Composition and use thereof

Abstract

A composition comprising a diacyl glyceryl ether (DAGE) represented by the Formula I, where R1 is an aliphatic hydrocarbon group having a degree of unsaturation of 0-2 and 12-24 carbon atoms, R2 is an

Description

Title of the Invention: Composition and use thereof Technical Field

[0001] The present invention relates to a composition comprising a diacyl glyceryl ether (DAGE) and use thereof. Background Art

[0002] Diacyl glyceryl ethers (DAGEs) are widely distributed in marine animal and fish fats and oils, and are particularly abundant in fats and oils derived from shark and squid livers. Several functions are known to DAGEs. For example, Patent document 1 describes an oral agent for prevention and treatment of skin damage caused by ultraviolet rays, containing a diacyl glyceryl ether as an active ingredient. Patent document 2 describes a composition for improving the quality of sleep, containing a fish egg lipid preparation and a diacyl glyceryl ether (DAGE).

[0003] For DAGEs, there have also been reported effects on circadian rhythms (Nonpatent document 1), and that intake of salmon roe oil containing DHA and EPA and deep-sea shark liver oil containing DAGE by healthy people who were temporarily dissatisfied with their sleep was confirmed to increase the ratio of deep sleep and REM sleep, and thereby improve the quality of sleep, further, alleviated anxiety, tension, and dejected mood, improved vigor and activity, and reduced daytime sleepiness in those with decreased vigor and activity (Non-patent documents 2 and 3). Furthermore, the effect of intake of foods containing deep-sea shark liver oil by Japanese healthy subjects on their activity has been reported (Non-patent document 4). Prior Art References Patent documents

[0004] Patent document 1: International Publication WO2004 / 000301 (Japanese Patent No. 4417249) Patent document 2: International Publication WO2022 / 210856 Non-patent documents

[0005] Non-patent document 1: Journal of Chronobiology, Vol. 28, No. 2, p216, 2022 Non-patent document 2: Nakagawa, Iku et al., Verification of the sleep quality improving effect and anti-stress effect of a processed food product containing salmon roe oil-derived DHA and EPA and deep-sea shark liver oil-derived DAGE in healthy adults who are dissatisfied with their sleep - A randomized, double-blind, placebo-controlled, parallel-group study, Jpn Pharmacol Ther (Yakuri to Chiryo), Vbl.49, No.6, p977-990, 2021 Non-patent document 3: Takada, Yuka et al., The frontline of the development of foods with functional claims for fatigue, stress and sleep - REM sleep and deep sleep increasing action of oil and fat processed foods containing DHA, EPA, and DAGE (diacyl glyceryl ether), Monthly Fine Chemicals, Vol. 52, No. 4, p40-48, 2023 Non-patent document 4: Yoshinaga, Ikuo, Influence of the intake of foods containing deep-sea shark liver oil on the activity of Japanese healthy subjects: A randomized, double-blind, placebo-controlled, parallel-group study, Jpn Pharmacol Ther (Yakuri to Chiryo), Vol.46, No.9, pl399-1425, 2021 Summary of the Invention Object to be achieved by the invention

[0006] It would be desirable to provide a highly effective novel composition comprising DAGE. It would also be desirable if novel applications of DAGE could be provided. Means for achieving the object

[0007] The present invention provides the following. [1] A composition comprising: A. a diacyl glyceryl ether (DAGE) represented by the formula I:

[0008] [Formula I] ac-o-Ri i R2-O-CH ] HaC-o-R8 Formula I

[0009] (in the formula I, R1 is an aliphatic hydrocarbon group having a degree of unsaturation of 0 to 2 and 12 to 24 carbon atoms, R2 is an acyl group having a degree of unsaturation of 0 to 6 and 12 to 24 carbon atoms, and R3 is an acyl group having a degree of unsaturation of 0 to 6 and 12 to 24 carbon atoms), and B. squalene, wherein A:B (mass ratio) is 0.9 to 9:1. [2] The composition according to 1, which is prepared so that 400 mg or more of DAGE is ingested per day. [3] A composition for use in a therapeutic method of improving at least one selected from negative mood state, anger, fatigue, inertia, confusion, and mental health in a subject, wherein the composition comprises a diacyl glyceryl ether (DAGE) represented by the formula I:

[0010] [Formula I] ac-o-Ri i R2-O-CH ] HaC-o-R8 Formula I

[0011] (in the formula I, R1 is an aliphatic hydrocarbon group having a degree of unsaturation of 0 to 2 and 12 to 24 carbon atoms, R2 is an acyl group having a degree of unsaturation of 0 to 6 and 12 to 24 carbon atoms, and R3 is an acyl group having a degree of unsaturation of 0 to 6 and 12 to 24 carbon atoms). [4] The composition for use of 3, wherein the composition is for relieving at least one selected from anger, fatigue, inertia, and confusion in a subject in a negative mood state to improve the mood state. [5] The composition for use of 3 or 4, wherein the composition further comprises squalene. [6] The composition according to 1 or 2, or the composition for use of any one of 3 to 5, wherein the composition further comprises one or more triglycerides. [7] The composition according to 1 or 2, or the composition for use of any one of 3 to 6, which is for ingestion by a subject who easily feels stress or is in a negative mood state on a daily basis. [8] The composition according to 1 or 2, or the composition for use of any one of 3 to 7, which is for ingestion of 400 mg or more of DAGE per day for 12 weeks or longer. [9] The composition according to 1 or 2, or the composition for use of any one of 3 to 7, which is for ingestion of 400 mg or more of DAGE per day for 12 weeks or longer continuously.

[10] Anon-therapeutic method of improving at least one selected from negative mood state, anger, fatigue, inertia, confusion, and mental health in a subject, wherein the method comprises administering a composition comprising a DAGE as defined in 3.

[11] The method of 10, wherein the method comprises one or more of the features recited in 4-9. Effects of the Invention

[0012] As one aspect of the present invention, a composition comprising a diacyl glyceryl ether and squalene is provided. As another aspect, there is provided a composition for improving at least one selected from total mood disturbance, angerhostility, fatigue-inertia, confusion-bewilderment, and mental health, which comprises a diacyl glyceryl ether (DAGE). Modes for Carrying out the Invention

[0013] For the present invention, a numerical range represented as x to y includes the values x and y at both ends. For the present invention, numerical values of ratios and proportions are based on mass, unless especially noted.

[0014] [Composition] The first aspect of the present invention is a composition comprising a diacyl glyceryl ether (DAGE) and squalene.

[0015] (Diacyl glyceryl ether) The composition of this aspect comprises a diacyl glyceryl ether (DAGE) represented by the formula I. DAGE is also referred to as alkyldiacylglycerol. DAGE has a structure that fatty acids are ester-bonded to two of the three hydroxyl groups of glycerol (glycerin) and a hydrocarbon is ether-bonded to the remaining one hydroxyl group, and represented by the following formula I.

[0016] [Formula I] ac-o-Ri i R2-O-CH ] HaC-o-R8 Formula I

[0017] In the formula I, R1 is an aliphatic hydrocarbon group having a degree of unsaturation of 0 to 2 and 12 to 24 carbon atoms, R2 is an acyl group having a degree of unsaturation of 0 to 6 and 12 to 24 carbon atoms, and R3 is an acyl group having a degree of unsaturation of 0 to 6 and 12 to 24 carbon atoms.

[0018] Example of the aliphatic hydrocarbon group represented by R1 include, for example, those of which carbon atom number and degree of unsaturation are 24 and 1, 22 and 1, 20 and 1,18 and 2, 18 and 1,18 and 0, 16 and 1, 16 and 0, 14 and 0, 12 and 0, and so forth. Those of which carbon number and degree of unsaturation ratio are 18 and 1,18 and 0, 16 and 1, or 16 and 0 are particularly preferred.

[0019] Examples of the acyl group represented by R2 include, for example, those of which carbon atom number and degree of unsaturation are 24 and 1 (derived from nervonic acid), 22 and 6 (derived from docosahexaenoic acid), 22 and 1 (derived from erucic acid), 20 and 5 (derived from eicosapentaenoic acid), 20 and 4 (derived from arachidonic acid), 20 and 1 (derived from gadoleic acid), 18 and 3 (derived from a-linolenic acid or y-linolenic acid), 18 and 2 (derived from linoleic acid), 18 and 1 (derived from oleic acid or octadecenoic acid), 18 and 0 (derived from stearic acid), 16 and 1 (derived from palmitoleic acid), 16 and 0 (derived from palmitic acid), 14 and 0 (derived from myristic acid), 12 and 0 (derived from lauric acid), and so forth.

[0020] Those of which carbon atom number and degree of unsaturation are 24 and 1 (derived from nervonic acid), 22 and 6 (derived from docosahexaenoic acid), 22 and 1 (derived from erucic acid), 20 and 5 (derived from eicosapentaenoic acid), 20 and 1 (derived from gadoleic acid), 18 and 1 (derived from oleic acid or octadecenoic acid), or 16 and 0 (derived from palmitic acid) are particularly preferred.

[0021] Examples of the acyl group represented by R3 include, for example, those of which carbon atom number and degree of unsaturation are 24 and 1 (derived from nervonic acid), 22 and 6 (derived from docosahexaenoic acid), 22 and 1 (derived from erucic acid), 20 and 5 (derived from eicosapentaenoic acid), 20 and 4 (derived from arachidonic acid), 20 and 1 (derived from gadoleic acid), 18 and 3 (derived from a-linolenic acid or y-linolenic acid), 18 and 2 (derived from linoleic acid), 18 and 1 (derived from oleic acid or octadecenoic acid), 18 and 0 (derived from stearic acid), 16 and 1 (derived from palmitoleic acid), 16 and 0 (derived from palmitic acid), 14 and 0 (derived from myristic acid), 12 and 0 (derived from lauric acid), and so forth.

[0022] Those of which carbon atom number and degree of unsaturation are 24 and 1 (derived from nervonic acid), 22 and 6 (derived from docosahexaenoic acid), 22 and 1 (derived from erucic acid), 20 and 5 (derived from eicosapentaenoic acid), 20 and 1 (derived from gadoleic acid), 18 and 1 (derived from oleic acid or octadecenoic acid), or 16 and 0 (derived from palmitic acid) are particularly preferred.

[0023] DAGEs are known to be abundantly comprised in liver oil of sharks, especially deep-sea sharks. In addition to sharks, they are also known to be comprised in squid (liver), some fish (muscle), cattle, human (milk), and silver chimaera (silver shark, Chimaera phantasma). The raw material of DAGE used in the composition of this aspect of the present invention is not particularly limited. For the composition of this aspect of the present invention, as DAGE, shark liver oil may be used, or concentrated, crude, or purified shark liver oil may be used. One produced by microorganisms or a synthetic product may also be used.

[0024] In one embodiment, DAGE may be one comprised in deep-sea shark liver oil. Deep-sea shark liver oil is extracted from the liver of sharks that can live in the deep sea, such as dwarf gulper shark (Centrophorus atromarginatus), roughskin dogfish (Centroscymnus owstonii), and birdbeak dogfish (Deania calceus). Liver oil can be extracted by conventionally known methods. For example, shark liver can be taken out, crushed into small pieces using a crusher, and left to stand for 1 to 4 days, the oil and solid contents can be separated, and the oil content can be extracted to obtain liver oil. The liver oil may be further purified by filtering to remove fine solids. Alternatively, commercially available deep-sea shark liver oil can also be used. Immune activating functions have been reported for alkylglycerols comprised in shark liver oil (Mar. Drugs 2010, 8, 2267-2300; doi:10.3390 / md8082267, and Lipids Health Dis. 2014 Nov 27:13:178. doi: 10.1186 / 1476-511X-13-178)

[0025] (Squalene) The composition of this aspect of the present invention comprises squalene. Squalene (IUPAC name: 2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene) is an oil substance belonging to triterpenes.

[0026] Squalene is abundantly comprised in shark liver oil and is also found in vegetable oils, such as olive oil, rice bran oil, wheat germ oil, sesame oil, and soybean oil. Algae producing squalene are also known (Japanese Patent Nos. 6265407 and 5942197).

[0027] The raw material of squalene used in this aspect of the present invention is not particularly limited. In one embodiment, the oils mentioned above may be used as squalene, or concentrated, crude or refined products of the oils mentioned above may be used. Antioxidation effects have been reported for squalene, and the inclusion of squalene in the composition is expected to support the action of D AGE at the time of ingestion and in the body.

[0028] (Mass ratio and content) The mass ratio of A (DAGE) and B (squalene) in the composition of this aspect of the present invention can be selected as appropriate. In one embodiment, A:B can be 0.1 to 10:1, e.g., 0.8 to 10:1 or 0.1 to 9:1, preferably 0.9 to 9:1, more preferably 1 to 8:1, more preferably higher than 1 to 7.5:1 (DAGE rich), further preferably 2 to 7:1. The A:B ratio in commercial functional foods comprising DAGE is analyzed to be 0.07 to 0.77:1.

[0029] The content of DAGE in the composition of this aspect of the present invention can be appropriately selected, and in one embodiment, it may be 38 g or more per 100 g of the composition (when the composition is comprised in capsules, or the like, per 100 g of mass including the mass of such coatings as capsules, the same shall apply hereinafter), preferably 39 g or more, more preferably 40 g or more, further preferably 41 g or more, or may be 42 g or more, 43 g or more, 44 g or more, or 45 g or more. The content of DAGE in the composition can be analyzed as total alkoxyglycerol content after saponifying DAGE. In one embodiment, the content of DAGE per 100 g of the composition can be 14 g or more, 15 g or more, 16 g or more, 17 g or more, or 18 g or more as the total alkoxyglycerol when saponified.

[0030] In another embodiment, the amount of DAGE in the composition can be 1 to 50 g, preferably 16 to 40 g, more preferably 18 to 35 g, further preferably 20 to 30 g, per 100 g of the composition.

[0031] The content of squalene in the composition of this aspect of the present invention can be selected as appropriate, and in one embodiment, it can be 1 to 50 g, preferably 2 to 40 g, more preferably 3 to 30 g, further preferably 5 to 20 g, per 100 g of the composition.

[0032] (Others) In one embodiment, the composition may comprise an ingredient other than DAGE and squalene. Examples of such an ingredient include triglycerides. Triglyceride is a compound consisting of three molecules of fatty acid ester-bonded to one molecule of glycerol, and may be also called triacylglycerol or triacyl glyceride. In one embodiment, when DAGE in the composition is analyzed by saponification, alkoxyglycerols (e.g., chimyl alcohol, batyl alcohol, and selachyl alcohol) are found.

[0033] If the composition comprises one or more triglycerides, the amount thereof can be appropriately selected, and in one embodiment, it may be 1 to 50 g, preferably 2 to 40 g, more preferably 3 to 30 g, further preferably from 5 to 30 g, per 100 g of the composition.

[0034] In one embodiment, the total amount of alkoxyglycerols in the composition when saponified is 14.5 to 50 g, preferably 14 to 40 g, more preferably 15 to 35 g, further preferably 16 to 30 g, per 100 g of the composition. More precisely, content of chimyl alcohol is 2 to 18 g, preferably 2.2 to 16 g, more preferably 2.6 to 13 g, further preferably 3 to 10 g, per 100 g of the composition. Content of batyl alcohol is 1.5 to 18 g, preferably 1.7 to 16 g, more preferably 1.9 to 13 g, further preferably 2.1 to 10 g, per 100 g of the composition. Content of selachyl alcohol is 6.4 to 18 g, preferably 6.7 to 16 g, more preferably 7 to 13 g, further preferably 7.3 to 10 g, per 100 g of the composition.

[0035] The composition of this aspect of the present invention can be used for the use of the second aspect of the present invention explained below.

[0036] [Use] The second aspect of the present invention is a composition for improving at least one selected from total mood disturbance, anger-hostility, fatigue-inertia, confusion-bewilderment, and mental health, which comprises DAGE as an active ingredient.

[0037] (Active ingredient) The active ingredient used in this aspect of the present invention is DAGE. To DAGE, all the above explanations of DAGE for the first aspect can be applied as they are. A combination of DAGE and squalene may be used as the active ingredient. To squalene, all the above explanations of squalene for the first aspect can be applied as they are.

[0038] (Action and function) The composition of this aspect of the present invention can be used to improve at least one selected from negative mood state, anger, fatigue, inertia, confusion, and mental health. Anger referred to here can also be expressed as irritation (state of anger), hostility or antagonism, fatigue or inertia as feeling of fatigue or feeling of inertia (state of being exhausted), and confusion as a state of bewilderment or embarrassment (state of being unable to think clearly), etc. The negative mood state comprehensively represents state of depressed mood or mental disorder state such as anger, hostility, fatigue, inertia, confusion, bewilderment, tension, anxiety (uncomfortable state of mind), depression (state of dejected mood), dejection (state of depressed mind), etc.

[0039] The negative mood state, anger, fatigue (inertia), and confusion can be evaluated as Total Mood Disturbance (TMD), Anger-Hostility (AH), Fatigue-Inertia (FI), and Confusion-Bewilderment (CB), respectively, together with Tension-Anxiety (TA), Depression-Dejection (DD), Vigor-Activity (VA), and Friendliness (F) according to "POMS (registered trademark) 2 Japanese Edition (Kaneko Shobo, Ltd.)" and "POMS 2 Japanese Manual (Kaneko Shobo, Ltd.)". According to POMS 2, mood state over a given time frame is evaluated with seven scales: anger-hostility, fatigueinertia, confusion-bewilderment, tension-anxiety, depression-dejection, vigor-activity, and friendliness, as well as the "TMD score", which comprehensively represents negative mood state.

[0040] Specifically, a user can answer the prescribed 6 to 12 questions for each item on a 5-point scale from 0 to 4, and evaluation for each item can be performed by using the sum of the scores for each item as the prime score, and the TMD score can be calculated by subtracting the prime score for VA from the sum of the prime scores for AH, CB, DD, F, and TA to evaluate the TMD. AH (anger-hostility) indicates anger and antipathy toward others, intense anger, as well as inner annoyance and desire to be mean to others. CB (confusion-bewilderment) indicates bewilderment and low cognitive efficiency, or state of confusion and inability to think clearly DD (depression-dejection) indicates state of loss of self-confidence, such as feeling oneself worthless, hopeless, or guilty FI (fatigue-inertia) indicates state of feeling fatigue and decreased awareness and activity. TA (tension-anxiety) indicates increased tension and anxiety, characterized by heightened nervousness and restlessness. VA (vigor-activity) indicates high energy, dynamism, and activity. F (friendliness) indicates a positive mood state, and interpersonal influences.

[0041] Mental health (MH) can be assessed with the MOS Short-Form 36-Item Health. Survey (SF36v2) Japanese version (distributor Qualitest Co., Ltd.).

[0042] Specifically, mental health can be assessed by answering questions about how a user consider his or her own health on a 5-point scale from 1 to 5 using a prescribed questionnaire. The questions relate to health status; current health status compared with one year ago; difficulties in daily activities; physical or psychological problems in doing work or usual activities (e.g., housework) in the past one month; whether socializing with peers was disturbed by physical or psychological reasons; physical pain, whether this disturbed usual work, and so forth (J Clin Epidemiol Vol. 51, No. 11, pp. 1037-1044, 1998; J Clin Epidemiol Vol. 51, No. 11, pp. 1045-1053, 1998).

[0043] In one embodiment, the composition is suitable for use to relieve at least one selected from anger, fatigue, inertia, and confusion in a subject in a negative mood state, i.e., at least one selected from anger-hostility, fatigue-inertia, and confusionbewilderment in a subject in a depressed mood state, to improve the mood state. The negative mood state or depressed mood state may be temporary or persistent, but when the composition is used as a food, it is preferably temporary in view of the function of such a food.

[0044] In one embodiment, the composition can also be used to improve at least two or more selected from total mood disturbance, anger-hostility, fatigue-inertia, confusionbewilderment, and mental health. In a preferred embodiment, the composition can be used to improve three or more or four or more selected from them. In a particularly preferred embodiment, the composition can be used to improve all of them.

[0045] A subject in a depressed mood state is a subject whose TMD is higher than 50, more preferably 52 or higher, further preferably 55 or higher, as assessed by POMS2, for example.

[0046] (Subject) The composition of this aspect of the present invention is suitable for ingestion by a subject for whom it is desirable or necessary to improve at least one selected from negative mood state, anger, fatigue, inertia, confusion, and mental health, that is, a subject who has been rated low for at least one of negative mood state, anger, fatigue, inertia, confusion, and mental health.

[0047] In one embodiment, the composition is suitable for ingestion by a subject who easily feels fatigue and stress on a daily basis. In another embodiment, the composition is suitable for ingestion by a subject who has shown a relatively high TMD score in POMS2.

[0048] The subject may be a human or a non-human animal. In one embodiment, the subject is preferably a healthy subject (e.g., not diagnosed as sick by a physician). Such a healthy subject may be, for example, a person who does not correspond to: - a person who is under a treatment for or has a history of malignant tumor, heart failure, or myocardial infarction, - a person with an implanted pacemaker or implantable cardioverter defibrillator, - a person under a treatment for any of the following chronic diseases: arrhythmia, hepatic disorder, renal disorder, cerebrovascular disorder, rheumatism, diabetes, dyslipidemia, hypertension, and other chronic diseases, - a person with a history of a psychiatric disorder such as depression and attention-deficit / hyperactivity disorder (ADHD), and - a person under a treatment for chronic fatigue syndrome or menopausal disorders.

[0049] In addition to the above, a healthy subject may also be defined as a person who does not correspond to: - a person who regularly use a pharmaceutical (including herbal medicine) or dietary supplement, - a person who regularly consume a food for specified health use or food with functional claims, - a person with an allergy (to drug or related food) - a pregnant or lactating woman, - a person who has irregular sleeping hours or sleeping habits due to night shifts, etc., - a person with irregular lifestyle habits (eating habits, exercise habits, sleeping habits, etc.), and - a person engaged in physical labor such as carrying heavy objects.

[0050] In one embodiment, the compositions can be used for a non-therapeutic purpose. To use for a non-therapeutic purpose means to use the composition for a purpose other than treating a disease.

[0051] Such a subject as described above may be an adult (15 years of age or older for humans), middle-aged person, or elderly person (65 years of age or older for humans). In a preferred embodiment, the composition is suitable for ingestion by a subject who is 30 years of age or older and younger than 70 years of age.

[0052] (Others) The composition of this aspect of the present invention may comprise an ingredient other than DAGE as the active ingredient. Examples of the ingredient other than DAGE include squalene and triglycerides. To squalene and triglycerides, all the explanations for them made for the first aspect of the present invention are applied as they are.

[0053] [Form of composition, etc.] (Food composition and pharmaceutical composition) The compositions of the first aspect and the second aspect of the present invention (hereinafter both may be collectively referred to as the composition of the present invention) can be a food composition or pharmaceutical composition. Foods include general foods, functional foods, and nutritional compositions, unless especially noted. The foods also include not only solids, but also liquids, e.g., beverages, drinkable preparations, and soups, unless especially noted. The functional foods are foods that utilize the functions of ingredients, and include all types of health foods, such as foods for specified health uses, foods with functional claims, foods with health claims including foods with nutrient function claims, foods for special dietary uses, nutritional supplement foods, health supplement foods, supplements, and cosmetic foods.

[0054] (Administration route) The composition of the present invention can be orally ingested. For the present invention, ingestion is used in the sense of giving a food composition to a subject as well as administering a pharmaceutical composition to a subject. Ingestion (to make someone ingest) can be read as administration (to administer).

[0055] (Content and dose of active ingredient) The content of the active ingredient in the composition of the present invention may be such an amount that the desired effect is exhibited. The content of the active ingredient in the composition of the present invention can be appropriately set in consideration of various factors such as the age, weight, and symptoms of the subject of the ingestion or administration.

[0056] In one embodiment, the composition of the present invention can be prepared so that the daily ingestion amount of the active ingredient is, for example, 50 mg or more, or may be prepared so that the daily ingestion amount of the active ingredient is 100 mg or more, 150 mg or more, 200 mg or more, 250 mg or more, 300 mg or more, or 350 mg or more. In a preferred embodiment, the composition can be prepared so that the daily ingestion amount of the active ingredient is 360 mg or more, further preferably 400 mg or more. The maximum amount can be defined as appropriate, and the amount may be, for example, 1800 mg or less, 1000 mg or less, or 500 mg or less, no matter how the minimum amount is defined. One unit of the composition may be ingested by a subject once a day, or may be ingested by a subject multiple times a day, e.g., three times a day. In one embodiment, if the composition of the present invention comprises squalene in addition to the active ingredient, it can be prepared so that the daily ingestion amount of squalene is, for example, 5 mg or more, or may be prepared so that the daily ingestion amount of squalene is 10 mg or more, 20 mg or more, 50 mg or more, 100 mg or more, or 150 mg or more. The maximum amount can be appropriately defined, and the amount may be, for example, 1000 mg or less, or 500 mg or less, no matter how the minimum amount is defined.

[0057] The content of the active ingredient per unit (e.g., one capsule) of the composition may be, for example, 10 mg or more, or may be 20 mg or more, 30 mg or more, 40 mg or more, or 50 mg or more, and is preferably 60 mg or more, more preferably 80 mg or more, further preferably 100 mg or more. The maximum content can be appropriately defined, and the content may be, for example, 500 mg or less, 400 mg or less, or 300 mg or less, no matter how the minimum content is defined. Several, for example, 1 to 9, 2 to 8, 3 to 7, or 4 to 6 single units of the composition may be ingested by a subject as a daily amount or amount for single time of ingestion, and the composition of such an amount may be ingested by a subject by ingestion of multiple times a day, e.g., 3 times a day, as divided portions.

[0058] Since the composition of the present invention consists of ingredients derived from natural products that have been eaten from old days, it may be ingested repeatedly or over a long period of time, e.g., 3 days or longer, 1 week or longer, or 2 weeks or longer, preferably 4 weeks or longer, more preferably 8 weeks or longer, e.g., 12 weeks. Such repeated or prolonged ingestion is preferably continuously performed everyday without any intervals.

[0059] (Other ingredients and additives) The composition of the present invention may comprise an ingredient acceptable for use in foods or pharmaceuticals. Examples of such an ingredient acceptable for use in foods or pharmaceuticals include lipids, proteins, amino acids, vitamins (e.g., vitamin A, vitamin Bl, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, biotin, folic acid, pantothenic acid and nicotinic acids), minerals (e.g., copper, zinc, iron, cobalt, and manganese), astaxanthin, and so forth. Astaxanthin has an action of protecting DAGE and squalene from peroxidation and stabilizing them.

[0060] The composition of the present invention may further comprise an additive acceptable for use in foods or pharmaceuticals. Examples of such an additive include excipients, surfactants, binders, disintegrants, lubricants, dissolution aids, suspending agents, coating agents, colorants, preservatives, buffers, pH adjusters, emulsifiers, stabilizers, sweeteners, antioxidants, flavorings, and acidifiers.

[0061] (Dosage form or shape) According to one embodiment, the composition of the present invention may be in any form, for example, in the form of soft capsule, hard capsule, tablet, coated tablet, sugar-coated tablet, and drinkable preparation.

[0062] (Others) In the manufacture of the composition of the present invention, the time of blending of the active ingredient may be appropriately selected, and is not particularly limited as long as the properties of the active ingredient are not significantly impaired.

[0063] According to one embodiment, the composition of the present invention can be labeled with an indication of purpose of use (intended use), functions of the active ingredient, and method of use based on the functions.

[0064] Specific examples of the indication include indications that the composition helps maintenance of mental health by relieving temporary anxiety, tension, fear, anger etc. at work or home; the composition supports maintenance of mental health by relieving temporary depressed mood state caused by anger, hostility, confusion, bewilderment, or inertia; the composition helps maintenance of mental health by improving temporary anxiety, feeling of fatigue, or feeling of inertia; the composition supports relief of temporary self-perceivable state of confusion (inability to think clearly), feeling of inertia (state of being exhausted), dejected state (state of depressed mind), irritation (state of anger), and anxiety (uncomfortable state of mind) in people who easily feel fatigue (and stress) on a daily basis, and so forth.

[0065] In one embodiment, the composition has an indication that administration of the composition is recommended to specific subjects. Examples of such subjects are as described above.

[0066] The indication may be direct or indirect indication. Examples of the direct indication include descriptions on tangible articles such as a product itself, package, container, label, and tag, and examples of the indirect (implied) indication includes advertising and campaign activities using such places or means as web site, shop, pamphlet, exhibition, seminar, book, newspaper, magazine, television, radio, postal matter, E-mail, and sound.

[0067] Hereafter, the present invention will be more specifically explained with reference to examples. Examples

[0068] [Example 1: Evaluation of functionality of DAGE] I. Methods 1. Purpose To examine anti-psychotic stress effects observed in healthy subjects who ingested a test food for 12 weeks.

[0069] 2. Design of study Objective of study: Efficacy and safety Type of study: Intervention Basic design: Parallel group comparison Randomization: Randomized Blinding: Double blind Control: Placebo control Number of study groups: 2 groups Number of studies: 3 times (before screening and enrollment, 6 weeks after enrollment, and 12 weeks after enrollment) Number of cases screened (*): 88 persons Target number of subjects: 44 persons Number of enrolled subjects: 48 persons *Number of cases screened: Minimum number of cases to participate in screening test for selection of subjects of the target number of subjects and the number of subjects to be enrolled.

[0070] 3. Eligibility criteria (1) Inclusion criteria The subjects for the study were selected from those who met the following criteria i to v, using the criteria of vi. i. Japanese ii. Man or woman iii. Thirty to and seventy years old iv. Healthy person v. Person who easily feels fatigue and stress on a daily basis. vi. Person who showed a relatively high TMD score in POMS2 among those who agreed to participate in the study

[0071] Subjects who did not meet the following exclusion criteria were considered healthy subjects. i. Person who is under a treatment for or has a history of malignant tumor, heart failure, or myocardial infarction ii. Person with an implanted pacemaker or implantable cardioverter defibrillator iii. Person under a treatment for any of the following chronic diseases: arrhythmia, hepatic disorder, renal disorder, cerebrovascular disorder, rheumatism, diabetes, dyslipidemia, hypertension, and other chronic diseases iv. Person who consumes a food for specified health use or food with functional claims on a daily basis. v. Person who regularly uses a pharmaceutical (including herbal medicine) or dietary supplement vi. Person with an allergy (to pharmaceutical or food relating to test food) vii. Woman who is pregnant, lactating, or intending to become pregnant during the study period viii. Person who have participated in other clinical trials during the 28 days prior to the date of obtaining consent, or who are scheduled to participate in other clinical trials during the study period ix. Person with a history of a psychiatric disorder such as depression and attention-deficit / hyperactivity disorder (ADHD) x. Person with irregular sleeping hours and sleeping habits due to night shifts, etc. xi. Person with irregular lifestyle habits (eating, exercise, sleeping habits, etc. xii. Person under a treatment for chronic fatigue syndrome or menopausal disorders xiii. Person engaged in physical labor such as carrying heavy objects xiv. Person judged by the investigator to be inappropriate for this study for other reasons.

[0072] 4. Intervention (1) Test food Test food group: Food containing shark liver oil Placebo group: Placebo (2) Involved ingredients Squalene and DAGE (diacyl glyceryl ether) (3) Test period Duration of intervention: 12 weeks (4) Administration and dose Test food group: Subjects consume 4 capsules per day with water or lukewarm water without chewing. Placebo group: Subjects consume 4 capsules per day with water or lukewarm water without chewing.

[0073] [Table 1] ( il’Olip I’eSl 1()()(1 1 )osage Ibrni ( Via! in capsule Material Rcrincd deep-sea shark liver oil 1 3 (.Manilla Nichiro Corporation) Squalene*3 (Maruha Nichiro Corporation) ( ollli'lll 2.-X) nig 50 mg Placebo Gelatin capsule Soybean oil*4 300 mg *1: 1000 mg (450 mg DAGE) per daily ingestion amount (4 capsules) *2: DAGE 112.5 mg, triglycerides 137.5 mg *3: 200 mg per daily ingestion amount (4 capsules) *4: 1200 mg per daily ingestion amount (4 capsules)

[0074] 5. Evaluated items (1) Profile of Mood States 2nd Edition (POMS2) Japanese Version i. Description: To assess current mood state of study participants by using the POMS2 Japanese version. ii. Survey items: Questionnaire items for each of total mood disturbance (TMD), tension-anxiety (TA), depression-dejection (DD), anger-hostility (AH), vigoractivity (VA), fatigue-inertia (FI), confusion-bewilderment (CB), and friendliness (F). iii. Evaluation method: Evaluation is performed by the test before screening and enrollment, test at 6 weeks after enrollment, and test at 12 weeks after enrollment. iv. POMS2 is "a test that allows quick assessment of relatively enduring affect states, as well as transient, fluctuating feelings", and the evaluation items thereof include seven factors: AH, CB, DD, FI, TA, VA, and F, and TMD (= AH + CB + DD + FI + TA - VA) as an indicator of total mood disturbance. The transformed (T) score used in the evaluation with POMS2 is a value calculated with normalized evaluation criteria, of which average is set to be 50 and standard deviation as 10. Lower T scores for TMD, AH, CB, DD, FI, and TA, which evaluate negative mood states, and higher T scores for VA and F, which evaluate positive mood states, indicate a better state (Heuchert JP, McNair DM. POMS2 Japanese version for adults - All items version. Tokyo: Kaneko Shobo; 2015).

[0075] (2) MOS Short-Form 36-Item Health Survey (SF36v2) i. Description: To assess health-related quality of life using the SF-36v2. ii. Survey item: Mental health (MH) iii. Evaluation method: Summary scores and subscale scores are calculated by scoring based on national standards. They are measured by the test before screening and enrollment and test at 6 weeks after enrollment.

[0076] 6. Outcome Efficacy evaluation items i. Primary outcome TMD, TA, DD, AH, VA, FI, CB, and F (POMS2) determined in the test at 6 weeks and tests at 12 weeks after enrollment ii. Secondary outcome MH determined in the test at 6 weeks after enrollment

[0077] II. Results Outline of the results (results for MH in POMS2 and SF36v2) is shown in the following tables.

[0078] [Table 2] Effective Primary outcome TMD Tension-anxiety (TA) POMS2 . . . Fatigue-inertia (FI) Confusion-bewilderment (CB) Secondary outcome SF36v2 Mental health (MH)

[0079] Placebogroup Item Unit Type Time point Year. 8S^®SB 1101 lOiii Sr —> Srrrg ’.l. / ^jrcdwsir; ......"s?T" ’"IsT....... VISIT2 Oi 24 55.2 8.2 55.0 40.0 69.0 55.0 1.1 $2.7 VISITS ¢. t 24 53.7 6.3 54.0 40.0 65.0 $3.6 1.2 51.2 .AH Score wasuredvisnr ■ 7 / 54.1 V: 2 7 77.8 7 / 7 :263.5- / 7741.077 7769.0 77 - 2777 - 7 7 - visit: = * 24 '3: / ;:S.5V -■ •s 0 V? 53' : / :21577 $0.6 visits - • 52 2 6.4 :7 741,0 2 2 66-0 7:752.722 : 3 7 / 250.177:2: 7CB Score Measuredvism • 1 24 59.6 8.5 59.0 41.0 73.0 VISIT’ Ml 24 57.3 9.1 ss.o 42.0 74.0 56.2 1.0 54.2 VISITS Oi 24 55.6 6.4 56.0 440 65.0 55.0 1.2 52.7 DD Score »asured visit) ■ - 7 55.9 ? <' OV 2 / :54.57::: 7745,077 - 7 / 7 / :- / / / / 7 “ 13 visit: . = -24 545 91 7 253.0 V 41.0 7277.071 '49 M 51.6 ■ VISIT? = » 24 2 / 52.31;? ¢6 2:53.077 2742.022 \ v / 7252.2772 1.1 ?■? V FI Score Measuredvism * i 24 5S.5 7.S 57.0 45.0 73.0 vism *7* $5.2 8.6 56.5 43.0 66,0 $5.0 1.3 $2.3 VISITS 4, t 7 24 53.4 7.7 54.0 41.0 71.0 53.2 1,3 50.5 TA Score MeasuredvBiTi * 24 2759.5227 22 8.477 22241-0 7 / : / 76.0 7; • 777 7- / / 777 visit: - * 24 T55.4V SA 2753.077: 7744.O777 27771,0 72 7755.0712 1.2 52 5 visits = = 24 ; : 53.52: 64 72 55-0 2 77740.077? 2 / 63-0 77 2253.3 2 2 1.3 27 / 50.87 2: 7 VA Score Measuredvism • : 24 46.6 6.7 45.0 38.0 62.0 \ ism = * - '2 ,0 $ -Ce O' / .3 ' 0 n6 6 VISITS * H 24 '6 37.0 6' ’ -3 2 1 4 45.1 F Score MeasuredvMTi • 24 69 -: 2 27 7-7 / 7: - 777 77- 7 / 7 77 W U° VISIT? = - 2» 510 5 tv 3'0 03 2 '0 ? 1 3 VISITS = ' 24 40? '6 -.3 3 69 C AS ’ 6 -4 s MH Score Measuredvieir: IlSAlaAT a? 4 '8 4: «5 3-3 5' 1 - 777777 / -17 / 771 7 / / / / VTS1I2 = 24 48 4 49 > 36 8 62 : «3 1 3 40 4 ““7 Group comparison isoww ?T 2-s $7.6 6.2 Lv -4.0 70 0 -0.6 19 -4 4 3 2 0.742 U) $7,2 24 53.2 6.5 52.5 42..0 63.0 $3.4 LI 51.2 55.? -1.5 1.6 -4.7 1.6 0.340 55.9 23 6.4 48.0 38.0 6L.0 49.6 1.2 <20 4$ 17' -0.6 7 / / -777 :7-724 7 ^56.7^ 9.5 / / / : / 55.5 / / / / / / 39.0 / / / / / / 71.0 / / / - - - : 7 / 7- / / 7 / :: 7 7 / / :2.6 / / / : : 7 / 2. 5 7 / / / / / -2.5 / / / / / : / : / / 7,7 / 7 / : / / 0.303 7 >6' ' / / / 24 / : 7 / 50.97 7 '0 33. / / / 71.0 / / : / / / 50.4 / / L5 4' * :3 4 3 3 2 1 -"6 1.0 ■ 131 55 2 23 749.0 / / / / / 749.0 / / / / : / 38.0 / / / / / / 64.0 / : / 43.8 77 1 5 7 / / 46.2 / / / :7 / 51.4 / 7: / -3 9 • S 5 / : / -0.2 / / 0.0« 24 57.1 6.8 57.5 42.0 68.0 -2.5 2.2 ■7.0 2.0 0.266 58.2 24 $3.3 6.4 53.0 42.0 68.0 54.3 1.0 $2.2 56.3 -L9 1.4 -4.8 1.0 0.IS6 57.4 23 49.7 6.S 49,0 36.0 64.0 50.4 1.2 48.0 52.8 -4.6 1.7 -8.0 -L2 24 : / / 54.3 / / / 7 / / 5.7 / / / / / / 754.5 / / / / / / / 42,0 / / / 69.0 ■■■ - 2.0 : / : / -5.7 / / / : / / 2.4 / / :: / / 0.411 / 7 56.1 2- '' / / / / 40.0 / / / '2 ’ l.I / / / / / 50.5 / / / ^5 0 1.6 / 7 / :-4.4 7 7 2 ; 54: 23 / / 49.2 / 7 / 69 46 0 40.0 7: / 62..0 / / / 7 / 50.0 / 7 / / / 47.7 / / : 7 / :52.37 / 7 -2.2 1 6 -54 : / / / 1.0 / 77 / 0.169 / / : 24 57.6 95 56.0 39.0 77.0 -6:.9 2.5 -6.0 4.1 0.717 57.6 24 52.2 7.6 53.0 39.0 66..0 5? 5 1 3 49.8 55 2 -26 1.9 -6.3 1.3 0.194 55.9 23 46.7 7.6 45.0 36.0 59.0 47.1 1.3 44.4 49.8 -6.1 L9 -9.9 -.: 5 ■• : / 7: 24 7 / / 58.0 / / 92 7 / 7 / 57.0 / 7 / / / / / 44.0 / / / / / 80.0 / / / 77 / -7 / : / • 7 / / -: 7 7 / / : I > 25 6 • : / 7 / 3.6 / / / : / 0.548 / / / / 57.5:7: 24 53" 7 / :6.8 / 7 7 / 7 54,0 7 / 41 0 / / / 67.0 54.1 1 2 : / : / 51.6 / / :756.67 / / / : :7 / / 7-0.9 / / / 1.8 / : / / -4.4 / / / 2.7 / / / / 0.619 / / : :55.9 / / / 23 / / / 49.4 / / / / : / / / 6.4 / / / 50 / / 310 / 1 / / / / 59.0 / / / 7 49.8 7 / 1.3 4'3 / ::52.4 / / / / : -3.5 : / / / 1.8 / / •' 1 / / / a i 7 / / / / 70.055 / / - 24 45.0 7.0 44.5 33.0 58.0 -1.7 2.0 •5.6 2.3 0.403 so.s 24 46.0 8.3 44.0 32.0 67.0 46.7 1.0 44.7 48.8 -1.9 1.5 -4.9 1.0 0.196 50.9 23 48.1 9.9 50.0 33.0 76.0 48.7 1.4 45.8 $1.6 0.7 2.0 -3.4 4.8 0.724 7 / :-7 / L 2- 8b x' 2 N< - - 7: / / 7-: / / / 7 / / ::- / 7 / / / / •3.4 2.3 3> "2 24 / / / 50.0 / / / / 1-. 2 4$, 7 / 37,0 / / / / / / 71.0 / / / 1 3 *8 5 . 9 I 9 'V .: 7 :51,3 / / 25 7 / 51.1 / / / : 'C^ / / 30.0 / / : / / / / 74.0 / / / / / / 52.1 / / / / 4 4 --. 4.1 2.3 -0.6 8 ' 18' • 2i '6 2 / / / / 57.:1 / / / - / 7: / / -7 : 7 7 I i / / / 2.0 / / / -.2.9 / / : / 5.0 / 7 0.595 : / : 24 ' :7 / / 7.0 / / 7 / 7 / / 47.0 / / / 2^ - / / 757,1 7 / 745.3 7 7- 7 / / / 1.3 / / / 7 742.77 / / ::747.9 / / : / •37'' ’ 8 : / :-7.477 / 770.0 7

[0080] n, Number of subjects; Mean, mean; SD, standard deviation; Med, median; Min, minimum value; Max, maximum value; EMM, estimated marginal mean; A, difference between groups (food group - placebo group); SE, standard error; 95%CI-, 95% confidence interval lower limit; 95%CI+: 95% confidence interval upper limit; P, significance probability; f, primary outcome; VISIT1, at test before screening and enrollment (baseline); VISIT2, at test 6 weeks after enrollment; VISIT3: at test 12 weeks after enrollment; *, Welch's t-test; #, linear mixed model using baseline value as covariate, and time point, group, interaction between time point and group, interaction between baseline value and time point, and study participants as factors (A is difference of estimated marginal means between groups); and orange box, significant difference (P <0.05)

[0081] On the basis of the above results, it is suggested with clinical significance that consumption of the test food for 12 weeks contributes to reduction of mental stress, in particular, improves TMD (general indicator of mood disturbance, emotional or psychological distress, and subjective well-being), AH (state of anger and antipathy toward others), FI (fatigue, inertia, and decreased activity), and CB, indicating negative mood states. It is also suggested that consumption of the test food for 6 weeks improves mental health.

[0082] [Example 2: Analysis of DAGE] (1) Equipments and apparatuses Rotary evaporator Normal-phase solid-phase extraction column High performance liquid chromatography complete set Charged particle detector

[0083] (2) Reagents 2,2,4-Trimethylpentane (isooctane, for high performance liquid chromatography) tert-Butyl methyl ether (for high performance liquid chromatography) Formic acid (for LC / MS) Chloroform (special grade) n-Hexane (for high performance liquid chromatography)

[0084] (3) Operation - Standard sample l,2-Dipalmitoyl-3-O-hexadecyl-rac-glycerol was used as the standard sample. This substance was synthesized by the following method. Under a nitrogen atmosphere, to 3-(hexadecyloxy)-1,2-propanediol (89.5 mg, 0.28 mmol) dissolved in dehydrated dichloromethane (6 mL), palmitic acid (201.5 mg, 0.79 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide (124.3 mg, 0.80 mmol), and 4-dimethylaminopyridine (8.0 mg, 0.066 mmol) were added, and the mixture was stirred at room temperature for 1.5 hours. The reaction mixture was concentrated under reduced pressure, applied to a normal-phase silica gel column (5 g), and purified with hexane-ethyl acetate (50:1), and the solvent was removed to obtain a white solid (150.1 mg, 0.19 mmol, yield 67%, purity 97.63% (purity was determined by quantitative NMR method)). The obtained 1,2-dipalmitoyl-3-O-hexadecyl-rac-glycerol was precisely weighed, dissolved in isooctane, and used as a standard for DAGE quantification. From the peak area values detected with the HPLC-charged particle detector and the concentrations, a calibration curve (regression equation represented by a quadratic equation: y = ax2 + bx + c) was created.

[0085] - Method for preparing sample solution (test food used in Example 1) The capsule was cut open, approximately 10 mg of the content was weighed, hexane was added, and the volume was fixed in a 10-mL volumetric flask. The solution was further diluted 50-fold with hexane, and subjected to HPLC analysis, and the concentration of DAGE was calculated from the calibration curve of the standard.

[0086] - HPLC conditions HPLC system: Nexera X2 (Shimadzu) Detector: Charged particle detector (Thermo Fisher SCIENTIFIC) Column: SunShell HILIC-S, 2.6 pm, 2.1 x 150 mm (ChromanNik Technologies Inc.) Mobile phase: A, 2,2,4-Trimethylpentane; B, tert-butyl methyl ether containing 0.01% (v / v) formic acid Flow rate: 0.6 mL / minute Gradient: B cone. 0.5% (0 to 1.5 minutes), 0.5 to 1.5% (1.5 to 1.51 minutes), 1.5 to 11% (1.51 to 7.5 minutes), 25% (7.5 to 10 minutes), 25 to 0.5% (10 to 10.5 minutes), and 0.5% (10.5 to 20 minutes) Column oven: 35°C Injection volume: 1 pL

[0087] As a result, the DAGE standard showed a peak around a retention time of 4.7 minutes. The test food used in Example 1 also showed a peak around a retention time of 4.7 minutes.

[0088] [Example 3: Analysis of DAGE content of commercial products] The DAGE contents of commercial products were determined by using the method of Example 2. The results are shown in the following table.

[0089] [Table 4] Squalene DAGE content Total alkoxyglycerols Chimyl alcohol Batyl alcohol Selachyl alcohol Test Food used in 18.7 45.6 18.1 3.4 2.3 7.8 Example 1 Commercial product A Commercial product B 39.0 22.4 13.4 1.9 1.4 6.3 48.8 37.6 12.2 1.8 1.0 5.1 Commercial product C 85.0 6.3 Unanalyzable 0.6 0.4 1.3 g / 100 g g / 100 g g / 100 g g / 100 g g / 100 g g / 100 g