JP7392474B2 - Food composition for improving brain function - Google Patents

Description

The present invention relates to food compositions for improving brain function.

In recent years, there is no stopping the increase in the proportion of the elderly population worldwide. The Statistics Bureau of Japan's Ministry of Internal Affairs and Communications predicts that the proportion of the world's population aged 65 and over will more than double to 16% in 2050, which was 7.6% in 2010. Furthermore, with this global transition to an aging society, the number of patients with brain function disorders is expected to increase. Regarding Alzheimer's disease, which is a brain functional disease, the International Alzheimer's Association's "World Alzheimer's Disease Report 2015" states that the number of Alzheimer's patients worldwide will increase from 46.8 million in 2015 to 131.5 million by 2050. It is predicted that the number of people will increase. Amid such predictions, people are paying much attention to maintaining and improving brain function.

Noradrenaline and serotonin are one of the neurotransmitters present in the central nervous system.
Regarding noradrenaline, it has been reported that the activation of noradrenaline receptors in rats and mice improves memory for situations and spaces (Non-Patent Document 1), and human studies have also shown that MHPG (a metabolite of noradrenaline) in the blood before and after a memory test. ) There is a report that a positive correlation was observed between changes in concentration and test results (Non-Patent Document 2), and it is known that noradrenaline has a very close relationship with brain functions centered on memory. It is also known that noradrenaline is synthesized in the locus coeruleus and is projected to many areas such as the cerebral cortex.
On the other hand, regarding serotonin, it has been reported that there is a positive correlation between the activity of serotonin receptors in the hippocampus and memory/learning (Non-patent Document 3), and serotonin and brain function in the hippocampus are also very closely related. It is known.

As a technique for improving brain function, for example, a technique using an oligopeptide mixture containing dipeptides and tripeptides as a food additive has been disclosed (Patent Document 1).

JP2015-208282A

Murchison CF et al., Cell, 2004 Apr. 2;117(1):131-43 Southwick SM et al., Am. J. Psychiatry, 2002 Aug;159(8):1420-2 Yifat Glikmann-Johnston et al., Front Pharmacol., 2015 Dec. 10;6:289 T. ichinose, et al., B.B.B., 2015, DOI : 10.1080/09168451.2015.1044932

Norepinephrine acts mainly on the cerebral cortex, and serotonin mainly acts on the hippocampus and is involved in brain function, so it is thought that if both act at the same time, they will have a better effect on brain function than if each acts individually.
Patent Document 1 is a technology that focuses on dopamine and noradrenaline, but this invention does not consider at all serotonin, which is considered to play a very important role in brain function.

As described above, conventional techniques have only considered catecholamines that are synthesized and metabolized in the body using tyrosine as a starting substance, and have not focused on serotonin, which is synthesized and metabolized using tryptophan as a starting substance. Therefore, the present invention aimed to provide a material that, when added to food, promotes the release of both noradrenaline and serotonin in the brain, thereby preventing cranial nerve diseases and improving brain function.

The inventor conducted extensive research focusing on dipeptides, which are said to have better absorption efficiency among peptides. As a result, certain dipeptides, namely Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln, Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr -Gly, His-Tyr, Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr-Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr promote the release of both noradrenaline and serotonin in the brain and are efficiently metabolized, thus completing the present invention.

That is, the present invention
(1) Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln, Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His-Tyr , Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr-Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr A food composition for preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain, which contains one or more dipeptides as active ingredients;
(2) The food composition according to (1), wherein the dipeptide is one or more selected from Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Tyr-Val, and Val-Tyr. ,
(3) The food composition according to (1), wherein the dipeptide is one or more selected from Tyr-Trp and Trp-Tyr;
(4) Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln, Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His-Tyr , Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr-Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr an agent for improving brain function by promoting the release of noradrenaline and serotonin in the brain, which contains one or more dipeptides as active ingredients;
(5) The brain function improvement described in (4), wherein the dipeptide is one or more selected from Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Tyr-Val, and Val-Tyr. agent,
(6) the brain function improving agent according to (4), wherein the dipeptide is one or more selected from Tyr-Trp and Trp-Tyr;
(7) Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln for use in preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain; Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His-Tyr, Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr- One or more dipeptides selected from Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr,
(8) The noradrenaline and serotonin according to (7), wherein the dipeptide is one or more selected from Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Tyr-Val, and Val-Tyr. A dipeptide for use in preventing cranial nerve diseases or improving brain function by promoting the release of
(9) The dipeptide is one or more selected from Tyr-Trp and Trp-Tyr, and for preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain as described in (7). dipeptide for use,
(10) Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln for use in preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain; Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His-Tyr, Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr- Use of one or more dipeptides selected from Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr;
It is.
In other words, the present invention
(11) Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln, Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His-Tyr , Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr-Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr A food composition for preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain, which contains one or more dipeptides as active ingredients;
(12) The food composition according to (11), wherein the dipeptide is one or more selected from Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Tyr-Val, and Val-Tyr. ,
(13) Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln, Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His-Tyr , Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr-Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr an agent for improving brain function by promoting the release of noradrenaline and serotonin in the brain, which contains one or more dipeptides as active ingredients;
(14) Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln for use in preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain; Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His-Tyr, Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr- One or more dipeptides selected from Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr and Leu-Tyr,
(15) The noradrenaline and serotonin according to (14), wherein the dipeptide is one or more selected from Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Tyr-Val, and Val-Tyr. A dipeptide for use in preventing cranial nerve diseases or improving brain function by promoting the release of
It is.

According to the present invention, Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln, Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly, His- Tyr, Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr-Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr, Leu-Tyr It has been found that both noradrenaline and serotonin are stimulated to be released in the brain and are efficiently metabolized, and the aforementioned peptides can be used in the form of pharmaceuticals or added to food or feed.

FIG. 3 is a diagram showing the results of measuring the concentration of 3-methoxy-4-hydroxyphenyl glycol (MHPG) in the cerebral cortex after oral administration of each dipeptide to mice. FIG. 2 is a diagram showing the results of measuring the concentration of 5-hydroxyindoleacetic acid (5-HIAA) in the cerebral cortex after oral administration of each dipeptide to mice.

(Food composition for preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain)
The food composition of the present invention for preventing cranial nerve diseases or improving brain function by promoting the release of noradrenaline and serotonin in the brain contains specific dipeptides as active ingredients, and these dipeptides, in addition to noradrenaline, release serotonin. It also promotes the release of into the brain.
Specifically, the dipeptides include Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Cys-Tyr, Tyr-Gln, Asn-Tyr, Glu-Tyr, Asp-Tyr, Cys-Tyr, Tyr-Gly. , His-Tyr, Thr-Tyr, Tyr-Thr, Tyr-Asp, Val-Tyr, Gly-Tyr, Tyr-Met, Tyr-Lys, Tyr-His, Tyr-Val, Arg-Tyr, Lys-Tyr, Leu -Tyr. Preferred are Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Tyr-Val, and Val-Tyr, more preferred are Tyr-Trp and Trp-Tyr. These dipeptides can be used alone or in combination of two or more.

Note that the dipeptide of the present invention may be expressed using the abbreviations of amino acids as shown below.
Tyr-Ala: YA, Ala-Tyr: YA, Arg-Tyr: RY, Asn-Tyr: NY, Tyr-Asp: YD, Asp-Tyr: DY, Tyr-Cys: YC, Cys-Tyr: CY, Tyr- Gln: YQ, Glu-Tyr: EY, Tyr-Gly: YG, Gly-Tyr: GY, Tyr-His: YH, His-Tyr: HY, Leu-Tyr: LY, Tyr-Lys: YK, Lys-Tyr: KY, Tyr-Met: YM, Tyr-Thr: YT, Thr-Tyr: TY, Tyr-Trp: YW, Trp-Tyr: WY, Tyr-Val: YV, Val-Tyr: VY.

Dipeptides can be synthesized from amino acids or obtained by hydrolyzing proteins.
Examples of means for hydrolyzing proteins include hydrolysis with protease, hydrolysis with acid, hydrolysis with alkali, etc. Hydrolysis with protease is preferred.
As raw materials for peptides, vegetable proteins and animal proteins can be used. For example, as raw materials for vegetable proteins, beans such as soybeans, peas, mung beans, and chickpeas, grains such as rice, corn, and wheat, and almonds are used as raw materials for peptides. , cashew nuts, walnuts, pistachios, hazelnuts, macadamia nuts, and other nuts, and raw materials for animal protein include beef, pork, chicken, eggs, milk, and the like.

(protease)
In the case of protein hydrolysis using protease, a slurry or aqueous solution of the above protein is used as a substrate and the protease treatment is performed. The protease used here is a protease classified into "metallic proteases", "acidic proteases", "thiol proteases", and "serine proteases", preferably "metallic proteases", regardless of its origin from animals, plants, or microorganisms. Proteases can be appropriately selected from among proteases classified as ``proteases,'' ``thiol proteases,'' and ``serine proteases.''

This classification of proteases is based on the type of amino acid in the active center, which is commonly used in the field of enzyme science. Representative examples of each include "metallic proteases" such as Bacillus neutral protease, Streptomyces neutral protease, Aspergillus neutral protease, and "samoase"; Examples of "protease" include bromelain, papain, etc., and examples of "serine protease" include trypsin, chymotrypsin, subtilisin, Streptomyces alkaline protease, "Alcalase", "Bioplase", etc. The classification of enzymes other than these can be confirmed based on the operating pH and reactivity with inhibitors. Enzymes with different active centers have significantly different sites of action on substrates, so it is possible to reduce "uncut residue" and efficiently obtain enzymatically decomposed products. Alternatively, by using enzymes from different origins (original organisms) in combination, enzymatic decomposition products can be produced more efficiently.

The reaction pH and reaction temperature of the protease treatment may be set according to the characteristics of the protease used, and the reaction pH may normally be carried out around the optimum pH and the reaction temperature may be carried out near the optimum temperature. The reaction can generally be carried out at a temperature of 20 to 80°C, preferably 40 to 60°C. After the reaction, the remaining enzyme activity is deactivated by heating at a temperature sufficient to deactivate the enzyme (approximately 60 to 170°C).

The reaction solution after protease treatment can be used as it is or after being concentrated, but it can usually be used in the form of a dry powder by sterilization, spray drying, freeze drying, etc. Sterilization is preferably carried out by heating, and the heating temperature is preferably 110 to 170°C, more preferably 130 to 170°C. The heating time is preferably 3 to 20 seconds. Further, the reaction solution can be adjusted to an arbitrary pH, and precipitates and suspended matter generated during pH adjustment can be removed by centrifugation, filtration, or the like. Further, it can also be purified using activated carbon or adsorption resin.

(hydrolysis with acid or alkali)
Conditions such as substrate concentration, enzyme amount, treatment temperature, pH, time, etc. in acid or alkali hydrolysis in protein hydrolysis can be set as appropriate.

(Prevention of cranial nerve diseases or improvement of brain function)
Cranial nerve diseases include higher brain dysfunctions such as memory disorders, attention disorders, executive function disorders, and social behavior disorders, as well as symptoms pathologically related to these disorders, such as cerebral infarction, head trauma, and cerebral infarction. These include vascular dementia, Alzheimer's disease, Parkinson's disease, schizophrenia, depression, and anxiety.
In addition, specific examples of brain function improvement effects include improved memory, improved learning ability, improved attention, stress tolerance, antidepressant effects, anti-anxiety effects, improved concentration, and improved sleep quality.

In the present invention, by using the above-mentioned specific dipeptide containing tyrosine, the concentration of intracerebral catecholamines in the cerebral cortex increases, specifically, the concentration of noradrenaline and serotonin in the cerebral cortex increases. By increasing the concentration of these catecholamines in the brain, the effect of preventing cranial nerve diseases or improving brain function can be expected.

The food composition of the present invention can be used in the form of a pharmaceutical or added to food or feed, mixed with other raw materials as appropriate, if necessary. When provided in the form of a pharmaceutical, it can be used in various forms such as liquid, powder, tablet, and capsule. When used as a mixture in food, it can be mixed with solid foods such as biscuits, cakes, breads, food bars, and meat products, or dissolved in water as a beverage, yogurt, etc. There is no problem when mixed with liquid or semi-solid foods such as pudding. It can also be used as a supplement by mixing carbohydrates, vitamins, minerals, etc. When provided in the form of a mixture with feed, it can be used by mixing with known feed, regardless of whether it is terrestrial or aquatic.

The present invention will be explained by describing examples below. Note that parts and percentages in the examples are based on weight unless otherwise specified.

The dipeptide used in the present invention was synthesized under contract by Genscript Japan Co., Ltd. The synthesized dipeptides are shown in Table 1.

(Table 1)

(Preparation of dipeptide aqueous solution)
Each dipeptide in Table 1 was prepared at 50 mM using ultrapure water (using an apparatus manufactured by Merck Millipore).

(How to raise mice)
An 8-week-old "C57BL/6J" was purchased from CLEA Japan Co., Ltd. Rearing was carried out for one week at 25±1°C under a 12-hour light/dark cycle (light period from 7:00 to 19:00, dark period from 19:00 to 7:00). The animals were given free access to food, and "CRF-1" (manufactured by Oriental Yeast Co., Ltd.) was fed as solid feed.

(Oral administration of each dipeptide aqueous solution to mice and removal of cerebral cortex)
According to the method described in Non-Patent Document 4, each dipeptide aqueous solution was orally administered to mice and the cerebral cortex was removed as follows. Control (ultrapure water) and each dipeptide aqueous solution were orally administered at a concentration of 1 g/ml. Thirty minutes after oral administration, the cerebral cortex was removed under anesthesia using an inhalation anesthetic solution "Isoflurane" (manufactured by Pfizer Inc.) and used for the following intracerebral catecholamine measurements. However, the cerebral cortex was stored at -80°C until sample processing.

(Measurement of catecholamines in the brain)
In accordance with the method described in Non-Patent Document 4, intracerebral catecholamines were measured by HPLC method as follows. 0.2 M perchloric acid (containing 0.1 mM EDTA·2Na) in an amount 5 times the weight of the cerebral cortex and 100 μg of isoprotenol were added to the cerebral cortex and thoroughly crushed. After crushing, the mixture was left on ice for 30 minutes and centrifuged at 20,000 x g for 30 minutes at 4°C. After centrifugation, 200 μl of the supernatant was collected, and after adding 35 μl of 0.2 M acetate buffer, it was filtered with a filter (0.22 μm, Merck Millipore) to remove proteins. After protein removal, the sample was subjected to "HPLC (HTEC-500)" (Acom Co., Ltd.) to extract MHPG (3-methoxy-4-hydroxyphenyl glycol), the final metabolite of noradrenaline, and 5, the final metabolite of serotonin. -HIAA (5-hydroxyindoleacetic acid) was measured to examine its effect on brain function.
The measurement results for MHPG are shown in Table 2 and FIG. 1, and the results for 5-HIAA are shown in Table 3 and FIG. 2.
Note that dipeptides in the tables and figures are expressed using amino acid abbreviations.

(Table 2) MHPG measurement results

As a result of measuring MHPG, which is the final metabolite of noradrenaline, Tyr-Ala (YA), Ala-Tyr (YA), Arg-Tyr (RY), Asn-Tyr(NY), Tyr-Asp(YD), Asp-Tyr(DY), Tyr-Cys(YC), Cys-Tyr(CY), Tyr-Gln(YQ), Gln-Tyr(QY), Tyr -Glu(YE), Glu-Tyr(EY), Tyr-Gly(YG), Gly-Tyr(GY), Tyr-His(YH), His-Tyr(HY), Tyr-Ile(YI), Ile- Tyr(IY), Leu-Tyr(LY), Tyr-Lys(YK), Lys-Tyr(KY), Tyr-Met(YM), Met-Tyr(MY), Tyr-Phe(YF), Phe-Tyr (FY), Tyr-Pro(YP), Pro-Tyr(PY), Tyr-Thr(YT), Thr-Tyr(TY), Tyr-Trp(YW), Trp-Tyr(WY), Tyr-Tyr( When aqueous solutions containing YY), Tyr-Val (YV), and Val-Tyr (VY) were administered, high values were shown (Table 2, Figure 1).

(Table 3) 5-HIAA measurement results

As a result of measuring 5-HIAA, which is the final metabolite of serotonin, Tyr-Ala (YA), Ala-Tyr (YA), Arg-Tyr ( RY), Asn-Tyr(NY), Tyr-Asp(YD), Asp-Tyr(DY), Tyr-Cys(YC), Cys-Tyr(CY), Tyr-Gln(YQ), Glu-Tyr(EY ), Tyr-Gly(YG), Gly-Tyr(GY), Tyr-His(YH), His-Tyr(HY), Leu-Tyr(LY), Tyr-Lys(YK), Lys-Tyr(KY) , Tyr-Met(YM), Tyr-Thr(YT), Thr-Tyr(TY), Tyr-Trp(YW), Trp-Tyr(WY), Tyr-Val(YV), Val-Tyr(VY) A high value was shown when the dissolved aqueous solution was administered (Table 3, Figure 2).

From the above two results, Tyr-Ala(YA), Ala-Tyr(YA), Arg-Tyr(RY), Asn-Tyr(NY), Tyr-Asp(YD), Asp-Tyr(DY), Tyr -Cys(YC), Cys-Tyr(CY), Tyr-Gln(YQ), Glu-Tyr(EY), Tyr-Gly(YG), Gly-Tyr(GY), Tyr-His(YH), His- Tyr(HY), Leu-Tyr(LY), Tyr-Lys(YK), Lys-Tyr(KY), Tyr-Met(YM), Tyr-Thr(YT), Thr-Tyr(TY), Tyr-Trp (YW), Trp-Tyr (WY), Tyr-Val (YV), and Val-Tyr (VY) were found to increase the intracortical concentrations of both MHPG and 5-HIAA.
In other words, these dipeptides were suggested to promote the release of both noradrenaline and serotonin in the brain, and to metabolize them more efficiently, thereby increasing the effect of improving brain function.
In addition, among the dipeptides, Tyr-Trp, Trp-Tyr, Tyr-Ala, Ala-Tyr, Tyr-Val, Val-Tyr are preferred because of their effectiveness in increasing the concentration of both MHPG and 5-HIAA in the cerebral cortex. It was found that Tyr-Trp and Trp-Tyr were better.

Claims (4)

The active ingredient is one or more dipeptides selected from Tyr-Trp, Trp-Tyr, Tyr-Ala, and Ala-Tyr, and promotes the release of noradrenaline and serotonin in the brain to prevent cranial nerve diseases or improve brain function. Food composition for improvement. The food composition according to claim 1, wherein the dipeptide is one or two selected from Tyr-Trp and Trp-Tyr. A brain function improving agent containing one or more dipeptides selected from Tyr-Trp, Trp-Tyr, Tyr-Ala and Ala-Tyr as an active ingredient and promoting the release of noradrenaline and serotonin in the brain. The brain function improving agent according to claim 3, wherein the dipeptide is one or two selected from Tyr-Trp and Trp-Tyr.

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