Composition for improving mental state, method for producing composition for improving mental state, and use of eggplant fruit for improving mental state

Abstract

[Problem] The present invention addresses the problem of providing a novel composition having the effect of improving a mental state. [Solution] The present invention pertains to: a composition that is for improving a mental state, and that contains, as an active ingredient, a water-soluble component of the fruit of eggplant (Solanum melongena); and a production method that is for an eggplant-derived composition for improving a mental state, and that comprises squeezing an eggplant fruit to obtain a juice, or adding water to an eggplant fruit and squeezing the fruit to which water has been added, to obtain a juice.

Description

Technical Field

[0001] The present invention relates to a composition for improving mental state, a method for producing the composition for improving mental state, and use of eggplant fruits for improving mental state.

Background Art

[0002] According to a report released on World Health Day, cardiovascular diseases account for approximately 17 million deaths annually, which is equivalent to about one-third of the total deaths worldwide (Non-Patent Document 1). Hypertension accounts for 9.4 million deaths worldwide every year, with at least 45% due to heart disease and 51% due to stroke. Hypertension is also a contributing factor to kidney failure, premature death, and disability (Non-Patent Document 1). Therefore, prevention and management of hypertension are major public health issues worldwide. Some studies have shown that lifestyle modifications including dietary changes, exercise, and weight loss contribute to primary prevention of hypertension and significantly reduce blood pressure in people with hypertension (Non-Patent Documents 2-4). Functional foods with antihypertensive effects are also expected to prevent and manage hypertension (Non-Patent Document 5). In Japan, functional foods for people with blood pressure exceeding normal are commercially available as "Foods for Specified Health Use" approved by the government or "Foods with Nutritional Function Claims" indicated at the responsibility of food manufacturers themselves.

[0003] Eggplant (Solanum melongena) is a globally popular vegetable consumed daily and an agriculturally and economically important cultivated crop. Eggplant is low in energy, carbohydrates, and protein but rich in dietary fiber and minerals (Non-Patent Document 6), and is recommended for the purpose of reducing energy intake and body weight (BW: Body weight) to prevent type 2 diabetes (Non-Patent Documents 7-9). Clinical trials conducted in the past regarding the effectiveness of eggplant are two studies, one related to an investigation on the cholesterol-lowering effect in subjects with hypercholesterolemia (Non-Patent Document 10) and the other on the evaluation of effectiveness in reducing body fat in overweight women (Non-Patent Document 11).

[0004] Acetylcholine is well-known as a neurotransmitter and exists in almost all organisms, including not only mammals but also those used as food ingredients (Non-Patent Document 12). It has been reported that food materials such as eggplant, bamboo shoots (Phyllostachys bambusoides), and shiitake mushrooms (Lentinus eddoes) contain acetylcholine (Non-Patent Documents 12 and 13). The present inventors reported the isolation of acetylcholine and lactoylcholine from lactic acid-fermented foods having a blood pressure-lowering effect in spontaneously hypertensive rats (SHR), and reported the possibility that these choline esters are causative substances for the blood pressure-lowering effect (Non-Patent Documents 14 and 15). Also, recently, the blood pressure-lowering effect in SHR by oral administration of eggplant powder rich in acetylcholine was clarified, suggesting that acetylcholine in eggplant suppresses sympathetic nerve activity and causes a blood pressure-lowering effect (Non-Patent Document 16). Bethanechol, a cholinergic agonist, stimulates the parasympathetic nervous system via muscarinic acetylcholine receptors and lowers blood pressure (Non-Patent Document 17). Therefore, it was presumed that acetylcholine contained in orally administered eggplant enhances parasympathetic nerve activity, suppresses sympathetic nerve activity by an antagonistic action, and lowers blood pressure. Stress and psychological state (PS) are closely related to sympathetic nerve activity (Non-Patent Documents 18 and 19). Psychological factors are also related to blood pressure and may lead to hypertension (Non-Patent Document 20). Therefore, ingestion of eggplant may improve stress and psychological state in addition to blood pressure.

[0005] The present inventors have studied the antihypertensive and vasodilatory components contained in fermented buckwheat (lactic acid fermented product of buckwheat plants), and provided an extraction composition containing a quaternary alkylammonium compound mainly composed of a plurality of choline esters, including at least acetylcholine and propionylcholine. It has also been clarified that when purified acetylcholine, propionylcholine, butyrylcholine, and lactoylcholine are orally administered once to SHR, they exhibit an antihypertensive effect (Patent Documents 1 and 2). Furthermore, the present inventors have clarified that freeze-dried powders of edible plants such as eggplant and bamboo shoots, and extracts of edible plants with ethanol or hydrous ethanol contain active ingredients having an antihypertensive effect such as acetylcholine, and can be used for oral intake (Patent Document 3).

[0006] On the other hand, there are reports that a blood pressure increase suppressing composition (Patent Document 4) containing vegetable juice extracts such as tomato, carrot, spinach, and sprouted buckwheat juice as active ingredients, and a composition containing a water-soluble, sugar-free tomato extract can be used as an antihypertensive drug (Patent Document 5). However, all of these required a plurality of raw materials, needed to remove sugar from the extract, or were not easily preparable.

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Non-Patent Documents

[0008]

Non-Patent Document 1

非特許文献2

非特許文献3

非特許文献7

非特許文献8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 20

Summary of the Invention

Problems to be Solved by the Invention

[0009] The inventor has focused on the fact that if a composition that can be more easily prepared using fewer raw materials and has a more effective psychological state improvement effect can be obtained, it can be mass-produced at low cost as a health food or pharmaceutical such as a supplement, and has explored new materials having a psychological state improvement effect.

[0010] Therefore, an object of the present invention is to provide a new composition having a psychological state improvement effect.

Means for Solving the Problems

[0011] The main invention of the present invention for solving the above problems is a composition for improving psychological state, which contains a water-soluble component of the fruit of eggplant (Solanum melongena) as an active ingredient.

[0012] Regarding other problems disclosed in the present application and their solutions, they will be clarified by the embodiments of the invention and the drawings.

Effects of the Invention

[0013] According to the present invention, a new composition having a psychological state improvement effect can be provided.

Brief Description of the Drawings

[0014]

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Mode for Carrying Out the Invention

[0015] The contents of the embodiments of the present invention will be listed and described. The present invention has, for example, the following configurations. [Item 1] A composition for improving mental state, containing components derived from the fruit of Solanum melongena. [Item 2] The composition for improving mental state according to item 1, comprising a water-soluble component of the fruit of eggplant (Solanum melongena) as an active ingredient. [Item 3] The composition for improving mental state according to item 1, consisting of a water-soluble component of the fruit of eggplant (Solanum melongena). [Item 4] The composition for improving mental state according to any one of items 1 to 3, for oral ingestion in a dosage such that the amount of choline ester in the water-soluble component is 0.5 μg / kg body weight or more and 50 mg / kg body weight or less. [Item 5] The composition for improving mental state according to any one of items 1 to 4, which contains 5 μg or more and 50 mg or less of choline ester and is for oral ingestion. [Item 6] The composition for improving mental state according to item 4 or 5, wherein the choline ester is any one selected from acetylcholine, butyrylcholine, and propionylcholine. [Item 7] The composition for improving mental state according to any one of items 1 to 6, which is a dry powder. [Item 8] The composition for improving mental state according to any one of items 1 to 7, which is a food composition. [Item 9] A method for producing a composition for improving mental state containing an eggplant-derived component, comprising juicing the fruit of eggplant (Solanum melongena) to obtain a juice. [Item 10] A method for producing a composition for improving mental state containing an eggplant-derived component, comprising adding water to the fruit of eggplant (Solanum melongena) and juicing the fruit to which water has been added to obtain a juice. [Item 11] The method for producing a composition for improving mental state according to item 10, comprising crushing the fruit before or after adding the water to the fruit. [Item 12] The method for producing a composition for improving mental state according to any one of Items 9 to 11, which includes heating the fruit. [Item 13] The method for producing a composition for improving mental state according to any one of Items 9 to 12, which includes converting the squeezed juice into a dry powder. [Item 14] A composition for improving mental state produced by the production method according to any one of Items 9 to 13. [Item 15] Use of the fruit of Solanum melongena for improving mental state. [Item 16] The use according to Item 15 by oral ingestion of the fruit of the Solanum melongena. [Item 17] A composition for improving mental state containing, as an active ingredient, a water-soluble component of a food product using a high choline ester-containing agricultural product as a raw material.

[0016] Hereinafter, a composition for improving mental state and a method for producing the composition for improving mental state according to one embodiment will be described.

[0017] 1. Composition for improving mental state The composition for improving mental state according to one embodiment contains, as an active ingredient, a water-soluble component of the fruit of Solanum melongena. Further, the composition for improving mental state according to another embodiment may consist of the water-soluble component of the fruit of Solanum melongena.

[0018] Here, the water-soluble component of the fruit of Solanum melongena means the water-soluble component obtained by squeezing the fruit of Solanum melongena, but also means the water-soluble component contained in the squeezed juice obtained by adding water to the fruit of Solanum melongena and squeezing it. For example, even when the squeezed juice unavoidably contains water-insoluble components, or when a part of the water-soluble components unavoidably remains as residue during squeezing, the components contained in the squeezed juice correspond to the water-soluble components.

[0019] The eggplants used in this embodiment are not particularly limited. Generally, commonly consumed eggplants are preferred. For example, varieties such as Tosa Hawk, Shintaro, Ryoma, Izumisano Mizuna Nasu, Batten Nasu, Koryo Salad Eggplant (alias: Bishonen), Higo Murasaki, Oobancha Eggplant, Chikuyo, Senryo, etc. can be used.

[0020] In this embodiment, examples of choline esters that can be included in the composition for improving mental state include acetylcholine, butyrylcholine, propionylcholine, etc. Among these, a composition containing one or more of them may also be used.

[0021] In this embodiment, when the composition for improving mental state is used for humans, it is used for oral ingestion at a dose such that the amount of acetylcholine in the water-soluble component is 0.5 μg / kg body weight to 50 mg / kg body weight, preferably 2 μg / kg body weight to 20 mg / kg body weight, particularly preferably 5 μg / kg body weight to 1 mg / kg body weight, and more preferably 8 μg / kg body weight to 100 μg / kg body weight. By orally ingesting at such a dose, the effect of improving the mental state can be obtained more effectively.

[0022] In this embodiment, the composition for improving mental state is for oral ingestion and the choline ester content can be 5 μg or more and 50 mg or less. Also, in the composition for improving mental state, the choline ester content is preferably 500 μg or more and 20 mg or less, more preferably 1 μg or more and 10 mg or less. By orally ingesting at such a dose, the effect of improving the mental state can be obtained more effectively.

[0023] In this embodiment, in one aspect, the composition for improving mental state may be, for example, a dry powder such as a lyophilized powder or a hot air dried powder.

[0024] The composition for improving mental state of this embodiment can be used as a food composition such as various functional health foods or a pharmaceutical composition.

[0025] In the case of food, it may be used in combination with appropriate food additives. Moreover, not limited to such food compositions, it can be formulated into beverages by blending it with green tea, black tea, oolong tea, cereal tea, health drinks, sports drinks, etc., or into foods by blending it with biscuits, bread, candies, chocolates, macarons, etc., and provided in a form that can be ingested daily. Also, it can be used as a so-called supplement in an appropriate dosage form according to the preparation of the following pharmaceuticals.

[0026] When used as a pharmaceutical, it can be combined with appropriate pharmaceutical additives and used in various dosage forms according to the usual preparation methods. Examples of such dosage forms include oral dosage forms such as solid preparations like powders, granules, capsules, pills, tablets, etc., and liquid preparations like aqueous solutions, suspensions, emulsions, etc.

[0027] When the composition for improving mental state according to this embodiment is used as food, it can be used not only as general food and drink but also as foods for specified health use or foods with functional claims that exhibit specific functions and aim to promote health.

[0028] Specific forms in this case include supplements consisting of capsules, tablets, powders, granules, etc. containing the composition for improving mental state according to this embodiment as an active ingredient, bakery foods such as bread, cakes, cookies, macarons, etc., seasonings such as sauces, soups, dressings, mayonnaise, etc., dairy products such as milk, yogurt, creams, etc., confectioneries such as chocolates, candies, etc., or various beverages such as green tea, black tea, oolong tea, barley tea, cereal tea, fruit juice, vegetable drinks, milk drinks, soft drinks, and carbonated drinks.

[0029] The dosage when the composition for improving mental state according to this embodiment is used as an active ingredient of a pharmaceutical composition varies depending on the ratio of each component and also varies depending on various factors such as the patient's age, weight, gender, symptoms, administration method, etc. Also, it can be appropriately increased or decreased according to the degree of symptom improvement. The number of administrations can be from once a day to several times a day.

[0030] When the composition for improving mental state according to this embodiment is used as a food, the intake amount can be selected according to the case of oral administration of the above-mentioned pharmaceutical. However, in the case of food and drink, unlike pharmaceuticals, the dosage and the number of administrations are not particularly limited. Therefore, as long as particularly severe symptoms do not occur, the intake amount may be selected without limitation in consideration of the purpose of maintaining health and the taste and palatability.

[0031] The composition for improving mental state according to this embodiment can be used, for example, not only for the composition for improving mental state but also for subjects with hypertension and subjects who are healthy (not suffering from hypertension) but have high blood pressure.

[0032] Various additives can be added to the composition for improving mental state according to this embodiment, if necessary. As the additives, various ones can be used and are not particularly limited. For example, saccharides such as lactose hydrate, sucrose, glucose, reduced maltose, mannitol, sorbitol, starches such as corn starch, potato starch, partially alpha - modified starch, dextrin, pullulan and their derivatives, celluloses such as crystalline cellulose, microcrystalline cellulose, derivatives such as carboxymethyl cellulose, polyethylene glycol, and one or a mixture of two or more of magnesium aluminometasilicate can be mentioned. Among them, dextrin is more preferable. The additives may be used alone or in combination of two or more. These additives are preferably contained in an amount of 1 to 1000 parts by mass, more preferably 10 to 300 parts by mass, based on 100 parts by mass of the composition of the present invention.

[0033] Note that the cholinesterase, which is the active ingredient of the composition for improving mental state according to this embodiment, is not limited to the above-mentioned eggplants, and any of those derived from animals, plants, or microorganisms can be used. In particular, it is preferable to use those derived from organisms that humans have eaten, and it is preferable that they are derived from edible plants. That is, the composition for improving mental state according to this embodiment is not limited to being derived from eggplant fruits, and can include the water-soluble components of foods made from cholinesterase-rich agricultural products as the active ingredient.

[0034] As for edible plants other than eggplants, there is no particular limitation as long as they contain cholinesterase. Examples of edible plants include cucumber, tomato, bell pepper, green pepper, asparagus, sweet potato, cabbage, lettuce, carrot, apple, chili pepper, Japanese pear, grape, radish sprout, broccoli, alfalfa, mung bean sprout, buckwheat, bamboo shoot, etc. Among them, it is preferable that they are cholinesterase-rich agricultural products, and from the viewpoint of the acetylcholine content, bamboo shoot and Poaceae tribe bamboo are preferable, and it is preferable that the bamboo shoot is the tip of the spike.

[0035] 2. Manufacturing method of the composition for improving mental state In one aspect, the composition for improving mental state according to the above-mentioned present embodiment is obtained by the following manufacturing method.

[0036] The manufacturing method of the composition for improving mental state according to this embodiment includes squeezing the eggplant fruit to obtain a squeezed juice, or adding water to the eggplant fruit, and squeezing the fruit added with water. Even when manufacturing from raw materials other than eggplant fruits, the composition for improving mental state can be obtained by the same method.

[0037] The water added to the eggplant fruit is not particularly limited, and water within the temperature range of 5°C or higher and 40°C or lower may be added. It is preferable to add water at room temperature. The amount of water added is preferably 50 parts by mass or more and 200 parts by mass or less, more preferably 75 parts by mass or more and 150 parts by mass or less, and particularly preferably 100 parts by mass, based on 100 parts by mass of the raw eggplant fruit. If the amount of water is small, there is a risk that the extraction of the water-soluble components, which are the active ingredients, will be insufficient. If the amount of water is large, the processing volume will increase, and for example, it may become difficult to process into a dry powder.

[0038] The eggplant fruit is preferably crushed before or after adding water, and particularly preferably crushed before adding water from the viewpoint of ease of crushing. By crushing the eggplant fruit, the extraction of water-soluble components becomes easier. Crushing can be performed in various forms such as strip cutting, dice-shaped blocks, and paste form, but in order to make juicing easier, it is preferable to crush it into a paste form. For crushing, for example, a juicer, mixer, mill, crusher, etc. can be used.

[0039] The water used in the method for producing the composition for improving mental state according to this embodiment can be water generally used in food processing such as mineral water, distilled water, deionized water, ion-exchanged water, electrolyzed water, tap water, well water, and industrial water for food. The pH is not particularly limited, but in order to stably hold acetylcholine, a pH of 9.0 or lower and 3.0 or higher is preferable, preferably 8.0 or lower and 4.0 or higher, and more preferably 6.5 or lower and 4.5 or higher. Water whose pH is adjusted using a pH adjuster such as citric acid or ascorbic acid can be used.

[0040] Also, ethanol may be added to reduce bacterial contamination during the juicing operation. Increasing the ethanol concentration helps reduce microbial contamination, but since the solubility of the water extract deteriorates and the extraction efficiency of water-soluble components decreases, the ethanol concentration is preferably 9.9% or less, preferably 6% or less, and more preferably 4% or less.

[0041] In the method for producing a psychological state improving composition according to this embodiment, eggplant fruit may be heated. Heating inactivates acetylcholinesterase, allowing for efficient extraction of acetylcholine. Furthermore, the eggplant fruit tissue is softened, destroying the cell walls and facilitating extraction of water-soluble components and crushing the eggplant fruit.

[0042] The heating method is not particularly limited, and for example, heating can be carried out for 2 to 60 minutes at a temperature ranging from 30° C. to 120° C. The heating temperature is preferably from 80° C. to 100° C.

[0043] Examples of methods for heating eggplant fruit include heating in a microwave oven, on a hot plate or in a pan, immersing in boiling water, exposing eggplant to steam, etc. Among these, immersing in boiling water and exposing eggplant to steam are preferred.

[0044] Furthermore, the eggplant fruit may be heated before or after the addition of water, during heating with water, before crushing, after crushing, or during crushing. However, crushing the heated eggplant and then heating the crushed eggplant quickly is preferable for inactivating the acetylcholinesterase.

[0045] The method for extracting juice from crushed eggplant is not particularly limited as long as it can separate the juice from the residue. For example, the juice can be extracted using a strainer bag and a wine extractor, or by using a juice extractor. A centrifuge or screw press can also be used.

[0046] The obtained squeezed juice can be dried to produce a dried product in the form of a lump or powder. The drying method is not particularly limited, but it is preferable to dry the dried product until the moisture content is 0% by mass or more and 10% by mass or less. For example, drying can be performed by heat drying such as hot air drying, drum drying, freeze drying, spray drying, etc.

[0047] The dried product can be made into a dry powder by pulverization. The pulverization method is not particularly limited, but it is preferably possible to pulverize to about 5 to 100 mesh. For example, it can be pulverized using a mill.

[0048] The composition produced by the method for producing a composition for improving mental state according to the present embodiment tends to contain less dietary fiber and more carbohydrates. The ratio of dietary fiber to carbohydrates in the composition of the present invention is, for example, 1:2 to 1:100, preferably 1:5 to 1:40, and particularly preferably 1:10 to 1:20.

[0049] The composition for improving mental state containing eggplant-derived cholinesterase according to the present embodiment, when continuously ingested, significantly improves the blood pressure and mental state of normotensive hypertensives and grade I hypertensives who are feeling stressed. This is demonstrated, for example, by a placebo-controlled randomized double-blind parallel group comparison test as described in the examples below.

[0050] For example, regarding the improvement of blood pressure, by continuously ingesting the composition for improving mental state according to the present embodiment, for normotensive hypertensives, the blood pressure at the clinic (diastolic / post-load diastolic), home blood pressure (systolic / diastolic at waking / diastolic at bedtime) are significantly improved, and for grade I hypertensives, the blood pressure at the clinic (systolic / diastolic) is significantly improved. Overall, the blood pressure at the clinic (systolic / diastolic), home blood pressure (systolic / diastolic at waking) are significantly improved.

[0051] Regarding the improvement of mental state, by continuously ingesting the composition for improving mental state according to the present embodiment, for normotensive hypertensives, negative mental states such as "depression - feeling down", "confusion - bewilderment", "anger - hostility", "TMD score" are significantly improved, and for grade I hypertensives, positive mental states such as "friendliness", "vitality - vigor" are significantly improved. Overall, "confusion - bewilderment" is significantly improved.

[0052] Note that, as described later, there are no safety issues even if the composition for improving mental state according to the present embodiment is continuously ingested, and no side effects or harmful events that pose problems are observed during the 12-week ingestion period.

[0053] As described above, the composition for improving mental state according to the present embodiment and the method for producing the composition for improving mental state can provide a new composition having an effect of improving mental state.

[0054] Note that since the composition for improving mental state according to the above-described present embodiment is for oral ingestion, the effect of improving this mental state can also be obtained by eating the eggplant fruit itself. In this case, as described above, for example, by eating the eggplant fruit in a dose that can ingest 5 μg or more and 50 mg or less of choline ester content, an effect of improving mental state and other effects, for example, an effect of improving blood pressure can be obtained. The eggplant to be eaten can be eaten raw, heated, or can also be obtained as processed foods such as pickles.

[0055] 3. Examples Hereinafter, the present invention will be described more specifically by way of examples and comparative examples, but the present invention is not limited only to these examples. Note that "parts" and "%" in the examples and comparative examples are based on mass unless otherwise specified.

[0056] In this example, the purpose is to examine the long-term blood pressure, stress, and mental state improvement effects of eggplant in human subjects with prehypertension and / or stage I hypertension who are feeling stress through a randomized, double-blind, placebo-controlled, parallel group comparison study.

[0057] Blood pressure is classified according to the Hypertension Treatment Guidelines 2014 (edited by the Hypertension Treatment Guidelines Compilation Committee of the Japanese Society of Hypertension, April 2014, published by the Japanese Society of Hypertension, ISBN: 978-4-89775-322-5) and is defined as follows. Normal high blood pressure: 130 mmHg ≤ systolic blood pressure ≤ 139 mmHg and / or 85 mmHg ≤ diastolic blood pressure ≤ 89 mmHg; Grade I hypertension: 140 mmHg ≤ systolic blood pressure ≤ 159 mmHg and / or 90 mmHg ≤ diastolic blood pressure ≤ 99 mmHg

[0058] 3.1. Materials and Methods 3.1.1. Subjects A screening test was conducted on 189 people, and 100 Japanese men and women aged 35 or older and under 65, who had normal high blood pressure or Grade I hypertension and felt stressed, participated in this study. Subjects who met the following conditions were excluded from the study. · Those who are receiving instructions, treatment, or drug treatment from a doctor due to hypertension, schizophrenia, depressive disorder, manic disorder, arrhythmia, or bradycardia · Those with a BMI value of 30 kg / m 2 or higher · Premenopausal and postmenopausal women who complain of obvious physical changes · Those who may show allergic reactions to drugs or foods, especially those who may show allergic reactions to eggplant, secondary hypertension, severe cerebrovascular diseases, heart function disorders, liver disorders, kidney disorders, or gastrointestinal disorders · Those who are suffering from infectious diseases that require reporting to the authorities · Those with a history of major gastrointestinal surgery · Those with abnormal hypertension or hypotension, or abnormal blood data · Those with severe anemia · Those who regularly take medications, functional foods, or supplements due to blood pressure or stress · Those with alcoholism or eating disorders · Women who donated 400 mL of blood within 16 weeks · Men who donated 400 mL of blood within 12 weeks · Men and women who donated 200 mL of blood within 4 weeks · Those who are participating in another clinical trial or have participated in a clinical trial within 4 weeks · Pregnant or lactating women · Women who may become pregnant during this trial · Persons with any other diseases or health concerns that are the cause of problems in this test

[0059] Subjects who met the eligibility criteria were randomly assigned to the eggplant (capsules containing eggplant powder) group and the placebo (capsules containing dextrin) group by stratification of gender, age, systolic blood pressure, and diastolic blood pressure at the first visit. The attributes of the subjects in each group are shown in Table 1. The allocation of the test diet was performed by computerized blocking by a third-party data center (Media Education Center, Hokkaido Information Technology Research Institute, Esashi, Hokkaido). The clinical research collaborators were blinded to the allocation information during the period of this test. Key open was performed after the analysis of the data was completed.

[0060]

Table 1

[0061] 3.1.2. Power analysis of the number of cases Based on the effect size of 0.63 detected from the results of a preclinical trial conducted for 8 weeks (https: / / upload.umin.ac.jp / cgi-open-bin / ctr_e / ctr_view.cgi?recptno=R00003294 registration number UMIN000028785 registration date August 13, 2017), a sample size of 80 subjects (40 subjects per group) was calculated at a power of 80% and a significance level of 5%. Assuming that 20% of the subjects would drop out midway, the number of subjects was set at 100 (50 subjects per group).

[0062] 3.1.3. Preparation of eggplant dry powder capsules The test composition was prepared by the following method.

[0063] The eggplant powder was prepared from eggplant juice containing excipients for drying into a dry powder. The test food was defined by the daily intake of eggplant-derived choline ester (acetylcholine). The amount of acetylcholine to be ingested was set at 2.3 mg per day (equivalent to 22 g of fresh eggplant used in this test).

[0064] Eggplants as raw materials were sourced from Kochi Prefecture. Immediately after harvesting, the fresh eggplants were washed, shredded, and heated to prevent spoilage. They were then squeezed to remove solids, obtaining eggplant squeezed juice. After adding dextrin to the squeezed juice, it was freeze-dried. Each gram of the eggplant powder contained 1.92 mg of eggplant-derived choline ester (acetylcholine), and the yield was 4.08%. The eggplant powder was filled into capsules with a capacity of 300 mg. The placebo capsules were filled with only dextrin.

[0065] The capsules for each of the eggplant group and the placebo group were prepared according to a strictly quality-controlled protocol and had the same appearance. Also, the nutritional composition analysis of the capsules for each group was conducted at the Japan Food Analysis Center. Refer to Table 2 for the analysis results. Acetylcholine was analyzed using LC-MS / MS at Shinshu University. In this study, the subjects ingested 1200 mg of eggplant powder per day (300 mg capsules containing 25% dextrin-containing eggplant powder × 4 times). This is equivalent to the amount of eating approximately 22 g of fresh eggplants per day.

[0066]

Table 2

[0067] 3.1.4. Test This test was conducted at the Health Information Science Research Center of Hokkaido University of Information Sciences. The subjects ingested 4 tablets of eggplant powder capsules (2.3 mg as acetylcholine) or 4 tablets of placebo capsules every day for 12 weeks. The ingestion was 2 tablets each in the morning (after meal) and at night (before bedtime). At the baseline, 4th week, 8th week, 12th week, and 16th week (4 weeks after the final ingestion), physical examinations, vital signs and body composition measurements, urine tests, hematological and biological tests, and filling out questionnaires were carried out. Throughout the period of this test, the subjects were instructed to maintain their normal lives such as food intake and exercise habits, and not to ingest supplements, eggplants, bamboo shoots, etc. in the form of processed foods. Also, the subjects were asked to keep daily life records. Bamboo shoots are known to contain acetylcholine at a level similar to that of eggplants, and the subjects were not allowed to ingest foods with high acetylcholine content other than the test foods.

[0068] The main evaluation items are the blood pressure at the time of arrival, which consists of systolic blood pressure and diastolic blood pressure. The secondary evaluation items are home blood pressure (upon waking and at bedtime), stress evaluation using the Visual Analogue Scale (VAS), and psychological state evaluation using the full version of the Japanese version of the Profile of Mood States (POMS)-2 for adults.

[0069] 3.1.5. Quantification of blood pressure-lowering components in the Nasu capsules The quantification of acetylcholine was carried out by a modified method of the standard addition method described in International Publication No. 2018 / 070545. After adding (2-aminoethyl) trimethylammonium pivalamide (EN; synthesized at Shinshu University, internal standard) to the analysis sample, it was dissolved in 100 mM phosphate buffer and treated with a weak acidic cation exchange cartridge (Inertstep CBA 100 mg / 1 mL, manufactured by GL Sciences Inc., Tokyo). The fraction containing acetylcholine was eluted with 1 M hydrochloric acid and collected in a volumetric flask. When a certain amount was reached, it was divided into three parts, and different amounts of standard acetylcholine were added to create a dilution series.

[0070] Acetylcholine and EN contained in the sample were analyzed by LC-MS / MS under the following conditions. The separation was isocratic with the mobile phase (33% methanol containing 0.010% formic acid) using a YMC-Triart PFP column (4.6 mm × 250 mm, 5 μm, manufactured by WMC Co., Ltd., Kyoto). Acetylcholine and EN were detected using positive multiple reaction monitoring (MRM) with the following MRM transitions. 146.15>87.10 (acetylcholine) 187.30>128.15 (EN)

[0071] The quantified value of acetylcholine was corrected by the EN recovery rate. The quantification of γ-aminobutyric acid (GABA) and chlorogenic acid (CA) was detected in the same manner with the following MRM transitions. 104.15>87.20 (GABA) 355.10>163.20 (CA)

[0072] 3.1.6. Evaluation of Blood Pressure at the Time of Clinic Visit and Home Blood Pressure Blood pressure at the time of clinic visit was measured using an automated sphygmomanometer HEM-7080IC (manufactured by Omron Healthcare Co., Ltd., Kyoto, Japan) by a physician or a nurse after at least 10 minutes of rest, using the upper arm of the non-dominant arm. Three consecutive measurements were performed, and the median value of the measured values was obtained at each evaluation point.

[0073] The subjects measured their home blood pressure using an automated sphygmomanometer HEM-7080IC using the upper arm of the non-dominant arm. The subjects measured their blood pressure daily for 1 week prior to 2 to 6 visits, within 1 hour after waking up (morning blood pressure), and before going to bed (nighttime blood pressure). Three consecutive measurements were performed, and the median value of the measured values was obtained daily. The average blood pressure 3 days before each evaluation point was evaluated.

[0074] 3.1.7. Physical Examination, Blood Test, Biochemical Test, Urine Test Blood was collected from the subjects after 12 hours of fasting and used for blood tests of white blood cells (WBC), red blood cells (RBC), hemoglobin (Hb), hematocrit (Hct), and platelet count (Plt). Biochemical tests included liver function (aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (γ-GTP), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) levels). Renal function (blood urea nitrogen (BUN), creatinine (CRE), and uric acid (UA) levels), lipid profile (total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG) levels), blood glucose profile (fasting plasma glucose (FPG) and hemoglobin (Hb) Alc levels).

[0075] First morning urine was collected, and the sample was used for qualitative evaluation of pH, sugar, protein, occult blood, urobilinogen, and ketone.

[0076] Blood and urine tests were conducted at Sapporo Clinical Laboratory (Hokkaido, Japan). Body composition (BW, body fat rate (BFR), and BMI) was evaluated using a body composition analyzer DC-320 (Tanita, Tokyo, Japan).

[0077] 3.1.8. Mood profile assessment by POMS-2 and stress assessment by VAS questionnaire For the stress assessment of the subjects, a VAS questionnaire was used. The subjects were instructed to answer each question about their current physical and mental state by marking an "X" somewhere on a 100-mm line representing the worst state (left end, 0 mm) to the best state (right end, 100 mm). The results of the questionnaire were evaluated by the distance from the starting point of the left line to the "X", and it was evaluated that the greater the score number, the more the stress was improved.

[0078] The POMS-2 questionnaire was used for the purpose of evaluating the effect on the mental state by eggplant. The subjects were instructed to complete the questionnaire before consuming the test food. Anger-hostility, confusion-perplexity, depression-despondency, fatigue-apathy, and tension-anxiety were defined as negative mental states, while vigor-activity and friendliness were defined as positive mental states.

[0079] All mental states were evaluated by the full version of POMS-2 for adults (Success Bell Co., Ltd., Tokyo) consisting of 65 questions. The subjects answered about their activities in the past week from five options ranging from "not at all applicable" to "quite applicable". The total mood disturbance index (TMD) comprehensively representing negative mental states is the sum of the scores of anger-hostility, confusion-perplexity, depression-despondency, fatigue-apathy, and tension-anxiety, minus the scores of vigor-activity and friendliness.

[0080] To evaluate the anti-stress effect of eggplant, the subjects first took two capsules and then performed the Uchida-Kraepelin mental work test (UKT) for 15 minutes to induce mental stress. The VAS questionnaire was filled out twice, before consuming the test food and after performing the UKT.

[0081] 3.1.9. FFQg All subjects filled out the Food Frequency Questionnaire (FFQg) (Kensaku Co., Ltd., Tokyo, Japan) during the second to sixth visits. The FFQg was used to estimate the intake of nutrients (energy, protein, fat, carbohydrates, dietary fiber, salt) based on the subjects' usual diet. The subjects reported the amount and frequency of food consumed per week for approximately 29 food groups and 10 cooking methods.

[0082] 3.1.10. Ethics This clinical study was conducted in accordance with the Helsinki Declaration and the Ethical Guidelines for Clinical Research on Humans (Ministry of Education, Culture, Sports, Science and Technology). Prior to participating in this trial, the subjects carefully read the contents of the informed consent form for the test study and gave written informed consent. The research protocol was approved by the Ethics Committee of Hokkaido University of Information Sciences (Ebetsu City, Hokkaido) (approved on May 25, 2018. Approval number 2018-06). This study is already registered at https: / / upload.umin.ac.jp / cgi-open-bin / ctr_e / ctr_view.cgi?recptno=R000037995 (registration date July 10, registration number UMIN000033330).

[0083] 3.1.11. Statistical Analysis For the numerical analysis of the main evaluation items, secondary evaluation items, safety evaluation items, and dietary intake frequency questionnaires, the Student's t-test was used to compare the differences in subject data between the two groups. The differences in subject data were analyzed by repeating the analysis of variance method multiple times between groups. For subject attributes, the Fisher's exact probability test was used for gender, the Mann-Whitney U test was used for intake rates, and the Student's t-test was used for other attributes. Subgroup analysis was performed by dividing into subjects with systolic blood pressure of 130-139 mmHg or diastolic blood pressure of 85-89 mmHg (high-normal blood pressure subjects) and systolic blood pressure of 140-159 mmHg or diastolic blood pressure of 90-99 mmHg (grade I hypertension subjects) at the time of the visit. SPSS v25 (manufactured by IBM Japan, Ltd., Tokyo) was used for all statistical analyses. Exploratory data analysis using measured values was also performed in the same manner. All results were expressed as the sample mean ± standard error. A statistically significant difference was recognized when the p-value was 0.05 or less.

[0084] 3.2. Results 3.2.1. Subject Dropout and Characteristics Figure 2 shows the involvement of subjects and the research schedule during the test period. Volunteers who consented to informed consent (n = 189) were evaluated for eligibility, and 100 participants joined this study after screening. All subjects were randomly assigned to two groups (placebo group, n = 50; nas group, n = 50). Four subjects (personal reasons, n = 4) dropped out before the start of the test, and nine dropped out during the test period due to adverse events or personal reasons (in the placebo group, adverse events such as pneumonia (n = 1), colonic diverticulitis (n = 1), dyslipidemia (n = 1), urticaria (n = 1); in the nas group, adverse events such as injury (n = 2), mild liver dysfunction (n = 1), kidney disorder (n = 1) and personal reasons (n = 1)). Finally, a total of 87 subjects, 42 in the nas group and 45 in the placebo group, completed the test. Excluding the 4 subjects who dropped out due to personal reasons before the start of the test, 96 subjects were used for safety analysis. Ten subjects were excluded from the efficacy analysis. The reasons were as follows: outliers (outside the reference range (n = 1), more than 120% compared to the baseline of the safety evaluation items (n = 1)), mild diseases (allergies unrelated to the test food - (n = 2)), compliance problems or non - compliance with the criteria (n = 3), not feeling stressed at the start of the test (n = 1), lack of major evaluation item data due to unmeasured blood pressure (n = 2)). The efficacy analysis was performed on 36 subjects in the nas group and 41 subjects in the placebo group. The sex ratio, mean age, height, BMI, systolic blood pressure at the time of visit, diastolic blood pressure at the time of visit, and intake rate of each group are shown in Table 1. No significant differences were observed between the two groups for these indicators.

[0085] 3.2.2. Efficacy of Nas in Blood Pressure at the Time of Visit Figures 3 - 7 show the change amount from week 0 of blood pressure at the time of visit (Figures 3, 4) and the measured values (Figures 5 - 7). Figure 3 shows the diastolic blood pressure of all subjects, Figure 4 shows the diastolic blood pressure of subjects with high - normal blood pressure, Figure 5 shows the systolic blood pressure of all subjects, Figure 6 shows the systolic blood pressure of grade I hypertensive patients, and Figure 7 shows the diastolic blood pressure of grade I hypertensive patients. The values are shown as mean ± standard error. Statistical significance by Student's t - test: * p < 0.05 and *** p < 0.001 vs placebo group.

[0086] At the 8th week of eggplant powder intake, the diastolic blood pressure of all subjects was significantly improved compared with the placebo group (p = 0.024) (Figure 3, Table S1; Figures 18, 19). In the subgroup analysis, at the 8th week of eggplant powder intake, the increase in diastolic blood pressure in subjects with high-normal blood pressure was significantly suppressed (placebo: n = 26; eggplant: n = 27) (Figure 4, Table S1; Figures 18, 19). In the exploratory data analysis, at the 12th week of intake, the eggplant group showed significantly lower systolic blood pressure than the placebo group (p = 0.046) (Figure 5, Table S2; Figures 20 - 22). The systolic and diastolic blood pressures of subjects with grade I hypertension (placebo: n = 15; eggplant: n = 9) were significantly lower in the eggplant group than in the placebo group at the 12th week of intake (systolic blood pressure: p = 0.037, diastolic blood pressure: p = 0.041) (Figures 6, 7, Table S2; Figures 20 - 22). Furthermore, significant time - food interactions in the systolic blood pressure at the time of visit (change amount and measured value, p = 0.018), in subjects with grade I hypertension (change amount and measured value, p = 0.043), and in the diastolic blood pressure at the time of visit in subjects with high-normal blood pressure (change amount and measured value, p = 0.028) also supported the blood pressure lowering effect of eggplant powder intake (Table S1; Figures 18, 19 and Table S2; Figures 20 - 22).

[0087] 3.2.3. Efficacy of Eggplant in Home Blood Pressure Figures 8 - 10 show the change amount of home blood pressure from week 0. Figure 8 shows the systolic and diastolic blood pressures at waking in all subjects, Figure 9 shows the systolic and diastolic blood pressures at waking in subjects with high-normal blood pressure, and Figure 10 shows the diastolic blood pressure at bedtime in subjects with high-normal blood pressure. Values are shown as mean ± standard error. Statistical significance by Student's t-test: * p < 0.05 and *** p < 0.01 vs placebo group.

[0088] At the 4th week of eggplant powder intake, the systolic blood pressure (Figure 8, p = 0.017) and diastolic blood pressure (Figure 8, p = 0.032) of all subjects upon waking up were significantly decreased compared with the placebo group. In the subgroup analysis, the subjects with high-normal blood pressure in the eggplant group had significantly decreased systolic blood pressure (Figure 9, p = 0.041) and diastolic blood pressure (Figure 9, p = 0.008) upon waking up at the 4th week, and the diastolic blood pressure at bedtime was significantly suppressed at the 4th and 8th weeks (Figure 10, p = 0.029), compared with the placebo group. There was no significant difference in the measured values between the placebo group and the eggplant group.

[0089] 3.2.4. Efficacy of eggplant on stress and blood pressure The effects of eggplant on stress and mental state were evaluated by the VAS test indicating the degree of stress and the POMS-2 test indicating the degree of mental state (Table S3; Figures 23 - 26, Table S4; Figures 27 - 29). No difference in VAS scores was observed between the two groups at any time before and after the implementation of UKT. In the subgroup analysis of the POMS-2 test results, in the subjects with grade I hypertension in the eggplant group, "vitality - vigor" at the 4th week and "friendliness" at the 8th week were significantly better than those in the placebo group. In the exploratory data analysis, "confusion - bewilderment" was significantly improved in all subjects at the 12th week, and the subjects with high-normal blood pressure in the eggplant group had significantly improved negative mental states ("confusion - bewilderment", "depression - despondency" at the 8th week, "anger - hostility", "TMD" at the 12th week) (Figures 11 - 17).

[0090] 3.2.5. Nutritional evaluation of subjects during the study The nutritional status of the subjects was evaluated by FFQg (Table S5; Figure 30). The lipids and dietary fiber in the eggplant group at the 12th week were significantly lower than those in the placebo group (p = 0.009 and 0.035). However, there was no statistical significant difference in the intakes of energy, protein, carbohydrates, and salt between the eggplant group and the placebo group.

[0091] 3.2.6. Safety evaluation of eggplant To analyze the safety of eggplant powder intake, the body composition, complete blood count, liver function, kidney function, lipid profile, and blood glucose profile of the subjects were evaluated (Table S6; Figures 31, 32). Additionally, urine parameters were qualitatively evaluated (Table S6; Figures 31, 32). All fluctuations in values were within the range of reference values. Although adverse events were observed in each group, it was inferred based on the protocol criteria that none of the adverse events were related to the test food. Therefore, the intake of eggplant powder equivalent to 22 g of fresh eggplant per day and containing 2.3 mg of eggplant-derived choline ester (acetylcholine) was either free of adverse events or had minimal adverse events.

[0092] 3.3. Discussion A placebo-controlled double-blind parallel-group comparison trial revealed that continuous intake of eggplant powder containing 2.3 mg of eggplant-derived choline ester (acetylcholine) per day improved the blood pressure of subjects with high-normal blood pressure and grade I hypertension. Subgroup analysis confirmed the blood pressure improvement effect of eggplant powder intake in subjects with high-normal blood pressure. Additionally, based on a thorough literature search, it was confirmed that this study is the first clinical trial to show evidence of blood pressure improvement by eggplant. Furthermore, this study showed that eggplant intake improves both positive and negative psychological states.

[0093] In this study, the diastolic blood pressure at arrival, which is the primary evaluation item, was improved in all subjects by eggplant powder intake, and the effect was particularly high in subjects with high-normal blood pressure. Interestingly, the improvement effect of diastolic blood pressure by eggplant intake was more prominent in subjects with high-normal blood pressure (p < 0.001) than in all subjects (p = 0.024). The systolic blood pressure at arrival was improved in all subjects and in subjects with grade I hypertension, and the diastolic blood pressure at arrival in subjects with grade I hypertension was also improved. These were revealed by exploratory data analysis. The fact that eggplant intake improved the diastolic blood pressure in subjects with high-normal blood pressure and the systolic and diastolic blood pressures in subjects with grade I hypertension implies that eggplant may have different blood pressure improvement effects on different blood pressure classifications.

[0094] Improvements were also observed in the secondary evaluation items of home systolic blood pressure and diastolic blood pressure after eggplant intake. Improvements were seen in the systolic and diastolic blood pressures upon waking of all subjects and subjects with high-normal blood pressure, and in the diastolic blood pressure at bedtime of subjects with high-normal blood pressure, resulting in findings that support the effect of eggplant intake on improving blood pressure at arrival.

[0095] Management of early morning blood pressure is important for the prevention of heart failure [White, W.B. Clinical assessment of early morning blood pressure in patients with hypertension. Prev. Cardiol. 2007, 10, 210-214.], [Wang, J.G.; Kario, K.; Chen, C.H.; Park, J.B.; Hoshide, S.; Huo, Y.; Lee, H.Y.; Li, Y.; Mogi, M.; Munakata, M.; et al. Management of morning hypertension: A consensus statement of an Asian expert panel. J. Clin. Hypertens. 2018, 20, 39-44.].There are reports that the early morning blood pressure and nocturnal blood pressure of patients with anxiety disorder are higher than those of healthy people [Kayano, H.; Koba, S.; Matsui, T.; Fukuoka, H.; Toshida, T.; Sakai, T.; Akutsu, Y.; Tanno, K.; Geshi, E.; Kobayashi, Y. Anxiety disorder is associated with nocturnal and early morning hypertension with or without morning surge: Ambulatory blood pressure monitoring. Circ. J. 2012, 76, 1670-1677.], [Mellman, T.A.; Brown, D.D.; Jenifer, E.S.; Hipolito, M.M.S.; Randall, O.S. Posttraumatic stress disorder and nocturnal blood pressure dipping in young adult African-Americans. Psychosom. Med. 2019, 71, 627-630.]. As one of the contributing factors, sleep disorders caused by sympathetic nerve hyperactivity [Veith, R.C.; Lewis, N.; Linares, O.A.; Barnes, R.F.; Raskind, M.A.; Villacres, E.C.; Murburg, M.M.; Ashleigh, E.A.; Castillo, S.; Peskind, E.R.; et al. Sympathetic nervous system activity in major depression: Basal and desipramine-induced alterations in plasma norepinephrine kinetics. Arch. Gen. Psychiatry. 1994, 51, 411-422.] are assumed. Therefore, it was speculated that the intake of eggplant suppressed the sympathetic nerve activity during sleep of the subjects, thereby improving the early morning blood pressure of the stressed subjects.

[0096] Previously, the present inventors reported the antihypertensive effect of eggplant on SHR. By orally administering a low dose of eggplant powder (0.0821 mg / kg BW) daily, the increase in systolic and diastolic blood pressures of SHR was significantly suppressed, and according to Kleiber's law [Kleiber, M. The Fire of Life: An Introduction to Animal Energetics; Wiley: New York, NY, USA, 1961.], the effective daily dose for an adult (60 kg) was calculated to be 13.1 mg when converted to humans. The effectiveness of acetylcholine in eggplant may be lower for human individuals with high-normal blood pressure or stage I hypertension compared to SHR with moderate to severe hypertension. In this study using SHR, acetylcholine in eggplant acted on the M3 muscarinic acetylcholine receptor (M3 mAChR) and decreased the amount of urinary catecholamine. This indicates that acetylcholine in eggplant suppressed sympathetic nerve activity via M3 mAChR in the digestive system, resulting in a decrease in blood pressure. M3 mAChR is present in the epithelial cells of the stomach and intestines [Wess, J. Muscarinic acetylcholine receptor knockout mice: Novel phenotypes and clinical implications. Ann. Rev. Pharmacol. Toxicol. 2004, 44, 423-450.]. When orally administered acetylcholine derived from eggplant acts on M3 mAChR and stimulates the parasympathetic nervous system, when this stimulation reaches the medulla oblongata, sympathetic nerve activity is automatically decreased by reciprocal innervation.

[0097] Disorders of autonomic regulation are thought to contribute to the elevated blood pressure in SHR. There are reports that the amount of catecholamines in the blood of SHR is higher than that in normotensive Wistar-Kyoto (WKY) rats, which is due to the abnormal overactivity of the sympathetic nerves in SHR [Tsunoda, M.; Takezawa, K.; Ina, Y.; Nagashima, K.; Ohomori, K.; Kobayashi, S.; Imai, K. New approach for measurement of sympathetic nervous abnormality in conscious, spontaneously hypertensive rats. Jpn. J. Pharmacol. 2000, 83, 39-45.], [Donohue, S.J.; Stitzel, R.E.; Head, R.J. Time course of changes in the norepinephrine content of tissues from spontaneously hypertensive and Wistar Kyoto rats. J. Pharmacol. Exp. Ther. 1988, 245, 24-31.]. There are also reports that the sensitivity of the baroreceptor reflex in SHR is lower than that in WKY rats [Kumagai, H.; Averill, D.B.; Ferrario, C.M. Renal nerve activity in rats with spontaneous hypertension: Effect of converting enzyme inhibitor. Am. J. Physiol. 1992, 263, 109-115.]. This suggests that the neural regulation of blood pressure control varies depending on the level of hypertension, and that acetylcholine in the nasopharynx reduces blood pressure more effectively in patients with stage I hypertension than in those with high-normal blood pressure. Regarding the office blood pressure, which is the main evaluation item, nasal intake improved only the diastolic blood pressure in subjects with high-normal blood pressure, but improved both systolic and diastolic blood pressures in subjects with stage I hypertension.The differences in blood pressure between the eggplant group and the placebo group were 5.6 mmHg in diastolic blood pressure of subjects with high-normal blood pressure, 7.6 mmHg in systolic blood pressure and 7.3 mmHg in diastolic blood pressure of subjects with grade I hypertension.

[0098] Stress and negative mental states enhance sympathetic nerve activity. Therefore, it is thought that suppression of sympathetic nerve activity leads to improvement of stress and negative mental states. From this, in this study, subjects experiencing stress were recruited, and the improvement effects of stress and mental states in the nose were evaluated using the VAS and POMS-2 tests. The very simple VAS test has been used in other studies for the purpose of evaluating stress after oral intake of drugs [Diaper, A.; Papadopoulos, A.; Rich, A.S.; Dawson, G.R.; Dourish, C.T.; Nutt, D.J.; Bailey, J.E. The effect of a clinically effective and non-effective dose of lorazepam on 7.5% CO2-induced anxiety. Hum. Psychopharmacol. 2012, 27, 540-548.].Since mental state is involved in the onset and condition of hypertension [Andrew, M.J.; Baker, R.A.; Kneebone, A.C.; Knight, J.L. Mood state as a predictor of neuropsychological deficits following cardiac surgery. J. Psychosom. Res. 2000, 48, 537-546.], [Hassan, K.; Elimeleh, Y.; Shehadeh, M.; Fadi, H.; Rubinchik, I. The relationship between hydration status, male sexual dysfunction and depression in hemodialysis patients. Ther. Clin. Risk Manag. 2018, 14, 523-529.], the POMS-2 test was used to evaluate mental state [Yu, B.H.; Dimsdale, J.E.; Mills, P.J. Psychological states and lymphocyte beta-adrenergic receptor responsiveness. Neuropsychopharmacology 1999, 21, 147-152.].

[0099] The results of the VAS and POMS-2 tests showed that long-term intake of eggplant powder has no effect on stress but has the effect of improving the mental state. The anti-stress effect of eggplant should be examined by objective methods rather than by subjective evaluation in the VAS test. In the POMS-2 test, eggplant powder intake was improved in negative mental states ("confusion - bewilderment" of all subjects, "depression - despondency", "confusion - bewilderment", "anger - hostility", "TMD" of subjects with high-normal blood pressure) and positive mental states ("friendliness" and "vitality - vigor" of subjects with grade I hypertension). Also, the results of POMS-2 showed that the effectiveness of eggplant differed for subjects in each blood pressure classification. The tendency of mental state improvement from week 0 to week 4 suggests the adaptation effect of subjects to the test. At week 8, a deterioration of the mental state was observed. Although it had no effect on the whole test, this may be the effect of the earthquake that occurred 15 days before week 8 of the test (September 6).

[0100] In the dietary intervention of taking eggplant powder capsules in this study, the intake of foods rich in acetylcholine such as eggplant and bamboo shoots was prohibited. The capsules taken daily by the subjects in the eggplant group contained 1.2 g of eggplant powder, with an energy of 4.39 kcal, and the energy of the placebo capsules without eggplant powder was 4.35 kcal. Since there was no difference in nutritional value between the two, it was concluded that the improvement effect on blood pressure and psychological state was due to the eggplant powder. Eggplant contains not only acetylcholine but also GABA [Horie, H.; Ando, S.; Saito, T. The contents of γ-amino butyric acid in eggplant and its accumulation with heat treatment. J. Jpn. Soc. Food Sci. 2013, 60, 661-664.] and CA [Singh, A.P.; Luthria, D.; Wilson, T.; Vorsa, N.; Singh, V.; Banuelos, G.S.; Pasakdee, S. Polyphenols content and antioxidant capacity of eggplant pulp. Food Chem. 2009, 114, 955-961.] with antihypertensive effects. According to the Japanese Functional Food Labeling System, the effective doses showing the antihypertensive effects of GABA and coffee-derived CA in adults are 12.3 mg and 271 mg per day, respectively. The amounts of GABA and CA ingested from the eggplant powder used in this study were 7.65 mg (62.2% of the effective dose) and 12.3 mg (4.54% of the effective dose) per day, respectively, and in both cases, the amounts were less than the effective doses. The inventors have reported that the main compounds causing the antihypertensive effect of eggplant in SHR are not GABA and CA, but eggplant-derived choline ester (acetylcholine) (Non-Patent Document 16).

[0101] This result should be applicable when considering the blood pressure-lowering components of eggplant in this clinical trial using subjects with elevated blood pressure. Since the amounts of GABA and CA contained in the eggplant powder used in this clinical trial are small, neither of them alone can be expected to have a blood pressure-lowering effect. Also, there is no report of a synergistic effect that enhances the blood pressure-lowering effect when both coexist. Therefore, it is reasonable to assume that the blood pressure-lowering effect is due to acetylcholine. Therefore, the blood pressure-improving effects of GABA and CA in the eggplant powder used in this clinical trial are limited, and it was concluded that the main antihypertensive component of the eggplant powder is acetylcholine. Research on the antihypertensive effect of orally administered acetylcholine is currently underway, and positive results are expected. In this randomized comparative trial, the safety of continuous intake of eggplant powder equivalent to 22 g of fresh eggplant per day was also confirmed.

[0102] In conclusion, continuous intake of eggplant powder significantly improves blood pressure and mental state. The clinic and home blood pressures of all subjects and subjects with high-normal blood pressure were improved at the 4th and 8th weeks. This transient improvement is thought to be due to environmental factors. It is a well-known fact that clinic and home blood pressures vary with seasons [Lewington, S.; Li, L.; Sherliker, P.; Guo, Y.; Millwood, L.; Bian, Z.; Whitlock, G.; Yang, L.; Collins, R.; Chen, J.; et al. Seasonal variation in blood pressure and its relationship with outdoor temperature in 10 diverse regions of China: The China Kadoorie Biobank. J. Hypertens. 2012, 30, 1383-1391.]. Blood pressure rises as the temperature drops from summer to winter. This trial was conducted from summer to the beginning of winter, and the average temperature decreased from 27.9°C to 9.6°C. This is considered to correspond to a 3.5 mmHg increase in blood pressure.

[0103] The increase in blood pressure due to a decrease in temperature and the decrease in blood pressure due to the intake of eggplant powder were considered to be caused through the regulation of the nervous system and were in a competitive relationship with each other. Therefore, it was considered that the effect of eggplant after the 4th or 8th week from the start of the test was attenuated. The blood pressure at the time of visit of all subjects and subjects with grade I hypertension improved at the 12th week. This should be considered the effect of continuous intake of eggplant powder. The effect of eggplant powder is mild and gradually takes effect, which is distinct from the effect of antihypertensive drugs. Therefore, continuous intake of eggplant is expected to improve the blood pressure of people with high-normal blood pressure or grade I hypertension. Eggplant is regarded as a safe food. In this test as well, the safety was confirmed as well as the improvement effect of eggplant powder on blood pressure and mental state. These new effects support the usefulness of eggplant powder rich in acetylcholine as a safe approach to blood pressure and mental health.

[0104] 3.4. Summary By means of a placebo-controlled double-blind parallel-group comparison test, it was confirmed that continuous intake of eggplant powder for 12 weeks improved blood pressure and mental state without side effects. The eggplant-derived choline ester (acetylcholine) contained in 1.2 g of eggplant powder ingested per day at 2.3 mg can be said to be a new functional food factor. Also, this study became the first study to prove the antihypertensive effect and the effect of improving mental state of eggplant. In the future, it is expected that eggplant will be used as a safe approach to blood pressure and mental health.

[0105] As described above, the composition according to this embodiment can provide a new composition having an excellent antihypertensive effect and an effect of improving mental state, which can be easily and inexpensively produced from the fruit of eggplant (Solanum melongena). Also, in oral administration, it has a more remarkable antihypertensive effect than a composition having an antihypertensive effect with a conventional choline ester as an active ingredient, and a functional food with claims or a therapeutic drug for hypertension and the like can be produced using this.

[0106] The eggplant fruit, which is a raw material of the composition according to this embodiment, is widely consumed as food. The composition of the present invention can be composed only of components derived from eggplant, has high safety, and can be continuously ingested over a long period. Further, the composition of the present invention can be used in foods, drinks, and pharmaceuticals in various forms, such as an aqueous extract of eggplant fruit, a concentrated extract obtained by concentrating the extract, a dry powder (freeze-dried powder, hot air-dried powder, spray-dried powder, etc.), and a suspension of the dry powder.

Claims

Claim 1 A composition for improving mental state, comprising a water-soluble component of the fruit of Solanum melongena as an active ingredient, wherein the water-soluble component contains choline ester, and administration of the composition improves a negative mental state. Claim 2. The composition according to claim 1, wherein the negative mental state is at least any one of confusion - bewilderment, depression - dejection, and anger - hostility in the Profile of Mood States (POMS-2) test. Claim 3 The composition for improving mental state according to claim 1 or 2, for oral ingestion in a dosage such that the amount of choline ester in the water-soluble component is 0.5 μg / kg body weight or more and 50 mg / kg body weight or less. Claim 4. The composition for improving mental state according to claim 1 or 2, wherein the choline ester is any one selected from acetylcholine, butyrylcholine, and propionylcholine, and the choline ester content is 5 μg or more and 50 mg or less. Claim 5 The composition for improving mental state according to claim 1 or 2, which is a dry powder. Claim 6 The composition for improving mental state according to claim 1 or 2, which is a food composition.

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