Composition for improving social defeat

Abstract

To provide a novel composition that can improve the conditions of individuals who have fallen into socially defeated situations.SOLUTION: A composition for improving socially defeated situations contains theanine. Preferably, the composition is for oral administration.SELECTED DRAWING: Figure 4

Description

Technical Field

[0001] The present invention relates to a composition for improving social defeat situations, particularly an oral administration composition containing theanine.

Background Art

[0002] In modern society, humans inevitably have interpersonal relationships with others (including family members, friends, seniors, colleagues, and related persons of other companies). At this time, if mental instability such as bullying (any form of harassment) or overwork continues, the period of being in the position of a mental weakling may continue for a long time. The social defeat situation is a situation pointed out in recent years. It means being placed in a state of mental defeat in complex interpersonal relationships, unable to take healthy thoughts and actions, and being driven into a sick state mentally and / or physically. In such a situation, one's mood drops mentally and reaches a physically sick state (such as hair loss). For example, when receiving social defeat stress similar to the social defeat situation, it is known that the intestinal flora changes as a physical effect (Non-Patent Document 1).

Prior Art Documents

Non-Patent Documents

[0003]

Non-Patent Document 1

Disclosure of the Invention

Problems to be Solved by the Invention

[0004] However, even if a social defeat situation occurs, it is not necessarily recognized as a disease, so it is not always medicated or appropriately treated. Therefore, the provision of an improvement composition that can be easily ingested is desired. This invention has been made in view of the above-mentioned problems, and its purpose is to provide a novel composition that can improve the condition of a person who has fallen into a state of social defeat. [Means for solving the problem]

[0005] The inventors diligently conducted research to solve the above problems and, as a result, discovered that theanine has the effect of improving social defeat situations, thus completing the present invention. The composition for improving social defeat according to the present invention is characterized by containing theanine. In this case, it is preferable that the theanine content is such that the effective intake amount is between 0.2 mg / kg body weight and 200 mg / kg body weight per day. Furthermore, it is preferable that the composition is for oral administration. [Effects of the Invention]

[0006] According to the present invention, a novel composition can be provided that can improve the condition of a person who has fallen into a state of social defeat. [Brief explanation of the drawing]

[0007] [Figure 1] This diagram shows the procedure for creating a mouse model of a socially defeated situation. [Figure 2] These are photographic diagrams showing the areas used in the SI test. (a) shows a 14cm x 24cm interaction zone and a 9cm x 9cm waiting zone, while (b) shows the partition tubes that prevent physical contact between B mice (mice that have experienced social defeat) and aggressor mice when they are confronted during the SI test, from the front (left side) and side (right side), respectively. [Figure 3]Representative results of examining the behavioral trajectories of mice in OF tests and SI tests are shown. (A) shows the behavioral trajectories of healthy mice that did not experience social defeat in the OF test (left), and in the SI test when the Anew mouse was absent (center) and when the Anew mouse was placed in the cage (right). (B) shows the behavioral trajectories of mice that experienced social defeat when they were given tap water in the OF test (left), and in the SI test when the Anew mouse was absent (center) and when the Anew mouse was placed in the cage (right). (C) shows the behavioral trajectories of mice that experienced social defeat when they were given theanine in the OF test (left), and in the SI test when the Anew mouse was absent (center) and when the Anew mouse was placed in the cage (right). [Figure 4] This graph shows the effect of theanine on the concentration of substances that significantly increase or decrease in the hair of socially defeated mice compared to the hair of control mice. (A) Corticosterone concentration, (B) Selenium disulfide concentration, and (C) Concentration of an unknown substance with (m / z) 144.866 are shown. [Modes for carrying out the invention]

[0008] Next, embodiments of the present invention will be described with reference to the figures and tables. The technical scope of the present invention is not limited by these embodiments, and it can be implemented in various forms without changing the gist of the invention. Theanine is a type of amino acid, a water-soluble white crystalline powder. Methods for obtaining theanine include extraction from tea leaves, organic synthesis (Chem. Pharm. Bull., 19(7), 1301-1307(1971)), treatment of a mixture of glutamine and ethylamine with glutaminase (Japanese Patent Publication No. 7-55154), culturing tea cell cultures in a medium containing ethylamine to increase theanine accumulation in the cell cultures while promoting cell proliferation (Japanese Patent Publication No. 5-123166), and replacing ethylamine with ethylamine derivatives such as ethylamine hydrochloride. Examples of tea leaves include green tea, oolong tea, and black tea. The theanine used in this invention can be obtained by any of the above methods.

[0009] L-, D-, and DL-forms of theanine can all be used. Of these, L-form theanine is preferred because it is approved as a food additive and is more economically feasible. Theanine can be used in any form, such as refined products, crude products, or extracts. The composition for improving social defeat according to the present invention can be provided as a pharmaceutical (including oral pharmaceuticals), quasi-drug, health food, functional food, supplement, confectionery for food and beverage consumption, etc. This composition may contain other active ingredients. Such active ingredients include, for example, vitamins such as vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin A, vitamin D, vitamin E, vitamin K, niacin, pantothenic acid, folic acid, biotin, and lycopene; stomachic agents such as betaine hydrochloride, carnitine chloride, and betanethol chloride; and herbal extracts or traditional Chinese medicines such as ginseng, coix seed, processed garlic, ephedra, nandina fruit, cinnamon bark, gentian, dried tangerine peel, swertia japonica, evodia, lanceolata, clove, aloe, hops, kakkonto, keishito, saikokeishito, maoto, shosaikoto, and shoseiryuto. Antipyretic analgesics such as aspirin, acetaminophen, ethenzamide, ibuprofen, and salicylamide; expectorants such as noscapine hydrochloride and noscapine; antidiarrheals such as acrinol, berberine chloride, creosote, tannic acid, scopolamine hydrobromide, methylatropine bromide, papaverine hydrochloride, and ethyl aminobenzoate; vasodilators such as dextromethorphan hydrobromide, tipepidine hibenzate, and dihydrocodeine phosphate; isotibenzyl hydrochloride, diphenylpyraline hydrochloride, diphenhydramine hydrochloride, chlorpheniramine maleate, and ali tartaric acid. Antihistamines such as memazine, cough suppressants such as dextromethorphan hydrobromide, tipepidine hibenzate, and dihydrocodeine phosphate, minerals such as calcium, sulfur, magnesium, zinc, selenium, and iron, proteins such as soy protein, egg white powder, and whey protein, amino acids such as glycine, alanine, arginine, aspartic acid, cystine, phenylalanine, taurine, tryptophan, theanine, and gamma-aminobutyric acid, fatty acids such as EPA, DHA, linoleic acid, gamma-linolenic acid, and alpha-linolenic acid, caramel color, gardenia color, and anthocyanin. Anine pigment, annatto pigment, paprika pigment, safflower pigment, red yeast rice pigment, flavonoid pigment, cochineal pigment, amaranth, erythrosine, allura red AC, new coccine, phloxine, rose bengal, acid red, tartrazine, sunset yellow FCF, fast green FCF, brilliant blue FCF, indigo carmine and other pigments, flavors such as various fruit flavors and essences, malic acid and its salts, tartaric acid and its salts, acetic acid and its salts, lactic acid and its salts, sodium chloride, glutamic acid and its salts, mirin, vinegar, natural fruit juice,Examples of active ingredients include, but are not limited to, seasonings such as chopped or powdered vegetables, fruits, and seafood; mushrooms such as Agaricus, Shiitake, Reishi, and Lion's Mane mushrooms or their extracts; preservatives; pH adjusters; and other functional materials such as chondroitin sulfate, glucosamine, ceramide, and hyaluronic acid. Active ingredients may be used individually or in combination of two or more. The amount of active ingredients is appropriately determined depending on the type of ingredient and its intended use.

[0010] Examples of raw materials that can be used in the present invention include carbohydrates, thickening polysaccharides, fruit juice, dried fruit, acidulants, flavorings, colorings, and gelling agents. As carbohydrates, granulated sugar, refined sugar, other disaccharides, sugar alcohols such as sorbitol, reduced starch syrup, acid-hydrolyzed starch syrup, enzyme-hydrolyzed starch syrup, monosaccharides, trisaccharides, oligosaccharides, trehalose, isomaltulose, etc. can be used. Examples of fruit juices include apple, grape, strawberry, kiwi, peach, and mandarin orange juices, as well as vegetable juices such as carrot, spinach, celery, bell pepper, kale, cabbage, and watercress, or mixtures thereof. As gelling agents, for example, gelatin and pectin can be used. As for gelatin, various sources such as animal bones and skins, and fish scales can be used. Other gelling agents can also be used as long as they achieve the objective of the present invention.

[0011] The present invention will be further explained below with reference to examples, but it is not limited to these. <Test Method and Test Results> 1. Creation of a mouse model representing a social defeat situation. Figure 1 shows an overview of the test method. We prepared 10 aggressor mice (mice that physically attack others (ICR mice): A mice) and 10 small mice that are more likely to be targeted for aggression (B mice, C57BL / 6 mice (B6 mice)), and assigned numbers from 1 to 10 to each individual (A1 to A10, B1 to B10). One A mouse and one B mouse were each housed in the same cage (D0). At this time, a partition board with a small transparent hole was installed in the middle of the housing cage. The reason for the partition board being transparent is to visually recognize the presence of the other party on the opposite side of the partition board at all times, and the reason for making a hole is to recognize it olfactory. Regarding the A mouse, it was housed in the same cage for 10 days between D1 and D10 below.

[0012] From the day before the A mouse attacked the B mouse (D0), breeding was started as described above. On the next day (D1), the B mouse was moved to the A mouse's side, and the A mouse was allowed to attack the B mouse for 5 to 10 minutes. After that, the B mouse was scolded on the opposite side of the partition board. 24 hours later (D2), the B mouse was sequentially moved to the cage of another A mouse, and the A mouse was allowed to attack the B mouse for 5 to 10 minutes. After that, the B mouse was scolded on the opposite side of the partition board. After repeating the attack on the B mouse by the A mouse for 10 days in this way (D1 - D10), a Social Interaction test (SI test) was conducted on the 11th day. The B mice were divided into two groups of 5 each. One group of B mice was given ion-exchanged water (control group (G1)), and the other group of B mice was given water in which theanine (5%) was dissolved (test group (G2)) by free drinking water.

[0013] 2. SI Test (1) Explanation of the test method Figure 2 shows the area diagram used in the SI test. In Figure 2(a), the upper Interaction zone indicates the contactable area, and the lower Comer zone indicates the waiting area. In the SI test, a new Anew mouse that was not used in the above "1. Social defeat situation model mouse" was prepared. As an aggressor mouse, CD-1 mice were newly screened. Using the screening criteria, the target CD-1 mice were screened to confirm the presence of aggressive characteristics. (i) The B mice that were attacked by the above-mentioned "1. Social defeat status model mice" were placed in the test room 1 hour before the start of the SI test. The test was conducted in a room isolated from external sound sources and always under red light. With a 30-second interval in between, the 150-second SI test was conducted twice. (ii) With no Anew mice present, a small cage was placed in the accessible area, and the B mice were placed at the "S" (start) position of the open field. A 150-second free exploration time (IT (A mouse -) during acclimation) was provided. (iii) The B mice were returned to their breeding cages, and within 30 seconds, the Anew mice were placed in the small cage and positioned in the accessible area. (iv) The B mice were placed in the Comer Zone and observed for 150 seconds (IT (A mouse +)). The movement trajectories of the B mice were recorded throughout the entire time. "Details of photography" will be described later. (v) Analysis was performed using behavioral analysis software (Matlab. <https: / / jp.mathworks.com / products / matlab.html>). From the movement trajectories of the B mice in (iv), the percentage of time spent in the 9 cm × 9 cm area in Figure 2 was plotted. It was defined that the higher the degree of social defeat, the longer the time spent in the area (9 cm × 9 cm) away from the A mice. (vi) After the test, the back hair was shaved with a clipper.

[0014] (2) Details of photography OF test: At the start of photography, the B mice were slowly placed into the open field (OF) with their heads along the wall. Acclimation was carried out for 10 seconds (photography ON). Photography was stopped, and the B mice were returned to their breeding cages. Next, the B mice were placed in a cylinder and slowly placed into the OF with their heads along the wall. Photography was conducted for 150 seconds (photography ON). Photography was stopped, and the B mice were returned to their breeding cages. Next, after placing the Anew mice, the B mice were slowly placed into the OF with their heads along the wall. Photography was conducted for 150 seconds (photography ON). After photography was stopped and the B mice were returned, the OF was cleaned with alcohol. The left side of Figures 3(A) to (C) shows representative results of the OF test. In both healthy mice and socially defeated mice (tap water intake and theanine intake), the OF test showed that each mouse moved within the entire movable area.

[0015] 3. SI Test Results The center and right sides of Figures 3(A) to (C) show representative results from the study of the behavioral trajectories of mouse B. In healthy mice that had not experienced social defeat (Figure 3(A)), they showed interest in other individuals (Anew mice), and therefore spent more time around the cage and had higher sociability scores when an unfamiliar mouse was inside the cage (Figure 3(A) right side) compared to when the cage was empty (Figure 3(A) center). In contrast, in mice that had experienced social defeat and been given tap water (Figure 3(B)), they moved further away from the cage and spent less time around the cage and more time in the corners of the box compared to when the cage was empty (Figure 3(B) center), resulting in lower sociability scores. Furthermore, mice that demonstrated social defeat and ingested theanine (Figure 3(C)) spent more time around the cage and had higher sociality scores when an unfamiliar mouse was in the cage (Figure 3(C) right side) compared to when the cage was empty (Figure 3(C) center). As described above, healthy mice were interested in Anew mice and often stayed near the accessible area (1.2 points), whereas tap water intake social defeat model mice initially showed interest in Anew mice, but the memory of being attacked was recalled, causing them to move away from the accessible area (0.03 points). Furthermore, mice that received free access to theanine scored significantly higher points and showed improved social defeat compared to mice that did not receive theanine, and were able to return to the control group. The SI test points for the control group (n=10) and the theanine ad libitum group (n=10) were 0.49±0.36 (mean ± standard deviation) and 0.88±0.24 (mean ± standard deviation), respectively. A significance test (two-group non-related t-test) showed a significant difference with P<0.05. The control group had a large standard error (SD) relative to the points (indicating individual differences), while the theanine ad libitum group had a small SD relative to the points, suggesting that social defeat was stably suppressed regardless of the individual.

[0016] 3. Measurement of components in body hair (1) Nanoparticle spraying The components of the body hair on the back, which was trimmed with clippers after the SI test, were measured. Nanoparticles were used as an ionization support agent (matrix) for the substances. The nanoparticles were prepared using a wet precipitation method to obtain monodisperse nanoparticles. For preparation, Macaulayite ultra-nanoparticles were prepared using FeCl2·4H2O and 3-aminopropyltriethoxysilane as materials. 2 g of FeCl2·4H2O was weighed and dissolved in 100 mL of distilled water to obtain a 10 mM aqueous solution. After thorough stirring at room temperature for approximately 10 minutes, it was mixed with 100 mL of 3-aminopropyltriethoxysilane (APTES) to complete the reaction. Subsequently, the solution was centrifuged for 60 minutes, the supernatant was discarded, and the precipitate was washed by repeatedly adding distilled water for redispersion, centrifuging, and discarding the supernatant several times. Next, the nanoparticles were suspended in methanol, and the supernatant was sprayed onto sample sections using an airbrush (GSI Creos). Since the spray was applied from a distance of 10 cm to 15 cm from the tissue, the methanol evaporated, and only the fine particle powder was sprayed onto the tissue.

[0017] (2) Imaging mass spectrometry Hair samples from C57BL / 6JJcl mice (8 weeks old; male) were used. The collected hair sections were placed on indium cinnamoxide (ITO) coated glass slides to prepare tissue sections. Imaging mass spectrometry measurements were performed using a mass spectrometer (tims-tof flex; Bruker Daltonics). The laser irradiation interval was set to 5 μm. The obtained signal was obtained from 100 laser shots. Figure 4 shows the results of a summary of signals with representative m / z values ​​for substances that increased or decreased when social defeat occurred compared to the control (Cont.) (G1), and returned to the control state upon administration of theanine (G2). Numerous strong signals were obtained from body hair sprayed with fine powder between (m / z) 100 and 800. In particular, the mass-to-charge ratio (m / z) of 347.174 shown in Figure 4(A) is that of corticosterone, a type of stress hormone. This signal was hardly detected in the control mouse (Cont.) and theanine-free drinking social defeat mouse, but was detected at a value more than 10 times higher in the social defeat mouse (G1). Theanine-free drinking social defeat mouse (G2) had a value about 3 times lower than the control. From this data, it was found that the social defeat state was improved in theanine-free drinking social defeat mouse. A similar situation to that of corticosterone was observed with selenium sulfide, shown in Figure 4(B). Furthermore, regarding the signal at (m / z) 144.866 shown in Figure 4(C), although the substance was not identified, it was detected in high concentrations in control mice (Cont.) and theanine-free drinking socially defeated mice (G2), while it was detected at lower concentrations in socially defeated mice (G1). This result also indicates that the socially defeated state improved in theanine-free drinking socially defeated mice.

[0018] As shown in Figures 4(A) and (B), substances that were significantly increased in the hair of socially defeated mice compared to control mice (corticosterone and selenium disulfide) were reduced to control mouse levels by theanine administration. Furthermore, as shown in Figure 4(C), a substance that was significantly decreased in the hair of socially defeated mice compared to control mice (an unknown substance with a m / z of 144.866) was increased to control mouse levels by theanine administration. These results indicate that mice in a socially defeated state were able to escape this state and return to a control state with theanine. Thus, according to this embodiment, we have been able to provide a novel composition (theanine) that can improve the condition of those who have fallen into a state of social defeat.

Claims

1. A composition for improving social defeat, characterized by containing theanine and suppressing the decrease in sociability scores and / or the increase in corticosterone production that occurs when experiencing social defeat.

2. The composition for improving a social defeat situation according to claim 1, wherein the theanine content is such that the effective intake amount is 0.2 mg / kg body weight to 200 mg / kg body weight per day.

3. The composition for improving a social defeat situation according to claim 2, wherein the composition is for oral administration.

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