,
A composition for suppressing TGF-β, which is used to suppress an increase in TGF-β in plasma during exercise . A food for suppressing TGF-β, comprising the composition according to claim 1.