TW201842924A - Composition for promoting increase in BDNF level in body - Google Patents

Abstract

The present invention provides a composition for promoting an increase in the level of brain-derived neurotrophic factor (BDNF) in the body, especially in the muscles. This composition for promoting an increase in the level of BDNF in the body has as an active ingredient a whey protein hydrolysate, wherein the composition is characterized in that the whey protein hydrolysate contains at least one dipeptide selected from the group consisting of Ile-Leu, Leu-Leu, and Val-Leu.

Description

a composition for promoting an increase in the amount of BDNF in the body

The present invention relates to a composition having an effect of promoting an increase in the amount of Brain Derived Neurotrophic Factor (BDNF) (hereinafter, simply referred to as "BDNF" in the present invention). In more detail, the present invention relates to a composition which promotes an increase in the amount of BDNF in the body, particularly in muscle, with or without exercise.

Now, in Japan, the increase in mental illness such as depression and dementia (including Alzheimer's type dementia and vascular dementia) has become a social problem. The number of elderly dementia patients has doubled in 10 years since the 1.49 million in 2002, and reached more than 3 million in 2012. In addition, according to the report of the Research Office of the Ministry of Health, Labour and Welfare of Japan (2012), it is estimated that about 4 million people with mild dementia who are at risk of being affected by the onset of dementia are calculated to be over 65 years old and 4 people. One person will become a person with dementia or dementia. The current situation is that once the dementia is ill, even if it is treated, it can only inhibit the progression of the disease and cannot cure it. Therefore, once the dementia is ill, not only will the patient's life change drastically, but also the family member will be overburdened with care and will have to be forced to leave because of care. The change in life plan cannot be avoided. The impact of the labor loss of the labor generation caused by the departure of the company on the national economy cannot be ignored.

Therefore, prevention and control alone does not make the onset of dementia not only a critical issue for individuals, but also a critical issue for modern society.

It is traditionally believed that the intake of milk and dairy products is useful for the prevention of this dementia. Specifically, there are reports or inferences that calcium and magnesium rich in milk and dairy products have a preventive effect on dementia (Non-Patent Document 1), and whey contained in milk and dairy products is improved as Azhai. Insulin resistance of a silent dementia risk factor (Non-Patent Document 2).

On the other hand, it is known that once exercised, BDNF, which is one of the neurotrophic factors involved in the production, growth, maintenance, or regeneration of nerve cells, is significantly increased, and the performances of memory, learning, and the like are simultaneously improved (Non-Patent Document 3) ). Further, according to many epidemiological studies and experiments, it is known that the cognitive function of the elderly can be suppressed by the movement (Non-Patent Documents 4 to 6). Further, it has recently been confirmed by animal experiments that hippocampal BDNF is increased by forced exercise load through temporary electrical stimulation (Non-Patent Document 7). In addition, it has been reported that the amount of expression of BDNF in the brain is reduced in patients with depression and Alzheimer's type dementia (Non-Patent Document 8 and Non-Patent Document 9).

Therefore, it is suggested that BDNF which is increased by exercise is effective for preventing mental illness such as depression and dementia.

In addition, it has been reported that by administering BDNF intracerebroventricularly, subcutaneously or intraperitoneally, the increase in body weight is suppressed and the glucose metabolism is improved (Non-Patent Document 10). [Prior Technical Literature] [Non-Patent Literature]

[Non-Patent Document 1] J Am Geriatr Soc. 2012, 60(8): 1515-1520. [Non-Patent Document 2] Nutrition. 2014, 30(9): 1076-1080 [Non-Patent Document 3] Neeper SA et al Brain Res, 726: 49-56 (1996) [Non-Patent Document 4] Trends Neurosci. 2002, 25(6): 295-301. [Non-Patent Document 5] Nature. 1999, 29; 400 (6743): 418 -419. [Non-Patent Document 6] Cell Mol Neurobiol. 2010, 30(4): 493-503. [Non-Patent Document 7] Japan 31st Subsidized Young Researcher Health Science Research Report 2014, pp.104- 109 [Non-Patent Document 8] Chen B et al, Biolpsychiatry. 50: 260-265 (2001). [Non-Patent Document 9] Phillips HS et al. Neuron. 7: 695-702 (1991). [Non-Patent Document 10 ] Nakagawa T et al, Diabetes, 49: 436-444 (2000)

[Problems to be solved by the invention]

As mentioned above, studies on exercise and cognitive function have flourished, and there are results suggesting that BDNF is associated with improved cognitive function brought about by exercise. However, the effects of dietary intake on BDNF, especially the combination of exercise and intake of food and drink. There are still many unclear parts of the effect. Therefore, the present invention is directed to the effect of eating food and drink on BDNF, especially the effects of exercise and intake of food and drink. Specifically, the present invention has been developed in order to efficiently increase the amount of BDNF involved in the improvement of cognitive function and glucose metabolism by ingestion of food or drink or in combination with exercise and food and drink.

That is, the present invention aims to provide a composition for promoting an increase in the amount of BDNF in the body, particularly in the muscle, in other words, to provide a composition for promoting an increase in the amount of BDNF in the body, particularly in the muscle. Furthermore, the present invention aims to provide a method for promoting an increase in the amount of BDNF in the body, particularly in the muscle. [Means to solve the problem]

In order to solve the above problems, the inventors of the present invention found that the amount of BDNF in the muscle is obtained by ingesting a whey protein hydrolyzate containing at least one dipeptide selected from the group consisting of Ile-Leu, Leu-Leu, and Val-Leu. Will increase significantly. Since this effect was not observed clearly in the whey protein before hydrolysis, it is presumed that this effect comes from the contribution of the dipeptide which is not contained in the whey protein as an intrinsic component of the whey protein hydrolyzate. As mentioned above, there are reports suggesting that BDNF is effective for the prevention of mental illness such as depression or dementia as well as for cognitive improvement, and that the inhibition of weight gain and the improvement of glucose metabolism are also effective. Therefore, it is presumed that the dipeptide-containing whey protein hydrolyzate is useful for prevention of mental illness such as depression or dementia, improvement of cognitive function, suppression of weight gain, and/or improvement of glucose metabolism.

The present invention has been completed based on these findings and has the following embodiments. In the following description, the whey protein hydrolysate is simply referred to as "WPH". Further, the dipeptide is a three-letter symbol using an amino acid, and the amino acid sequence constituting the dipeptide is described as "Xxx-Yyy".

(I) A composition for promoting an increase in the amount of BDNF in the body (I-1) A composition for promoting an increase in the amount of BDNF, which is an active ingredient for promoting the body, and an increase in the amount of BDNF which is preferably intramuscular. a composition, and the WPH is selected from the group consisting of Ile-Leu (isoleucine thiol-leucine), Leu-Leu (leucine thiol-leucine), and Val-Leu (amidamine-white) At least one dipeptide group in the group of amino acids. (I-2) A composition for promoting an increase in the amount of BDNF according to (I-1), wherein the WPH is those containing Ile-Leu, Leu-Leu, and Val-Leu. (I-3) A composition for promoting an increase in the amount of BDNF according to (I-1) or (I-2), wherein the WPH is further contained from the group consisting of Ile-Val (iso-amine thiol-proline) At least one group of Ile-Ile (iso-amine decyl-isoleucine), Leu-Val (leucine decyl-proline), and Leu-Ile (alkamine-isoleucine) A dipeptide person. (I-4) A composition for promoting an increase in the amount of BDNF as in (I-1) or (I-2), wherein the above WPH is further contained by Ile-Val, Ile-Ile, Leu-Val, and Leu-Ile The dipeptide formed. (I-5) The composition for promoting an increase in the amount of BDNF according to any one of (I-1) to (I-4), which is administered orally to a mammal containing humans before or during exercise or Make it ingest. (I-6) A composition for promoting an increase in the amount of BDNF as in (I-5), which is a composition for promoting an increase in the amount of BDNF in muscle during exercise or after exercise. (I-7) A composition for promoting an increase in the amount of BDNF according to any one of (I-1) to (I-4), which is administered orally to a mammal comprising a human in a non-moving state. Ingestor. (I-8) The composition for promoting an increase in the amount of BDNF according to any one of (I-1) to (I-7), which is orally administered or caused to a person with cognitive deterioration or a diseased person thereof Ingestor. (I-9) A composition for promoting an increase in the amount of BDNF as in (I-8), which is a composition for inhibiting or improving cognitive function deterioration. (I-10) A composition for promoting an increase in the amount of BDNF according to any one of (I-1) to (I-7), which is a person who is obese or/and has a reduced glucose metabolism, or such a possibility The person who is orally administered or ingested by the patient. (I-11) A composition for promoting an increase in the amount of BDNF as in (I-10), which is a composition for inhibiting or ameliorating obesity, or/and a composition for inhibiting or ameliorating a decrease in glucose metabolism. (I-12) The composition for promoting an increase in the amount of BDNF according to any one of (I-1) to (I-11), which is constituted by a unit packaging type of each administration (ingestion), and The unit package type includes the above-mentioned WPH in a single dose (intake amount) of 0.1 to 40 g in terms of total protein. (I-13) A composition for promoting an increase in the amount of BDNF according to any one of (I-1) to (I-12), which is a food or drink. (I-14) A composition for promoting an increase in the amount of BDNF according to (I-13), which is a health food, a functional food, a nutritional supplement food, a functional food, a specific health food, a food for a patient, Milk powder for infants and young children, pregnant women or breastfeeding women, or food supplements with a statement to promote exercise to increase the amount of BDNF in the body.

(II) Method for increasing the amount of BDNF in the body (II-1) A method for increasing the amount of BDNF in the subject, which is preferably intramuscular, which is administered or ingested into the subject, comprising a substance selected from the group consisting of Ile-Leu a composition of WPH of at least one dipeptide grouped by Leu-Leu and Val-Leu. (II-2) The method according to (II-1), wherein the above WPH is those containing Ile-Leu, Leu-Leu, and Val-Leu. (II-3) The method of (II-1) or (II-2), wherein the WPH further comprises at least one selected from the group consisting of Ile-Val, Ile-Ile, Leu-Val, and Leu-Ile Peptide. (II-4) The method according to (II-1) or (II-2), wherein the WPH is a dipeptide further comprising Ile-Val, Ile-Ile, Leu-Val and Leu-Ile. (II-5) The method according to any one of (II-1) to (II-4) wherein the aforementioned subject is a mammal comprising humans before or during exercise. (II-6) The method of (II-5), which is a method of increasing the amount of BDNF in the muscle during exercise or after exercise. (II-7) The method according to any one of (II-1) to (II-4) wherein the aforementioned subject is a mammal comprising a human in a non-exercise state. (II-8) The method according to any one of (II-1) to (II-7) wherein the aforementioned subject is a cognitive degenerative person or a person thereof. (II-9) The method according to (II-8), which is a method for inhibiting or improving the deterioration of cognitive function of the aforementioned subject. (II-10) The method according to any one of (II-1) to (II-8), which is a method for enhancing the expression of memory or learning of a mammal including a human. (II-11) The method according to any one of (II-1) to (II-7) wherein the aforementioned subject is an obese person or/and a reduced glucose metabolism, or such a likely patient. (II-12) The method according to (II-11), which is a method for inhibiting or improving obesity of the aforementioned subject, or/and inhibiting or improving the reduction of glucose metabolism. (II-13) The method of any one of (II-1) to (II-12), wherein the composition is composed of a unit package type of each administration (intake), and is packaged in the unit The type includes WPH which is 0.1 to 40 g in total protein as a single administration amount (intake amount). (II-14) The method according to any one of (II-1) to (II-13), wherein the composition is a food or drink, a health food, a functional food, a nutritional supplement, a functional food, a specific Health foods, foods for patients, milk powder for infants and young children, milk powder for pregnant women or breastfeeding women, or food supplements with a statement to promote exercise to increase the amount of BDNF in the body.

(III) a composition of WPH (III-1) which is used to promote an increase in the amount of BDNF in the body, which is a composition which is used to promote an increase in the amount of BDNF in the body of the subject, and the composition is WPH having at least one dipeptide selected from the group consisting of Ile-Leu, Leu-Leu, and Val-Leu is used as an active ingredient. (III-2) The composition according to (III-1), wherein the above WPH is those containing Ile-Leu, Leu-Leu, and Val-Leu. (III-3) The composition of (III-1) or (III-2), wherein the WPH further comprises at least one selected from the group consisting of Ile-Val, Ile-Ile, Leu-Val, and Leu-Ile Bipeptide. (III-4) The composition according to (III-1) or (III-2), wherein the WPH is a dipeptide further comprising Ile-Val, Ile-Ile, Leu-Val and Leu-Ile. (III-5) The composition according to any one of (III-1) to (III-4), wherein the aforementioned subject is a mammal comprising humans before or during exercise. (III-6) A composition according to (III-5) which is used to promote an increase in the amount of BDNF in the muscle during exercise or after exercise. (III-7) The composition according to any one of (III-1) to (III-4), wherein the subject is a mammal comprising a human in a non-exercise state. (III-8) The composition according to any one of (III-1) to (III-7), wherein the aforementioned subject is a cognitive degenerative person or a person thereof. (III-9) A composition according to (III-8) which is used to inhibit or ameliorate cognitive function deterioration. (III-10) The composition according to any one of (III-1) to (III-7), wherein the aforementioned subject is an obese person or/and a reduced glucose metabolism, or such a likely patient. (III-11) A composition according to (III-10) which is used to inhibit or ameliorate obesity, or/and to inhibit or ameliorate a decrease in glucose metabolism. (III-12) The composition according to any one of (III-1) to (III-11), which is constituted by a unit packaging type of each administration (ingestion), and is in the unit packaging type The composition containing the above-mentioned WPH in a single dose (intake amount) of 0.1 to 40 g in terms of total protein is contained. (III-13) The composition according to any one of (III-1) to (III-12), which is a food or drink, which is a health food, a functional food, a nutritional supplement food, a functional food, and a specific health care product. Foods, foods for the patient, milk powder for infants and young children, milk powder for pregnant or lactating women, or food supplements with a statement to promote exercise to increase the amount of BDNF in the body.

(IV) Use of WPH for the manufacture of a composition for promoting an increase in the amount of BDNF in the body (IV-1) A use for the purpose of promoting an increase in the amount of BDNF which is preferably intramuscular in the subject The use of WPH of the composition, and the WPH is one containing at least one dipeptide selected from the group consisting of Ile-Leu, Leu-Leu, and Val-Leu. (IV-2) The use of (IV-1) wherein the above WPH is those containing Ile-Leu, Leu-Leu and Val-Leu. (IV-3) The use of (IV-1) or (IV-2), wherein the WPH further comprises at least one selected from the group consisting of Ile-Val, Ile-Ile, Leu-Val, and Leu-Ile Peptide. (IV-4) The use of (IV-1) or (IV-2), wherein the WPH is further a dipeptide consisting of Ile-Val, Ile-Ile, Leu-Val and Leu-Ile. (IV-5) The use according to any one of (IV-1) to (IV-4), wherein the aforementioned subject is a mammal comprising humans before or during exercise. (IV-6) The use of (IV-5), wherein the composition is for promoting an increase in the amount of BDNF in the muscle during or after exercise of the subject. (IV-7) The use of any one of (IV-1) to (IV-4), wherein the subject is a mammal comprising a human in a non-exercise state. (IV-8) The use according to any one of (IV-1) to (IV-7), wherein the aforementioned subject is a cognitive degenerative person or a person thereof. (IV-9) The use of (IV-8), wherein the aforementioned composition is for suppressing or improving cognitive function deterioration. (IV-10) The use according to any one of (IV-1) to (IV-7), wherein the aforementioned subject is an obese person or/and a reduced glucose metabolism, or such a likely patient. (IV-11) The use of (IV-10) for inhibiting or ameliorating obesity, or/and inhibiting or ameliorating a decrease in glucose metabolism. (IV-12) The use of any one of (IV-1) to (IV-11), wherein the composition is composed of a unit package type for each administration (intake), and is packaged in the unit The form includes a composition of the above-mentioned WPH in a single dose (intake amount) of 0.1 to 40 g in terms of total protein. (IV-13) The use of any one of (IV-1) to (IV-12), wherein the composition is a food or drink, a health food, a functional food, a nutritional supplement, a functional food, a specific Health foods, foods for patients, milk powder for infants and young children, milk powder for pregnant women or breastfeeding women, or foods and beverages with a statement to promote exercise to increase the amount of BDNF in the body. [Effects of the Invention]

According to the present invention, there is provided a composition (hereinafter, simply referred to as "the composition of the present invention") for promoting an increase in the amount of BDNF in the body, particularly muscle, using WPH as an active ingredient.

By ingesting the composition of the present invention, or ingesting the composition of the present invention during exercise, it is possible to promote exercise to increase the amount of BDNF in the body, particularly in the muscle, and as a result, the amount of BDNF in the muscle can be effectively increased ( Refer to Test Example 1). As described above, it has been known in the past that exercise can suppress the deterioration of cognitive function in the elderly, and BDNF can improve the performance of memory and learning, and suggests that it is also effective for the prevention of mental illness such as depression and dementia. Therefore, it is possible to further enhance the above effects by ingesting the composition of the present invention during exercise.

In addition, the increase in BDNF not only ends in the case of ingestion of WPH but also increases, and also increases significantly in the absence of exercise (refer to Test Example 2). From this, it is presumed that the increase in BDNF is not related to the presence or absence of exercise, but is a unique effect obtained by ingesting WPH. Further, since such an effect is not remarkable in WPH before hydrolysis, it is presumed that this effect comes from the contribution of the dipeptide which is an intrinsic component of the whey protein hydrolyzate, which is not contained in the whey protein (even in a small amount).

In addition, it has been reported that BDNF has an inhibitory effect on weight gain and an improvement in glucose metabolism. Therefore, according to the present invention, it is expected that the effect can be obtained by ingesting the composition of the present invention.

Further, the WPH contained in the composition of the present invention is a milk protein which has been used as a food material for a long time. Therefore, the composition of the present invention does not have to worry about side effects even if it is continuously administered for a long period of time, and is advantageous in terms of high safety. Further, in the production process of natural cheese, whey protein is usually discharged into waste after industrialization of the raw milk and becomes industrial waste. Since the WPH contained in the composition of the present invention as an active ingredient can reuse the industrial waste whey as a raw material, the present invention can be said to be useful both on the environmental and economical aspects.

The composition (the composition of the present invention) for promoting an increase in the amount of BDNF in the body of the present invention is characterized by comprising at least one double containing a group selected from the group consisting of Ile-Leu, Leu-Leu, and Val-Leu. The WPH of the peptide (hereinafter sometimes referred to as "WPH of the present invention") is used as an active ingredient.

In general, WPH is obtained by hydrolyzing all or part of whey protein as a raw material, and is also called whey peptide.

Whey protein is a mixture of globular proteins contained in whey (whey) which is a liquid fraction after removing casein and fat from milk, and contains α-lactalbumin (α-La) and β-lactoglobulin. Proteins such as (β-Lg), immunoglobulin, lactoferrin, and bovine serum albumin. In the present invention, the whey protein used as a raw material for preparing WPH may be a whey protein purified from milk or whey, or may be a crude whey protein (whey protein containing) separated from milk or whey. ()). Further, a food or drink containing whey protein and a raw material thereof (for example, a composition containing other components other than milk protein, etc.) may be used. The food or drink and the raw material thereof may also be those containing casein in addition to whey protein.

Examples of the purified whey protein include α-lactalbumin (α-La), β-lactoglobulin (β-Lg), immunoglobulin, lactoferrin, and a protein mixture containing at least one of these. There is also whey protein refined product (WPI) and the like. In addition, as the above-mentioned purified whey protein (whey protein content), for example, whey stock solution (sweet whey, acid whey, etc.) prepared from cow's milk, a concentrate thereof, and a dried product (whey powder, etc.) may be mentioned. , its frozen material and such reduced substances. Further, these may include desalted whey, whey protein concentrate (WPC), and the like. In addition, examples of the food or drink containing whey protein or raw materials thereof include raw milk, sterilized milk (milk, etc.), skim milk, component-adjusted milk, processed milk, and dairy products (concentrated milk, milk powder, condensed milk, fermented milk ( A yogurt, etc., a lactic acid bacteria beverage, a reconstituted cheese, an ice cream, a milk fat, and a milk protein concentrate (MPC), and such a concentrate, such a dried product, such a frozen product, and the like. In this case, the raw material and foods and drinks containing the whey protein may be obtained and used as a commercial product, and may be prepared and used by themselves.

The WPH of the present invention can be prepared by hydrolyzing the above whey protein once or more. The means and/or method are not particularly limited. For example, the whey protein may be partially hydrolyzed in the step of separating or purifying the whey protein from milk or whey, and then further subjected to additional hydrolysis to prepare. The hydrolysis can be carried out in accordance with a hydrolysis method known to the public, but a hydrolysis method using a proteolytic enzyme is preferred.

The proteolytic enzyme is not particularly limited as long as it is a hydrolyzable whey protein and produces a WPH which exerts the effects of the present invention. For example, the type, reaction conditions, and the like can be arbitrarily set with reference to the description of International Publication No. 2007/123200. For example, a proteolytic enzyme derived from a microorganism belonging to the genus Bacillus or Aspergillus; a papaya enzyme derived from papaya, a pineapple enzyme derived from pineapple, a kiwifruit protease derived from kiwifruit, and the like Decomposing enzymes; trypsin, trypsin, pepsin, chymotrypsin and other proteolytic enzymes from animals. These may be used alone or in combination of two or more. Further, commercially available products can be used as such proteolytic enzymes. Commercially available proteolytic enzymes include, for example, Protease M. Amano (Tano Co., Ltd.), Protease N. Amano (Tano Co., Ltd.), Protease P. Amano (Tano Co., Ltd.), Protease A, Amano (Tano Co., Ltd.), Trypsin (Novo Corporation) ), pepsin (Wako Pure Chemical Industries, Ltd.), Umamizyme (Tanozyme Co., Ltd.), Flavourzyme (Novo Corporation), etc., but are not limited thereto. According to an ideal aspect of the present invention, the proteolytic enzyme used to hydrolyze whey protein is selected from the group consisting of a proteolytic enzyme from the genus Bacillus, a proteolytic enzyme from the genus Fusarium, trypsin, pepsin and Flavourzyme (Novo). One or more of the groups.

The WPH of the present invention can prepare a hydrolyzate rich in at least one dipeptide selected from the group consisting of Ile-Leu, Leu-Leu, and Val-Leu by using a proteolytic enzyme that hydrolyzes whey protein. For example, in the case of hydrolyzing whey protein to produce a dipeptide of Ile-Leu, it is preferred to combine the protease derived from Bacillus and the protease derived from the genus Fusarium to hydrolyze whey protein. Further, in the case of hydrolyzing whey protein to produce a Leu-Leu dipeptide, it is preferred to react the whey protein with pepsin and then with a protease derived from the genus Fusarium. In addition, hydrolysis of whey protein by this method sometimes produces dipeptides of Leu-Val and Gly-Leu. Further, for hydrolyzing whey protein to produce a dipeptide of Val-Leu, it is preferred to use a protease derived from the genus Fusarium to hydrolyze whey protein. Hydrolysis of whey protein by this method sometimes produces dipeptides of Ile-Val, Ile-Ile and Leu-Ile. Further, in addition to the hydrolysis of the whey protein to produce the Ala-Leu dipeptide, it is preferred to react the whey protein with trypsin and then with the protease derived from the genus Bacillus.

The reaction temperature required to hydrolyze the whey protein by the proteolytic enzyme to form the desired dipeptide is preferably 20 to 80 ° C, preferably 25 to 75 ° C, more preferably 30 to 70 ° C, and 35 to 65. °C is better, 40~60°C is better, and 45~55°C is especially good. If the reaction temperature is above 20 ° C, the whey protein can be efficiently hydrolyzed by the proteolytic enzyme. Further, when the reaction temperature is 80 ° C or lower, the proteolytic enzyme can be inhibited from being deactivated by denaturation. The reaction temperature may be set to a certain temperature (one stage) depending on the optimum temperature of the proteolytic enzyme to be used, or may be set to two or more different temperatures (two stages or more). In the case where the reaction temperature of two or more stages is set, the order is arbitrary.

The reaction time required to hydrolyze the whey protein by the proteolytic enzyme to form the desired dipeptide is not limited, but it is preferably 2 to 48 hours, preferably 2 to 36 hours, and more preferably 2 to 24 hours. 2 to 18 hours is better, 2 to 14 hours is better, 4 to 12 hours is better, 6 to 10 hours is better, and 7 to 9 hours is particularly good. By setting the reaction time of the proteolytic enzyme to 2 hours or longer, more whey protein can be hydrolyzed. In order to efficiently hydrolyze the whey protein, it is preferred to control the reaction time within 48 hours. Further, when the reaction time is set to two or more stages, the reaction temperature of two or more stages can be arbitrarily set within the range of the reaction time.

The amount of the protease to be added to the desired dipeptide by hydrolyzing the whey protein with the proteolytic enzyme is not particularly limited as long as it can produce WPH which exerts the effect of the present invention. For example, it is preferably 0.5 to 10 g, more preferably 0.7 to 5 g, more preferably 0.8 to 4 g, and particularly preferably 1 to 2 g, based on 100 g of whey protein.

The pH of the reaction required to hydrolyze the whey protein by the proteolytic enzyme to form the desired dipeptide is preferably a pH range in which the proteolytic enzyme is used for the reaction, and the pH is more suitable at or near the optimum pH. . For example, in the case of combining proteolytic enzymes from Bacillus and proteolytic enzymes from the genus Fusarium, pH 5 to 9 is preferred, pH 5.5 to 8.5 is preferred, and pH 6 to 8 is preferred. It is especially good at pH 6.5~7.5.

The pH of the reaction may be set to a certain pH condition (one stage) depending on the optimum pH of the proteolytic enzyme to be used, or may be set to two or more different pH conditions (two stages or more). In the case where pH conditions of two or more stages are set, the order is arbitrary.

As described above, the WPH of the present invention is characterized by comprising at least one dipeptide selected from the group consisting of Ile-Leu, Leu-Leu and Val-Leu. It is preferred to include two or more of these dipeptides, and it is preferred to include three such dipeptides. Examples of the combination of two or more types include Ile-Leu and Leu-Leu, Ile-Leu and Val-Leu, Leu-Leu and Val-Leu or Ile-Leu, and Leu-Leu and Val-Leu. example. The WPH of the present invention can be used as an index containing the specific dipeptide described above.

The dipeptides contained in the WPH of the present invention can be measured by LC/MS and LC/MS/MS. Specifically, it can be measured by the method described in Production Example 1 to be described later. In addition to the hydrolysis reaction conditions as described above, the WPH of the present invention containing these dipeptides can be produced by referring to the examples described in Production Examples 1 to 11 which will be described later.

The content (mg) of these dipeptides per 1 g of WPH can be set as follows. Ile-Leu: 0.1~25mg/gWPH, preferably 0.5~15mg/gWPH, and 1~10mg/gWPH is preferred. Leu-Leu: 0.5~50mg/gWPH, preferably 1~25mg/gWPH, and 2~10mg/gWPH is preferred. Val-Leu: 0.5~50mg/gWPH, preferably 1~25mg/gWPH, and 2~10mg/gWPH is preferred.

The WPH of the present invention may contain at least one of the above dipeptides in a ratio of the protein content per gram of WPH described below. Ile-Leu: 0.09~22.5mg/g protein, preferably 0.45~13.5mg/g protein, and 0.9~9mg/g protein is preferred. Leu-Leu: 0.45~45mg/g protein, preferably 0.9~22.5mg/g protein, and 1.8~9mg/g protein is preferred. Val-Leu: 0.45~45mg/g protein, preferably 0.9~22.5mg/g protein, and 1.8~9mg/g protein is preferred.

Here, the "protein content of WPH" represents the total amount of the amino acid contained in the WPH, and the amino acid which constitutes the protein and peptide contained in the WPH. This protein content can be calculated by the Kjeldahl method. Actually, in the case of the Kjeldahl method, the nitrogen contained in the target WPH can be measured, and this value is calculated by multiplying the value by a nitrogen-protein conversion factor (usually 6.25). For example, since WPH of the present invention is obtained by hydrolyzing whey protein, the protein content of WPH calculated by the above method is theoretically the same as the protein content of whey protein before hydrolysis.

Further, in addition to the above, the WPH of the present invention comprises at least one dipeptide selected from the group consisting of Ile-Leu, Leu-Leu, and Val-Leu, preferably two or more dipeptides, and three or more dipeptides. Further preferably, it may further comprise at least one type of dipeptide selected from the group consisting of Ile-Val, Ile-Ile, Leu-Val, and Leu-Ile. It is preferred to include two or more of these dipeptides, preferably three or more of these dipeptides, and more preferably four of these dipeptides. Examples of the combination of two or more types include Ile-Val and Ile-Ile, Ile-Val and Leu-Val, Ile-Val and Leu-Ile, Ile-Ile and Leu-Val, and Ile-Ile. Leu-Ile, Leu-Val and Leu-Ile, Ile-Val and Ile-Ile and Leu-Val, Ile-Val and Ile-Ile and Leu-Val, Ile-Ile and Leu-Val and Lue-Ile, Ile- Val and Ile-Ile with Leu-Val and Leu-Ile. The WPH of the present invention can be used as an indicator containing the specific dipeptide described above.

The dipeptides contained in the WPH of the present invention can also be measured by LC/MS and LC/MS/MS. The content of each WPH of these dipeptides can be set as follows. Ile-Val: 0.1~10mg/gWPH, preferably 0.2~5mg/gWPH, preferably 0.5~1.5mg/gWPH, Ile-Ile: 0.05~10mg/gWPH, preferably 0.1~5mg/gWPH, 0.2~1mg/gWPH is preferred Leu-Val: 0.1~10mg/gWPH, preferably 0.2~5mg/gWPH, 0.5~1.5mg/gWPH is preferred Leu-Ile: 0.01~10mg/gWPH, 0.02~ 5 mg/g WPH is preferred, and 0.05 to 0.2 mg/g WPH is preferred.

The WPH of the present invention may contain at least one of the above dipeptides in a ratio of the protein content per gram of WPH described below. The meaning of "protein content of WPH" is as described above. Ile-Val: 0.09~9mg/g protein, preferably 0.18~4.5mg/g protein, 0.45~1.35mg/g protein is preferred Ile-Ile: 0.045~9mg/g protein, 0.09~4.5mg/ g protein is better, 0.18~0.9mg/g protein is preferred Leu-Val: 0.09~9mg/g protein, preferably 0.18~4.5mg/g protein, and 0.45~1.35mg/g protein is preferred Leu -Ile: 0.0090-1mg/g protein, preferably 0.018~4.5mg/g protein, preferably 0.045~0.2mg/g protein.

The WPH of the present invention includes the total amount of the dipeptides as Ile-Leu, Leu-Leu, and Val-Leu, and is preferably 1.1 to 125 mg/g WPH per 1 g of WPH, preferably 2.5 to 65 mg/g WPH, and 5 ~30 mg/g WPH is a preferred ratio inclusion. The total amount of such dipeptides contained in the WPH is converted to a protein content of 0.99 to 112.5 mg/g protein per 1 g of WPH, preferably 2.25 to 58.5 mg/g protein, and 4.5 to 27 mg/g protein. good.

Further, the WPH of the present invention includes the total amount of the dipeptides as Ile-Val, Ile-Ile, Leu-Val, and Leu-Ile, and is 0.26 to 40 mg/g WPH per 1 g of WPH, and 0.52 to 20 mg/g WPH. Good, including 1.25~4.2mg/gWPH as the preferred ratio. The dipeptide contained in the WPH is preferably 0.234 to 36 mg/g protein per 1 g of WPH, preferably 0.468 to 18 mg/g protein, and preferably 1.125 to 3.8 mg/g protein.

The WPH of the present invention is liquid when it is just prepared, but can be freeze-preserved, dried (spray-dried, freeze-dried), concentrated, and heat-sterilized (plate type heat sterilization, tube type heat sterilization, intermittent heating) in order to improve the storage stability. Indirect heat sterilization such as sterilization, electric heating sterilization, microwave heat sterilization, steam heating, steam infusion, steam infusion, etc., or non-heat sterilization (light irradiation sterilization, radiation irradiation sterilization, high voltage pulse sterilization). Among them, from the viewpoints of preservation stability and ease of handling, it is preferred to dry-treat (spray-dry, freeze-dry) WPH after preparation and prepare a dry solid form.

As shown in Test Examples 1 and 2 described later, the composition of the present invention containing WPH can be administered to a mammal (including humans and non-human animals) by ingesting or ingesting it and giving exercise load to the body, particularly muscles. The amount of BDNF is increased (refer to Figures 1 and 2). Moreover, the amount of increase is higher in the body, particularly the amount of BDNF in the muscle, which is increased in the case of not administering to the mammal or ingesting the composition of the present invention but giving exercise load. Therefore, the composition of the present invention containing WPH is useful as a composition for promoting an increase in the amount of BDNF in the body, particularly in the muscle (a composition for promoting an increase in the amount of BDNF in the body, particularly in the muscle). Furthermore, the composition for promoting an increase in the amount of BDNF can be used to increase or ingest the mammal by pre-exercise or exercise, thereby improving the body after exercise, particularly intramuscular BDNF. The amount is better. Herein, "promoting an increase in the amount of BDNF" means the amount of BDNF in the body after exercise (intake) of the composition of the present invention in the case of mammalian pre- or post-exercise exercise, particularly intramuscular BDNF, and This amount is higher in the case where the composition of the present invention is not administered (ingested). Here, "when the composition of the present invention is not administered (ingested)", the case where the composition of the present invention is not administered (ingested) is not given an exercise load, and the present invention is not administered (ingested). The composition is given to both the exercise load. More specifically, it is represented by the administration of the composition of the present invention before or during exercise to promote intramuscular in the case of not administering (ingesting) the composition of the present invention but giving exercise load The amount of BDNF is increased, and the amount of BDNF in the muscle is increased more than in the case of not administering (uptake) the composition of the present invention but giving an exercise load.

In addition, in the present invention, the term "exercise" may be caused by contraction of skeletal muscle with more energy than rest, that is, a burden on the muscle. As long as there is no particular limitation within this range, specifically, it can be carried out to maintain physical strength, such as walking, brisk walking, jogging, running, dancing, water sports, skiing, cycling, gymnastics, For example, yoga, stretching, weight training, compression training, swimming, mountain climbing, rock climbing, various sports (such as ball games, badminton, ballet, etc.).

Further, as shown in Test Example 2 described later, the composition of the present invention containing WPH can be made into the body, particularly the amount of BDNF in the body, by administering or ingesting it to the mammal even without the exercise load. Increase (see Figure 3). In this case, the "increased amount of BDNF" means the amount of BDNF in the body, especially the amount of BDNF in the muscle, without giving exercise load but administering (uptake) the composition of the present invention to the mammal, and not giving exercise load and not This amount is higher in the case of administration (ingestion) of the composition of the present invention. Therefore, even in the case where no exercise load is given, the composition of the present invention containing WPH is useful only as a composition for promoting an increase in the amount of BDNF in the body, particularly by ingestion into the body.

Here, the "injection or ingestion" is not limited as long as it is placed in a mammal (including humans and non-human animals), but may be exemplified by oral administration or ingestion, enteral administration, and stomach cramps. It is preferred to digest and absorb in the intestine, and may be appropriately selected depending on the state and use of the mammal to be targeted. Oral ingestion is preferred. Therefore, the composition of the present invention has a form for oral administration or ingestion, a form for enteral administration, and a form for gastric fistula, and can be prepared in such a form.

As described above, BDNF is known to be one of the neurotrophic factors involved in the production, growth, maintenance, and regeneration of nerve cells, and enhances the expression of memory and learning (Non-Patent Document 3). Therefore, the composition of the present invention can be used as a component for assisting memory or learning performance improvement, and for those who need to improve memory, learning ability, or learning efficiency (for example, candidates, memory, learning ability, learning efficiency, etc.) Ingest or give it.

Further, it is known that the amount of BDNF expression in the brain of patients with depression and Alzheimer's type dementia is reduced (Non-Patent Documents 8 and 9). Therefore, the composition of the present invention can be used as a composition for improving cognitive function, for suppressing cognitive deterioration or for enhancing cognitive function, and for dementia, especially for Alzheimer's dementia or possibly The sick person ingests or administers it. Here, the "cognitive function" means that the correct understanding of things and the proper implementation of the required functions include memory, speech ability, judgment (including orientation, attention), computational power, and/or execution. And the possible patients with dementia also include those with mild cognitive impairment. In addition, an elderly person over the age of 65 is considered to have dementia or mild cognitive impairment in every 4 people. Therefore, in the present invention, an elderly person over 65 years old can also be included in a possible patient with dementia. . Further, the composition of the present invention can be used as a composition for ameliorating depression, in which a patient suffering from depression (including a new type of depression) or a patient with a depressed state or a tendency to have depression is ingested or administered.

Further, it has been reported that by administering BDNF intracerebroventricularly, subcutaneously or intraperitoneally, the increase in body weight is suppressed and the glucose metabolism is improved (Non-Patent Document 10). Therefore, the composition of the present invention can be used as a composition for inhibiting or ameliorating obesity, which is ingested or administered to an obese person, an obese person or a patient who is likely to have a disease. Further, the composition of the present invention can be used as a composition for suppressing a decrease in sugar metabolism or for improving glucose metabolism, and allowing a diabetic patient, a diabetic patient or a patient thereof to ingest or administer it.

The composition of the present invention is limited to the effect of the present invention, and other food materials and/or food additive materials may be added in addition to the WPH of the present invention. For example, in order to impart preference to the composition of the present invention, a sweetener (sucrose, liquid sugar, fructose, maltose, tri- warm sugar, etc.), a sugar alcohol (erythritol, etc.), a high-sensitivity sweetener may be added ( Stevia, aspartame, sucralose, acesulfame potassium, etc.), sour materials, spices, fruit juices, vegetable juices, polysaccharides, emulsifiers, flavoring agents, flavoring agents, etc. In addition, it is also possible to add minerals (calcium, iron, manganese, magnesium, zinc, etc.), vitamins, functional materials, and probiotic lactic acid bacteria for the purpose of imparting additional functions (nutritional functions and physiological functions) to the composition of the present invention. Etc. to form the composition of the present invention.

The composition of the present invention can be formed into a container for the purpose of being provided on the market. As a form of the container, a state in which the composition of the present invention is prepared into a solid form and then filled into a container can be mentioned. Examples of the form of the solid can be powders and granules. As the container in this case, paper, bags, boxes, cans, bottles, and the like can be used as known to the public. The amount of the container at this time is not particularly limited, and is, for example, 1 to 5000 g, 2 to 4000 g, 3 to 3000 g, 4 to 2000 g, 4 to 1000 g, 4 to 500 g, 4 to 250 g, 4 to 200 g, 4 to 100 g, and 4 ~50g, 4~30g. Among them, in the case of family wear (multiple people taking), it is 300~5000g, 300~4000g, 300~3000g, 300~2000g, 300~1500g, 300~1400g, 300~1300g, 300~1200g, 300~1100g, In the case of personal wear (individual use), it is 1~500g, 2~250g, 3~200g, 4~100g, 4~50g, 4~40g, 4~30g, 5~30g, 10~30g, 15~30g .

Further, the composition of the present invention can be formed into a container-packed beverage for the purpose of being provided on the market. Here, as long as the container is a container that can be directly consumed after the beverage is filled, a paper container, a soft bag, a PET bottle, a can, a bottle, or the like can be used as a public. The beverage can be prepared by dispersing and dissolving the above powder or granules in drinking water. The amount of the contents of the container is not particularly limited, and is, for example, 1 to 5000 g, 2 to 4000 g, 3 to 3000 g, 4 to 2000 g, 5 to 1000 g, 6 to 800 g, 7 to 1300 g, 80 to 1200 g, and 90 to 1100 g. In the case of a family pack (multiple people), for example, 300 to 5000 g, 300 to 4000 g, 300 to 3000 g, 300 to 2000 g, 300 to 1500 g, 300 to 1400 g, 300 to 1300 g, 300 to 1200 g, and 300 to 1100 g. In the case of personal wear (individual drinking), it is 10~500g, 20~500g, 30~500g, 40~500g, 50~500g, 60~500g, 70~500g, 80~500g, 90~500g.

According to one embodiment of the present invention, the composition of the present invention can also be ingested (administered) in such a manner that the total protein content of WPH is 0.1 g or more (preferably 1 g or more) per administration (uptake). good. The total protein content at the time of administration (ingestion) is preferably from 0.1 to 40 g, preferably from 0.1 to 30 g, and from 1 to 30 g, for better intake (administration).

The "one-time administration (ingestion)" represents the amount of each state required to promote an increase in the amount of BDNF in the body. For example, in the case where a series of exercise is performed within a predetermined time of one day, the total amount of the dose or the amount of the injection or ingestion before or during the exercise is taken in several times (in one breath) and divided into several intakes. The amount of this administration (ingestion) divided into several times is regarded as "one-time administration (ingestion)".

From the viewpoint of promoting exercise to increase the amount of BDNF, the composition of the present invention is administered (for ingestion) to a subject (mammal) in which an increase in the body, particularly the amount of BDNF in the muscle, is required before or during exercise. . It can be 15~240 minutes before exercise, 30~180 minutes before exercise, and 45~135 minutes before exercise. In addition, in the case of no exercise, although it can be administered at any time (for ingestion), it can be obtained because it is easy to absorb when it is relatively fasted before meals or between meals. Effect.

According to other embodiments of the present invention, for example, the composition of the present invention can also be provided in a unit package form for each administration (ingestion). The composition of the present invention having the unit package form is preferably prepared by modulating the WPH in such a manner that the total protein content is 0.1 g or more (preferably 1 g or more) in this unit. In addition, it is preferable to prepare the above-mentioned WPH in such a manner that the total amount of protein is 40 g or less (preferably 30 g or less) as a single administration (intake) amount. That is, the content of the WPH of the present invention per unit package is preferably 0.1 to 40 g, preferably 0.1 to 30 g, and preferably 1 to 30 g, as a total amount of the protein (ingestion). In this case, the "unit packaging form for each injection (intake)" means that the amount of each injection (intake) is a predetermined form, for example, as a food or drink that can be taken orally in a certain amount, not only It represents the form of general food, and also represents the form of beverages (drinks, etc.), health supplement foods, health functional foods, nutritional supplements, and the like. "The unit packaging form for each injection (intake)", for example, a liquid beverage, a gel, a paste, a paste, a powder, a granule, a capsule, or a solid food. In the case of a metal can, a glass bottle (bottle, etc.), a plastic container (such as a PET bottle), a packaging, a bag, a film container, a paper box, or the like, a specific amount (amount) can be specified. Or, it is possible to display the amount of each administration (intake) in a packaging container or a web page to specify a specific amount. Since WPH is used as an active ingredient in the composition of the present invention, it is less likely to gel due to heating than whey protein. Therefore, appropriate physical properties can be maintained in the production steps of beverages, jelly, and the like. Further, since the composition of the present invention exerts the effects of the present invention even when ingested in a small amount, all forms are easily ingested and swallowed, regardless of whether it is a block of a large size or a mini canned beverage that can be ingested. It is convenient for children and the elderly.

The composition of the present invention can be provided in the form of a composition in which a plurality of components are blended. The form of the composition of the present invention is not particularly limited as long as it exhibits the effects of the present invention, but may be, for example, a solid (powder, granule, capsule, block, etc.), liquid, gel, or paste. The form of the paste or the like is preferably in the form of a powder. The composition includes a food composition and a pharmaceutical composition.

According to a preferred aspect of the present invention, there is provided a food or drink for promoting an increase in the amount of BDNF, which comprises WPH as an active ingredient. In the present invention, the form of the food is not particularly limited as long as it can be orally ingested, such as a solution, a suspension, an emulsion, a powder, or a solid molded product. Specifically, for example, instant noodles, sterilized food, canned food, microwave food, instant soup. Instant foods such as miso soups, freeze-dried foods; beverages such as refreshing drinks, fruit drinks, vegetable drinks, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, alcoholic beverages, etc.; bread, Italy Flour products such as noodles, noodles, cake mixes, bread flours, etc.; candies, milk candy, chewing gum, chocolate, sweet biscuits, biscuits, cakes, pies, snacks, salty biscuits, Japanese snacks, dessert snacks, etc.; Seasonings such as seasonings, tomato processing seasonings, flavoring seasonings, conditioning powders, sauces, salad dressings, squid soy sauces, curry and stew seasonings; processing oils, creams, margarines, Milk and fats such as cannabis; dairy products, yogurt, lactic acid bacteria beverages, ice creams, milk fats, etc.; agricultural products such as canned food, jam, peel sauce, cereals, etc.; frozen foods. Dairy products are preferred for dairy products, and milk beverages and lactic acid bacteria beverages are preferred.

In addition, foods and drinks include health foods, functional foods, nutritional supplements, functional foods, specific health foods, foods for the sick, milk powder for infants, milk powder for pregnant women or breastfeeding women.

The food and beverage of the present invention may also be accompanied by a statement about "increasing the amount of BDNF in the body", "promoting an increase in the amount of BDNF in the body", and "inducing an increase in the amount of BDNF in the body". Furthermore, the above-mentioned term "in vivo" also includes the meaning of "intramuscular".

According to another aspect of the present invention, there is provided a method of promoting an increase in the amount of BDNF in the body, particularly in the muscle, comprising subjecting a mammal (including a human or a non-human animal) to ingestion or administering WPH thereto. The method for promoting an increase in the amount of BDNF of the present invention can be carried out in accordance with the description of the composition of the present invention.

Here, in the method for promoting an increase in the amount of BDNF of the present invention, it can be used in mammals and samples (muscle tissues, muscle cells) derived therefrom, and the purpose can be either therapeutic use or non-therapeutic use. Herein, the term "non-therapeutic" means that the human is not subjected to surgery, treatment or diagnostic behavior (ie, medical treatment for humans), specifically, a person who does not include a doctor or is instructed by a physician to perform surgery or treatment on a human being. Or a method of diagnosis. [Examples]

Hereinafter, the present invention will be described in more detail by way of production examples and experimental examples. However, these production examples and experimental examples are part of examples for explaining the present invention, and the present invention is not limited thereto. Further, the following production examples and experimental examples were carried out under the conditions of room temperature (25 ± 5 ° C) and atmospheric pressure unless otherwise stated. In addition, the protein content contained in the test sample was determined using the Kjeldahl method.

Production Example 1 (1) Production of whey protein hydrolyzate (dry powder) 50 g of a commercially available whey protein concentrate (hereinafter abbreviated as "WPC") was dissolved in 1 L of water to prepare an aqueous WPC solution, and a pH adjuster was used. It was adjusted to pH 7. 500 mg of a protease derived from Bacillus (Protease M. Amano, Amano Co., Ltd.) and 500 mg of a protease derived from Fusarium (Protease N. Amano, Amano Co., Ltd.) were added thereto, and treatment was carried out at 50 ° C for 8 hours, thereby hydrolyzing the milk. Clear protein. After heating to deactivate the protease, it was freeze-dried and pulverized, and this was taken as whey protein hydrolyzate (WPH) (dry powder).

(2) Component analysis The whey protein hydrolyzate (dry powder) obtained above was diluted and dissolved to 1000 times (volume ratio) in an aqueous solution of trifluoroacetic acid having a concentration of 0.1 (w/w), and tested under the following conditions. LC/MS/MS to determine the dipeptides contained in WPH (Ile-Leu, Leu-Leu, Val-Leu, Ile-Asn, Leu-Ala, Leu-Glu, Ala-Leu, and Lys-Ile) Amount (unit mg/g protein: converted in solid matter, the same applies hereinafter).

[Analytical conditions for LC/MS/MS] Column: ACQUITY UPLC BEHC18 Column, 130Å, 1.7mm, 2.1 ́50mm (Waters) Mobile phase: Liquid A: Aqueous solution B containing 0.1 (w/w)% trifluoroacetic acid : The ratio of the A liquid and the B liquid in the mobile phase containing 0.1 (w/w)% trifluoroacetic acid in the mobile phase begins to flow → 0 points: 100% v/v% of liquid A (0 v/v% of liquid B) 0 points → 9 points: A liquid 100v / v% → A liquid 60v / v% - B liquid 40v / v% 9 points → 10 points: A liquid 60v / v% - B liquid 40v / v% → A liquid 20v / v% - B liquid 80v/v% 10 points→11 points: A liquid 20v/v%-B liquid 80v/v%→A liquid 100v/v% (B liquid 0v/v%) Column temperature: 40°C Flow rate: 0.3mL /Minute.

[MS/MS condition] ESI positive ion mode Capillary voltage: 3000V Source temperature: 120 °C Desolvation temperature: 400 °C Desolvation gas flow rate: 850 L / hour Cone gas flow rate: 50 L / hour Cone voltage: 25 V Collision voltage: 15 mV.

The content of the dipeptide contained in the WPH (dry powder) prepared in the above Production Example 1 is shown in Table 1.

Production Example 2 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of a protease derived from Bacillus (Protease M. Amano, Amano Co., Ltd.) was added thereto, and treatment at 50 ° C for 8 hours was carried out to hydrolyze the whey protein in the WPC aqueous solution. Then, the protease was inactivated, and then freeze-dried and powdered, and this was regarded as WPH (dry powder) of Production Example 2.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 2.

Production Example 3 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of a protease derived from the genus Fusarium (Protease N. Amano, Amano Co., Ltd.) was added thereto, and treatment at 50 ° C for 8 hours was carried out to hydrolyze the whey protein in the WPC aqueous solution. Subsequently, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 3.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 3.

Production Example 4 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of trypsin (Novo) was added thereto, and after reacting at 37 ° C for 4 hours, 500 mg of a protease derived from Bacillus (Protease M. Amano, Amano Co., Ltd.) was added, and further reacted at 50 ° C for 4 hours to hydrolyze WPC. Whey protein in aqueous solution. Then, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 4.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 4.

Production Example 5 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of trypsin (Novo) was added thereto, and after reacting at 37 ° C for 4 hours, 500 mg of a protease derived from Fusarium (Protease N. Amano, Amano Co., Ltd.) was added, and further reacted at 50 ° C for 4 hours to hydrolyze WPC. Whey protein in aqueous solution. Then, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 5.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 5.

Production Example 6 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of pepsin (Wako Pure Chemical Industries, Ltd.) was added thereto, and after reacting at 37 ° C for 4 hours, 500 mg of a protease derived from Bacillus (Protease M. Amano, Amano Co., Ltd.) was added, and further reacted at 50 ° C for 4 hours to The whey protein in the WPC aqueous solution is hydrolyzed. Then, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 6.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 6.

Production Example 7 50 g of a commercially available WPC (the same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of pepsin (Wako Pure Chemical Industries, Ltd.) was added thereto, and after reacting at 37 ° C for 4 hours, 500 mg of a protease derived from Fusarium (Protease N. Amano, Amano Enzyme Co., Ltd.) was added, and further reacted at 50 ° C for 4 hours to Hydrolyzing whey protein in aqueous whey protein solution. Then, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 7.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 7.

Production Example 8 50 g of a commercially available WPC (the same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of Flavourzyme (Novo Corporation) was added thereto, and reacted at 50 ° C for 6 hours to hydrolyze whey protein in the aqueous solution of whey protein. Then, the protease was inactivated, and then freeze-dried and powdered, and this was regarded as WPH (dry powder) of Production Example 8.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 8.

Production Example 9 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of a protease derived from Fusarium (Umamizyme, Amano Co., Ltd.) was added thereto, and reacted at 50 ° C for 6 hours to hydrolyze whey protein in the WPC aqueous solution. Then, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 9.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 9.

Production Example 10 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of a protease derived from Fusarium (Protease A Amano, Amano Co., Ltd.) was added thereto, and reacted at 50 ° C for 6 hours to hydrolyze whey protein in the WPC aqueous solution. Then, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 10.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 10.

Production Example 11 50 g of a commercially available WPC (same as WPC used in Production Example 1) was dissolved in 1 L of water to prepare a WPC aqueous solution, which was adjusted to pH 7 with a pH adjuster. 500 mg of a protease derived from Fusarium (Protease P. Amano, Amano Co., Ltd.) was added thereto, and reacted at 50 ° C for 6 hours to hydrolyze whey protein in the aqueous solution of whey protein. Then, the protease was inactivated, and then freeze-dried and pulverized, and this was regarded as WPH (dry powder) of Production Example 11.

The amount of the dipeptide contained in the above-mentioned prepared WPH (dry powder) was measured in accordance with the analysis method described in the item (2) of Production Example 1. The results are shown in Table 11.

Test Example 1 This test was conducted to evaluate the effect of the combination of the uptake and exercise of the whey protein hydrolyzate as a milk material on cognitive function by using the BDNF concentration in the muscle as an index. In the test, rats were used to measure and compare the conditions in which the whey protein hydrolyzate was ingested and given an exercise load, and the WPH was not taken but the exercise load was given, and the whey protein hydrolyzate was not ingested. Concentration of BDNF contained in skeletal muscle with exercise load.

(1) Test method (1-1) Preparation of test sample (test food) In the test food of Test Example 1, WPH (compared with WPC used in Production Example 1) commercially available by protease hydrolysis was used ( Dry powder). Further, the solid content per 100% by mass of the WPH contains about 89.3% by mass of protein. The WPH was dissolved in distilled water in such a manner that the solid concentration reached 10 w/v%, and this was used as a test sample (test food).

The amount of the dipeptide (mg/g WPH, mg/g protein) contained in the prepared WPH (dry powder) was measured according to the analysis method described in the item (2) of Production Example 1. The results are shown in Table 12.

(1-2) Preparation of test animals Male SD rats of 8 weeks old were used as test animals. Pre-feeding was carried out for 4 to 5 days in advance, and after the end of the feeding, the lower right leg was depilated under isoflurane anesthesia at the latest before the test. In addition, a 16-hour fast was started on the day before the test.

On the day of the test, the population was divided as follows by equal weight (about 290 to 320 g). [Experimental group (3 groups in total)] (a) "Double-peptide non-administered/non-exercised group" (n=6): The test sample is not administered without giving exercise load (b) "Dipeptide is not administered" /Motion group" (n=6): The exercise load was 50 times, and the test sample was not administered. (c) "Dipeptide administration/exercise group" (n=3): The exercise load was 50 times, and the test sample was administered.

(1-3) Test A single oral administration of 10 ml/kg body weight (WPH 1.0 g/kg body weight containing dipeptide) to the test animals of "dipeptide administration/exercise group" among the above-mentioned groups of the test animals. ) to the test sample (test food) prepared as described above. Thirty minutes after the administration, the right hind limb exercise load was forcibly administered by electrical stimulation under isoflurane anesthesia (details will be described later). Furthermore, the test sample (test food) was replaced with distilled water, and the test animals of the "bipeptide non-administered/exercise group" were orally administered at a ratio of 10 ml/kg body weight, and 30 minutes after the administration. The right hind limb exercise load was forcibly administered by electrical stimulation under isoflurane anesthesia in the same manner as described above. In addition, the test sample (test food) was replaced with distilled water, and the test animals of "dipeptide non-administered/non-exercised group" were orally administered at a ratio of 10 ml/kg body weight, and then under isoflurane anesthesia. Let it rest.

The test animals of "dipeptide administration/exercise group" and "dipeptide non-administration/exercise group" were cast 3 hours after the end of exercise load, and the test animals of "bipeptide not administered/not exercised group" were cast. After 3 hours and 30 minutes of distilled water, whole blood was collected from the large inferior vena cava under isoflurane anesthesia for slaughter.

Thereafter, the lower extremity skeletal muscles are quickly taken and weighed. The skeletal muscle measurement taken was used as the BDNF concentration (ng/g) of the cognitive function evaluation index.

[Sports load method] Rats were fixed in a prone manner on a dedicated fixation table of a device for giving electrical stimulation to exercise load under isoflurane anesthesia. Press the knee that secures the right hind limb and secure the ankle to the foot pedal. The skin electrode is attached to the calf and forced to move by electrical stimulation. The mandatory exercise load is a step of electrical stimulation every 8 seconds, repeated 5 times as a group, and the exercise group is subjected to 10 groups (a total of 50 electrical stimulations). Furthermore, the pattern of the exercise load is set to the isometric contraction load (ISO).

(2) Results The results are shown in Fig. 1. The comparison of (b) "bipeptide non-administration/exercise group" and (a) "bipeptide non-administered/non-exercised group" in Fig. 1 confirmed that the concentration of BDNF in the muscle increased due to exercise. Moreover, the comparison of (b) "bipeptide non-administered/exercise group" and (c) "dipeptide administration/sports group" confirmed that the concentration of BDNF in the muscle was increased by exercise plus the sample of ingestion (WPH). Rise more in the case of only sports. In addition, the comparison of (a) "bipeptide non-administered/non-exercised group" and (c) "dual peptide administration/exercise group" confirmed that the concentration of BDNF in the muscle was higher than that of the sample due to ingestion of the test sample and exercise. Significantly elevated without exercise (Student's t-test: P < 0.05).

Thus, the WPH containing the dipeptide which is the object of the present invention is exhibited, and the effect of promoting the production of BDNF in skeletal muscle is exhibited by combining internal administration and exercise. In addition, BDNF is known to pass the blood-brain barrier (Pardridge WM et al. Pharm Res. 11(5): 738-46 (1994). Therefore, it is speculated that BDNF produced in muscle will be transferred to the brain through the blood-brain barrier, and It contributes to the suppression of deterioration of cognitive function. That is, by ingesting the whey protein hydrolyzate of the present invention and exercising, an effect of suppressing deterioration of cognitive function can be expected.

Test Example 2 This test was conducted to evaluate the effect of WPH uptake on cognitive function by using BDNF concentration in muscle as an index. In the test, rats were used, and the concentration of BDNF contained in the skeletal muscle in the case where the whey protein hydrolyzate was replaced with water or the whey protein before hydrolysis and ingested with WPH was measured and compared. Further, the concentration of BDNF contained in the skeletal muscle in the case where WPH is taken up by the water or the whey protein before hydrolysis is taken up and the exercise load is given, and the exercise load is given, is also measured and compared, To confirm the impact of using with sports.

(1) Test method (1-1) Preparation of test sample (test food, comparative test food) In the test food of the test example 2, as in Test Example 1, the commercially available WPC was hydrolyzed by protease (with Manufacturing Example 1). WPH (dry powder) obtained by the same WPC used (refer to Table 12). The WPH was dissolved in distilled water in such a manner that the solid concentration reached 10 w/v%, and this was used as a test sample (test food). On the other hand, whey protein before the hydrolysis (commercial WPC, the same as WPC used in Production Example 1) was used. In addition, regardless of the test food (WPH) or the comparative test food (whey protein), the dose was converted into a protein content in an amount of 1.0 g protein/body weight kg.

(1-2) Preparation of test animals Male SD rats of 8 weeks old were used as test animals. Pre-feeding was carried out for 4 to 5 days in advance, and after the end of the feeding, the lower right leg was depilated under isoflurane anesthesia at the latest before the test. In addition, a 16-hour fast was started on the day before the test.

On the day of the test, the population was divided as follows by equal weight (about 290 to 310 g). [Test group (6 groups in total)] <Motion group> (1) "Water injection/sports group" (n=5): 50 times of exercise load, water injection (2) "WPC investment/sports group" ( n=5): exercise load 50 times, administration of 1.0 g protein/body weight kg comparison test food (whey protein) (3) "WPH administration/exercise group" (n=5): exercise load 50 times, administration 1.0 g protein/kg kg of test food (WPH). <No sports group> (i) "Water injection/non-sports group" (n=5): no exercise load, water injection (ii) "WPC cast/no motion group" (n=5): no exercise Load, 1.0 g protein/kg kg comparison test food (whey protein) (iii) "WPH administration/non-exercise group" (n=5): no exercise load, 1.0 g protein/kg kg test food (WPH).

(1-3) Test A single oral administration of water or a test sample to the test animals of the above exercise group was performed, and after 15 minutes, the right lower limb exercise load was forcibly administered by electrical stimulation under isoflurane anesthesia. The exercise load method is as described in Test Example 1. The test animals of the above non-exercise group were orally administered with water or test samples, and 15 minutes later, isoflurane anesthesia was administered simultaneously with the aforementioned exercise group. After the exercise group was 180 minutes after the end of the exercise load, the non-exercise group was subjected to total blood collection by the large inferior vena cava under the isoflurane anesthesia for 180 minutes after the end of anesthesia. Thereafter, the lower extremity skeletal muscles are quickly taken and weighed. The skeletal muscle measurement taken was used as the BDNF concentration (ng/g) of the cognitive function evaluation index.

(2) Results The results of the sports group and the non-moving group are shown in Figures 2 and 3, respectively. As shown in Figure 2, (2) compared with the animals tested in the "WPC/Sports Group" and (1) "Water Injection/Motion Group", the BDNF concentration in the muscle increased slightly, but it was not obvious. difference. In contrast, it was confirmed that (3) the "WPH administered/sports group" test animals and (1) the "water administration/sports group" test animals significantly increased the muscle BDNF concentration. In addition, as shown in Figure 3, the same situation occurs in the non-sports group, (ii) the "WPC cast/non-sports group" test animals and (i) the "water cast/non-sports group" test animals, Although the concentration of BDNF in the muscle increased slightly, there was no significant difference, but relatively, (iii) "WPH administered/non-exercise group" test animals and (i) "water administration/non-exercise group" test animals were confirmed. In contrast, the concentration of BDNF in the muscle is significantly increased.

From the results of the above Test Examples 1 and 2, it was confirmed that a significant increase in the concentration of BDNF in the muscle due to the ingestion of the test sample (WPH containing the dipeptide) occurred regardless of the presence or absence of exercise. This remarkable effect only occurs when the ingestion contains dipeptides (Ile-Leu, Leu-Leu, Val-Leu, Ile-Val, Ile-Ile, Leu-Val, Leu-Ile, Ile-Asn, Leu-Ala, Leu- When WPH of Glu, Ala-Leu or/and Lys-Ile) was observed, no significant effect was observed when whey protein containing no such dipeptide was ingested. That is, it is confirmed that the WPH containing the dipeptide of the present invention is taken up by the combined exercise (for example, before exercise or during exercise), or is not used in combination with exercise but ingests the WPH containing the dipeptide of the present invention (without exercise) Ingestion) promotes BDNF production in skeletal muscle, and the body's content of BDNF increases. As mentioned above, there are reports suggesting that BDNF is effective in inhibiting cognitive deterioration, improving memory and learning performance, inhibiting obesity, or inhibiting decreased glucose metabolism. Therefore, it is considered that the WPH containing the dipeptide of the present invention has an effect of suppressing the deterioration of cognitive function, improving the expression of memory and learning, suppressing obesity, or suppressing the decrease in glucose metabolism by promoting the action of BDNF production in vivo.

[Fig. 1] shows that the test animals (male SD rats) are classified into (a) "dipeptide non-administered/non-exercise group", (b) "dipeptide non-administered/exercise group", and (c) "Dipeptide administration/exercise group", and compare (a) the amount of BDNF (ng/g) in the muscle without ingesting WPH or giving exercise load (mean ± standard error, the same below) (b) the result of the same amount of BDNF in the case where WPH was not taken but the exercise load was given, and (c) the amount of BDNF after the WPH was taken and the exercise load was given. In the figure, * represents a significant difference of p < 0.05 with respect to (a) according to the Dunnett's test. [Fig. 2] Separately, the test animals (male SD rats) are classified into (1) "administered water/sports group", (2) "administered to WPC/sports group", and (3) "administered WPH" /Motion group", and measure (1) the amount of BDNF (ng/g) in the muscle after ingesting water and giving exercise load (mean ± standard error, the same below), (2) ingesting WPC (milk The result of clearing the protein concentrate) and giving the amount of BDNF after the exercise load and (3) taking the WPH and giving the amount of BDNF after the exercise load. In the figure, * represents a significant difference of p < 0.05 with respect to (1) according to the Dunnett test. [Fig. 3] Separately, the test animals (male SD rats) are classified into (i) "administered water/non-sports group", (ii) "invested in WPC/non-sports group", and (iii) "invested". For the WPH/non-exercise group, all the exercise load is not given, and (i) the amount of BDNF (ng/g) after the water is taken (mean ± standard error, the same below), (ii) ingestion The amount of BDNF after WPC and (iii) the result of the same amount of BDNF after ingesting WPH. In the figure, * represents a significant difference of p < 0.05 with respect to (i) according to the Dunnett test.

Claims (13)

A use, which is a use of a whey protein hydrolysate for producing a composition for promoting an increase in the amount of brain-derived neurotrophic factor (BDNF) in a subject, and the whey protein hydrolyzate is selected from the group consisting of Ile - at least one dipeptide group of Leu, Leu-Leu and Val-Leu. The use of claim 1, wherein the whey protein hydrolyzate is a dipeptide comprising Ile-Leu, Leu-Leu and Val-Leu. The use of claim 1 or 2, wherein the whey protein hydrolyzate is further comprising at least one dipeptide selected from the group consisting of Ile-Val, Ile-Ile, Leu-Val, and Leu-Ile. The use of claim 1 or 2, wherein the whey protein hydrolyzate is a dipeptide further comprising Ile-Val, Ile-Ile, Leu-Val and Leu-Ile. The use of any one of claims 1 to 4, wherein the aforementioned subject is a mammal comprising humans before or during exercise. The use of claim 5, wherein the composition is for promoting an increase in the amount of BDNF in the muscle during or after exercise of the subject. The use of any one of claims 1 to 4, wherein the subject is a mammal comprising a human in a non-exercise state. The use of any one of claims 1 to 7, wherein the aforementioned subject is a cognitive degenerative person or a patient thereof. The use of claim 8, wherein the aforementioned composition is for suppressing or improving cognitive function deterioration. The use of any one of claims 1 to 9, wherein the aforementioned subject is an obese person or/and a reduced glucose metabolism, or such a likely patient. The use of claim 10, wherein the composition is for inhibiting or ameliorating obesity, or/and for reducing or improving glucose metabolism. The use according to any one of claims 1 to 11, wherein the composition is composed of a unit packaging type for each administration (intake), and is included as a single administration amount in the unit packaging type (ingestion) The composition of the above whey protein hydrolyzate in an amount of 0.1 to 40 g in terms of total protein. The use of any one of the items 1 to 12, wherein the composition is a food or drink, a health food, a functional food, a nutritional supplement food, a functional food, a specific health food, a patient food, an infant It is better to use milk powder for pregnant children, milk powder for pregnant women or lactating women, or food supplements with a statement to promote exercise to increase the amount of BDNF in the body.