Composition for improving cognitive function

Abstract

The present disclosure provides a composition for improving cognitive function that comprises a fish roe lipid preparation. In one embodiment, provided is a composition for improving cognitive function that comprises a fish roe lipid preparation prepared so as to provide about 100 mg or more and less than 300 mg per day of DHA, wherein the cognitive function is the linguistic ability of a subject who takes regular exercise.

Description

Composition for improving cognitive function

[0001] This patent application claims priority to Japanese Patent Application No. 2021-082910, the entire contents of which are incorporated herein by reference. The present disclosure relates to compositions for improving cognitive function.

[0002] The proportion of Japan's population aged 65 and over has been rising since 1950, reaching 28.4% in 2019. This proportion is expected to continue rising, reaching 35.3% in 2040, when the generation born during the second baby boom (1971-1974) will be 65 or older. With the rapid aging of society in recent years, the discrepancy between average life expectancy and healthy life expectancy is expected to widen, leading to a decline in quality of life and a longer period of need for care. Dementia, a particular concern among the elderly, is a serious social issue. As of 2012, the number of people with mild cognitive impairment (MCI), which is considered a precursor to dementia, was estimated to be approximately 4 million nationwide, meaning that approximately one in four elderly people has dementia or MCI. Mental health is also important for maintaining the physical and mental health of the elderly and preventing the need for care. It has been shown that when depression becomes severe and people become passive about managing their health and daily life, the condition and progression of various physical illnesses, such as ischemic heart disease such as myocardial infarction, cerebrovascular disorders such as stroke, diabetes, and cancer, worsens.

[0003] Currently, anti-dementia drugs on the market do not provide a fundamental cure, and new drug development is still ongoing around the world. There is also growing interest in preventing dementia, and it is considered important to take measures from the stage of age-related forgetfulness.

[0004] n-3 polyunsaturated fatty acids, such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), have attracted attention in recent years. Research has suggested that their intake may improve cognitive function (Non-Patent Document 1), and research is also underway to reduce the risk of developing Alzheimer's disease (Non-Patent Document 2). Traditionally, DHA and EPA derived from fish oil have been widely used. Meanwhile, salmon roe oil, rich in DHA and EPA, has been reported to increase REM sleep and improve sleep quality through oral ingestion (Non-Patent Document 3). Furthermore, lecithin is one of the raw materials for the neurotransmitter acetylcholine, and its ingestion is expected to improve cognitive function (Non-Patent Document 4). Furthermore, it has been suggested that DHA and EPA may be beneficial for improving mental health in elderly people with MCI (Non-Patent Document 5).

[0005] H Tokuda, et al., Low Doses of Long-Chain Polyunsaturated Fatty Acids Affect Cognitive Function in Elderly Japanese Men: A Randomized Controlled Trial. Journal of Oleo Science 2015; 64: 633-44. Robert P Friedland. Fish consumption and the risk of Alzheimer's disease: is it time to make dietary recommendations? Archives of Neurology 2003; 60: 923-4. Okubo, T. et al., PC-DHA (DHA-bound phosphatidylcholine) affects REM sleep amount - The influence of dietary environment on sleep -, Journal of the Japanese Society of Sleep Environment, 8, 9-14 (2011). Ikeda, I. Fundamentals and Applications of Functional Lipids in Foods. CMC Publishing; 2018. pp.119-123. N Sinn, et al., Effects of n-3 fatty acids, EPA vs. DHA, on depressive Symptoms, quality of life, memory and executive function in older adults with mild cognitive impairment: a 6-month randomized controlled trial. British Journal of Nutrition 2012 Jun;107:1682-93.

[0006] The purpose of the present disclosure is to provide new means for improving cognitive function.

[0007] The present disclosure provides a composition for improving cognitive function comprising a fish roe lipid preparation.

[0008] The present disclosure provides new means for improving cognitive function.

[0009] The study flow chart shows the process from the allocation of test foods to the analysis.

[0010] In this disclosure, when a numerical value is accompanied by the term "about," it is intended to encompass a range of ±10% of that value. For example, "about 20" is intended to include "18 to 22." A range of numerical values ​​includes all values ​​between and at the endpoints. When "about" refers to a range, it applies to both endpoints of the range. Thus, for example, "about 20 to 30" is intended to include "18 to 33." When percentages of ingredients are expressed in this disclosure, they are by weight unless otherwise specified.

[0011] Unless otherwise specified, terms used in this disclosure have the meanings commonly understood by those skilled in the art of organic chemistry, medicine, pharmacology, molecular biology, microbiology, etc. Below, definitions of some terms used in this disclosure are provided, but these definitions take precedence over common understandings in this disclosure.

[0012] (Method for producing fish roe lipid preparation) The fish roe lipid preparation is prepared, for example, from fish roe of the Salmonidae family. The Salmonidae family includes the Pacific Salmon, Atlantic Salmon, Salvelinus, and Sakhalin, with the Pacific Salmon or Atlantic Salmon being more preferred. Examples of Pacific salmon include chum salmon (Oncorhynchus keta), coho salmon (Oncorhynchus kisutch), pink salmon (Oncorhynchus gorbuscha), cherry salmon (Oncorhynchus masou masou), yamame (Oncorhynchus masou masou), Taiwan salmon (Oncorhynchus masou formosanus), satsukimasu (Oncorhynchus masou ishikawae), amago (Oncorhynchus masou ishikawae), Biwa salmon (Oncorhynchus masou rhodurus), rainbow trout (Oncorhynchus mykiss), Chinook salmon (Oncorhynchus tshawytscha), sockeye salmon (Oncorhynchus nerka), kokanee (Oncorhynchus nerka), and kunimasu (Oncorhynchus kawamurae). Examples of Atlantic salmon include Atlantic salmon (Salmo salar) and brown trout (Salmo trutta). In one embodiment, the fish roe lipid preparation is prepared from pink salmon (Oncorhynchus gorbuscha) roe.

[0013] In the present disclosure, the term "fish roe" refers to any level of processing, unless otherwise specified. Therefore, in the present disclosure, the term "fish roe lipid preparation" also includes lipid preparations made from fish roe extract, fish roe oil, purified fish roe oil, or any dried product thereof. In a particularly preferred embodiment, the fish roe lipid preparation is prepared from salmon roe, salmon roe, or a processed product thereof, such as salmon roe extract, salmon roe extract, salmon roe oil, or purified salmon roe oil.

[0014] Specifically, extraction of a fish roe lipid preparation from raw materials, such as fish roe or processed products thereof, can be carried out by mixing the raw materials with a low-polarity organic solvent. Examples of low-polarity solvents include, but are not limited to, one or more organic solvents selected from the group consisting of 60-99% aqueous ethanol, ethanol, hexane, isopropyl alcohol, ethyl acetate, acetone, ether, chloroform, and methanol. The extraction temperature using an organic solvent is 0-90°C, preferably 30-70°C. It is known that the phospholipid content of the resulting extract can be increased by manipulating the water content of aqueous ethanol (see Oleoscience, Vol. 2, No. 2, pp. 67-74). Extraction of a fish roe lipid preparation from raw materials, such as fish roe or processed products thereof, can also be carried out by supercritical fluid extraction using carbon dioxide. For example, the fish roe lipid preparation can be that described in WO 2021 / 132516, particularly Salmon Roe Oil PL40.

[0015] (Phospholipids) In one embodiment, the phospholipid content of the fish roe lipid preparation is increased. Phospholipids refer to lipids having phosphorus in the form of phosphate esters, and include glycerophospholipids and sphingophospholipids. Representative glycerophospholipids include phosphatidylcholine (PC), α-glycerophosphocholine (α-GPC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), 1-lysophosphatidylcholine (LPC-1), 2-lysophosphatidylcholine (LPC-2), and 2-lysophosphatidylethanolamine (LPE-2). Representative sphingophospholipids include sphingomyelin (SM) and dihydrosphingomyelin (DHSM). The phospholipid content of the fish roe lipid preparation is, for example, about 26% or more, and may be about 30% or more, preferably about 35% or more, more preferably about 37.5% or more, and even more preferably about 40% or more. The upper limit of the phospholipid content in the fish roe lipid preparation is not particularly limited, but as the phospholipid content increases, the viscosity increases, and if the viscosity exceeds a certain level, the preparation becomes difficult to handle during production, so the upper limit is, for example, about 50% or less.

[0016] In one embodiment, the fish roe lipid preparation comprises at least one selected from the group consisting of α-GPC, SM, and DHSM, and in a preferred embodiment, it comprises SM and DHSM. The contents of α-GPC, SM, and DHSM are not particularly limited. The content of α-GPC in the fish roe lipid preparation is, for example, about 0.050% to 0.60%. The content of SM in the fish roe lipid preparation is, for example, about 0.8% to 2.3%. The content of DHSM in the fish roe lipid preparation is, for example, about 0.050% to 0.40%.

[0017] The phospholipids other than α-GPC, SM, and DHSM contained in the fish roe lipid preparation are not particularly limited, and preferably contain a large amount of PC, PE, PI, LPC-2, etc. The PC content in the fish roe lipid preparation is, for example, about 24 to 45%, preferably about 26 to 43%, more preferably about 28 to 41%, and even more preferably about 30 to 39%. The PE content in the fish roe lipid preparation is, for example, about 0.90 to 2.3%. The PI content in the fish roe lipid preparation is, for example, about 0.80 to 1.8%. The LPC-2 content in the fish roe lipid preparation is, for example, about 0.60 to 3.0%.

[0018] In the fish roe lipid preparation, PC is contained in a relatively large amount in the phospholipids. The PC content in the phospholipids is, for example, about 74% or more, preferably about 80% or more. In a fish roe lipid preparation with an increased phospholipid content, the PC content in the phospholipids is even higher. Therefore, in a preferred embodiment, the phospholipid content in the fish roe lipid preparation is about 35% or more, more preferably about 40% or more, and in this case, the PC content in the phospholipids is about 74% or more, preferably about 80% or more.

[0019] (Fatty Acid Composition) The composition ratio of DHA in the constituent fatty acids of lipids in fish roe lipid preparations is relatively high. Fish roe is known to contain DHA in the form of phospholipids or triglycerides (TG). Specifically, the composition ratio of DHA in the constituent fatty acids of lipids in fish roe lipid preparations is, for example, about 15% or more, preferably about 18% or more, more preferably about 22% or more, and even more preferably about 24% or more. The upper limit of the composition ratio of DHA in the constituent fatty acids of lipids in fish roe lipid preparations is not particularly limited, but is, for example, about 46% or less, preferably about 40% or less, more preferably about 35% or less, and even more preferably about 30% or less. In the present disclosure, when the composition ratio of a specific fatty acid in the constituent fatty acids of lipids is expressed as a percentage, it is based on the area of ​​a chart obtained by analyzing the fatty acid composition by gas chromatography, unless otherwise specified.

[0020] The composition ratio of EPA in the constituent fatty acids of the lipids in the fish roe lipid preparation is relatively low, for example, about 25% or less, preferably about 23% or less, more preferably about 21% or less, and even more preferably about 19% or less. The lower limit of the composition ratio of EPA in the constituent fatty acids of the lipids in the fish roe lipid preparation is not particularly limited, but may be, for example, about 5.0% or more, or may be about 6.0% or more, preferably about 8.0% or more, more preferably about 10% or more, and even more preferably about 11% or more or about 12% or more.

[0021] The fish roe lipid preparation contains, as constituent fatty acids, myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1, n9c), eicosenoic acid (C20:1), and docosapentaenoic acid (DPA) (C22:5). Other constituent fatty acids include myristoleic acid (C14:1), pentadecenoic acid (C15:1), heptadecenoic acid (C17:1), docosenoic acid (C22:1), tetracosenoic acid (C24:1), linoleic acid (C18:2n-6), α-linolenic acid (C18:3n-3), γ-linolenic acid (C18:3n-6), eicosadienoic acid (C20:2n-6), eicosatrienoic acid (C20:3n-6), arachidonic acid (C20:4n-6), and docosadienoic acid (C22:2).

[0022] Fish egg lipid preparation contains a large amount of DHA-bound phospholipids.In detail, the weight of DHA per 100g of fish egg lipid preparation is, for example, about 10g or more, preferably about 12g or more, more preferably about 14g or more, and even more preferably about 15g or more.The upper limit of the weight of DHA per 100g of fish egg lipid preparation is not particularly limited, but is, for example, about 30g or less, preferably about 25g or less, more preferably about 22g or less, and even more preferably about 20g or less.

[0023] The weight of EPA per 100 g of the fish roe lipid preparation is, for example, about 5.0 g or more, more preferably about 6.0 g or more. The upper limit of the weight of EPA per 100 g of the fish roe lipid preparation is not particularly limited, but is, for example, about 20 g or less, preferably about 15 g or less.

[0024] One of the DHA-bound phospholipids contained in fish egg lipid preparations is palmitoyldocosahexaenoylphosphatidylcholine (PDPC), a phosphatidylcholine (38:6) in which one of the C-1 and C-2 acyl groups is hexadecanoyl (16:0) and the other is docosahexaenoyl (22:6).

[0025] Another DHA-bound phospholipid contained in fish egg lipid preparations is stearoyl docosahexaenoyl phosphatidylcholine (SDPC), which is a phosphatidylcholine (40:6) in which one of the C-1 and C-2 acyl groups is stearoyl (18:0) and the other is docosahexaenoyl (22:6).

[0026] One of the phospholipids contained in fish egg lipid preparations is an ether-type phospholipid. An ether-type phospholipid is a phospholipid having a hydrocarbon chain formed by an ether bond. In particular, a glycerophospholipid having a hydrocarbon chain formed by a vinyl ether bond at the sn-1 position and a fatty acid bonded at the sn-2 position is called a plasmalogen.

[0027] In one embodiment, the fish roe lipid preparation comprises any one selected from the group consisting of DHA-bound ether-type phospholipids and EPA-bound ether-type phospholipids.

[0028] (Components other than lipids) The fish roe lipid preparation may contain components other than lipids. Components other than lipids include inorganic substances such as proteins, sodium, potassium, and phosphorus. Lipids refer to biologically derived substances that are soluble in non-polar solvents, including simple lipids, complex lipids, and derived lipids (fatty acids, terpenoids, steroids, carotenoids, etc.). The content of components other than lipids in the fish roe lipid preparation is, for example, 10.0% or less, preferably 8.0% or less, more preferably 7.5% or less, and even more preferably 7.0% or less.

[0029] The fish roe lipid preparation may contain astaxanthin. The content thereof in 100 g of the fish roe lipid preparation is, for example, about 0.7 mg or more, preferably about 1.0 mg or more, more preferably about 1.2 mg or more, and even more preferably about 1.5 mg or more. The upper limit of the astaxanthin content in 100 g of the fish roe lipid preparation is not particularly limited, but is, for example, about 23 mg or less, preferably about 20 mg or less, more preferably about 10 mg or less, and even more preferably about 5.0 mg or less.

[0030] (Uses) As shown in the examples below, the composition of the present disclosure improves cognitive function. In this disclosure, improving cognitive function includes maintaining the state, inhibiting decline, maintaining an improved state, and inhibiting decline from an improved state. As used in this disclosure, "cognitive function" includes language ability in a subject who has a regular exercise habit. Exercise refers to physical activity that is planned, intentionally, and continuously performed for the purpose of maintaining or improving physical strength. For example, exercise includes strength training, aerobic exercise, sports, walking, and active hobbies, specifically stretching, gymnastics, yoga, mountain climbing, hiking, dance, golf, gateball, and swimming (including water walking). In one embodiment, the composition of the present disclosure improves language ability in a subject who has a regular exercise habit. A regular exercise habit means exercising regularly, for example, daily, at least three times a week, at least twice a week, at least once a week, at least three times a month, or at least twice a month. In one embodiment, regular exercise habits refer to exercise such as stretching, gymnastics, yoga, mountain climbing, hiking, dancing, golf, gateball, or swimming (including underwater walking) performed at least once a week. Language ability refers to the ability to understand spoken language and written text, and to communicate using language. In one aspect, it refers to the ability to understand the meaning of a sentence, remember it, and accurately verbalize it, and can be assessed, for example, by the repetition score of the Montreal Cognitive Assessment (MoCA). MoCA is available in various languages, including the Japanese version (MoCA-J).

[0031] Compositions containing fish roe lipid preparations also improve mental health or mental quality of life (QOL) as assessed by the SF-36. In the present disclosure, improvement of mental health or mental quality of life (QOL) includes maintaining the state, preventing decline, maintaining an improved state, and preventing decline from an improved state. The SF-36 is a scale for measuring health-related quality of life (HRQOL). For example, in Japan, the SF-36v2, an improved version of the original SF-36 (Japanese version is version 1.2), is used. The SF-36 consists of 36 items, and NBS scores (scores based on a national standard score system) for eight health concepts (physical function, role function (physical), bodily pain, general health, vitality, social function, role function (mental), and mental health) can be calculated, along with component summary scores. A higher score indicates a higher QOL.

[0032] A high score on "Mental Health" on the SF-36 indicates a calm, happy, and peaceful mood over the past month. In some embodiments, improving mental health is an improvement in at least one of nervousness, depression, and gloomy mood. In some embodiments, improving mental health is an improvement to a calm or happy mood.

[0033] Examples of component summary scores include a 3-component summary score (3MCS; physical aspect, mental aspect, social aspect) based on factor coefficients from a 2002 Japanese survey, a 2-component summary score (2MCS; physical aspect, mental aspect) based on factor coefficients from a 1995 U.S. national survey, and a 2-component summary score (2MCS (Japanese version); physical aspect, mental aspect) based on factor coefficients from a 1995 Japanese national survey. In one embodiment, mental aspect QOL is assessed using the 3MCS or the 2MCS (Japanese version).

[0034] The subject is typically a human. The subject may or may not have a cognitive impairment. Cognitive impairment includes forgetfulness and mild cognitive impairment. A forgetful subject includes a subject who has subjective symptoms of forgetfulness or a subject whose close relative has pointed out symptoms of forgetfulness. The subject's age is not limited, but may be middle-aged or elderly, specifically, 45 years or older, 50 years or older, 55 years or older, 60 years or older, or 65 years or older, and may be 95 years or younger, 85 years or younger, 80 years or younger, or 75 years or younger.

[0035] The composition of the present disclosure may be a pharmaceutical composition. The method of administration of the pharmaceutical composition is not particularly limited, and is preferably oral administration, transdermal administration, or nasal administration, more preferably oral administration. Dosage forms for oral administration include granules, fine granules, powders, coated tablets, tablets, powders, soft capsules, hard capsules, microcapsules, chewable tablets, liquids, suspensions, emulsions, etc. Dosage forms for transdermal administration include patches, tapes, sprays, lotions, creams, ointments, liquids, emulsions, suspensions, etc. Dosage forms for nasal administration include nasal drops, nasal sprays, etc.

[0036] These dosage forms are produced by formulating the formulations using conventional methods. Furthermore, various pharmaceutically acceptable pharmaceutical substances can be blended as required for the formulation. The pharmaceutical substances can be appropriately selected depending on the dosage form of the formulation, and examples thereof include buffering agents, surfactants, stabilizers, preservatives, excipients, diluents, additives, disintegrants, binders, coating agents, lubricants, flavoring agents, sweeteners, solubilizers, etc.

[0037] The composition of the present disclosure may also be a food composition. The food composition may be in the form of a typical processed food. For example, the food composition may be a solid food, or a liquid food such as a beverage, a drinkable supplement, a powdered beverage, or a soup. Specifically, the food composition may be consumed as, for example, juice, confectionery, jelly, tablet, dressing, seasoning, or the like.

[0038] Such foods may also be provided as health functional foods or dietary supplements. Health functional foods include, for example, foods for specified health uses, foods with nutrient functions, and foods with functional claims. Health functional foods can be labeled as being used for purposes such as improving cognitive function. The labeling may be directly displayed on the packaging, container, label, tag, attached documents, etc., accompanying the product, or indirectly displayed through advertising and promotional activities. Dietary supplements include, for example, nutritional supplements and health supplements.

[0039] The intake amount of the composition of the present disclosure can be about 100 mg to 10,000 mg of the fish roe lipid preparation per day, preferably about 300 mg to 5,000 mg, more preferably about 500 mg to 2,500 mg, even more preferably about 700 mg to 1,500 mg, for example, about 1,000 mg.

[0040] Alternatively, the amount of intake of the composition of the present disclosure can be an amount that results in about 20 mg or more of DHA being ingested per day, preferably about 50 mg or more, more preferably about 100 mg or more, and even more preferably about 150 mg or more. Alternatively, the amount of intake of the composition of the present disclosure can be an amount that results in about 2000 mg or less of DHA being ingested per day, preferably about 1000 mg or less, more preferably about 750 mg or less, even more preferably about 500 mg or less, even more preferably 300 mg or less or less than 300 mg, and especially preferably about 200 mg or less. In some embodiments, the amount of intake of the composition of the present disclosure is an amount that results in about 150 mg (e.g., 135 mg to 165 mg) or about 160 mg (e.g., 144 mg to 176 mg) of DHA being ingested per day.

[0041] The amount of intake of the composition of the present disclosure can be an amount that results in an intake of about 10 mg or more of EPA per day, preferably about 30 mg or more, more preferably about 50 mg or more, and even more preferably about 60 mg or more. The amount of intake of the composition of the present disclosure can be an amount that results in an intake of about 500 mg or less of EPA per day, preferably about 300 mg or less, and more preferably about 150 mg or less. In some embodiments, the amount of intake of the composition of the present disclosure is an amount that results in an intake of about 60 mg (e.g., 54 mg to 66 mg), about 100 mg (e.g., 90 mg to 110 mg), or about 110 mg (e.g., 99 mg to 121 mg) of EPA per day.

[0042] Each component of the daily intake amount may be contained in one composition, or may be dispersed among multiple compositions to be taken in one day. For example, each component of the daily intake amount of the composition may be dispersed among 2 to 15, preferably 3 to 10, e.g., 4 capsules to be taken in one day.

[0043] The composition of the present disclosure may be taken once or multiple times. When taken multiple times, for example, once to several times a day, for example, once, twice, or three times a day, daily or every few days, for example, every 1, 2, 3, or 7 days. The duration of intake is not limited, and the composition may be taken continuously for, for example, one week or more, preferably one month or more, more preferably two months or more, and particularly preferably three months or more, until cognitive function, mental health, or mental aspects of QOL improve. There may be a period during which the intake is discontinued. In one embodiment, the composition of the present disclosure is taken daily for at least 12 weeks.

[0044] The composition of the present disclosure can be used alone or in combination with one or more additional ingredients. "Combined use" of ingredients means not only the use of a dosage form containing all ingredients or the use of a combination of dosage forms containing each ingredient separately, but also the simultaneous intake of each ingredient or delayed intake of any ingredient, as long as they are used for the same purpose. Two or more additional ingredients can also be used in combination. For example, a composition containing one or more additional ingredients in addition to a fish roe oil preparation can be used.

[0045] An example of a suitable component for use in combination is astaxanthin. The daily intake of astaxanthin can be about 50 μg or more, preferably about 100 μg or more, more preferably about 200 μg or more, and about 1000 μg or less, preferably about 400 μg or less, for example, about 200 μg (e.g., 180 μg to 220 μg). While astaxanthin may be contained in fish roe lipid compositions, astaxanthin from other sources may also be used. For example, astaxanthin extracted from natural sources such as krill, salmon, trout, Adonis asiaticus, red yeast, and Haematococcus algae, or synthetic products may be used. Preferably, the astaxanthin is contained in the pigment of Haematococcus algae. The extraction solvent used to obtain astaxanthin from natural sources may be either an aqueous solvent or an organic solvent. Examples of organic solvents that can be used include methanol, ethanol, isopropanol, acetone, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, ethyl acetate, ether, and hexane. Supercritical carbon dioxide can also be used. These solvents can be used alone or in combination. In one embodiment, astaxanthin protects and stabilizes DHA and EPA from peroxidation.

[0046] Specific examples of Haematococcus algae include Haematococcus pluvialis, Haematococcus lacustris, Haematococcus capensis, Haematococcus droebakensis, Haematococcus zimbabwiensis, etc. Commercially available Haematococcus algae extracts can also be used, such as ASTOTS-S, ASTOTS-5O, and ASTOTS-10O manufactured by Fuji Film Corporation, Astareal Oil 50F and Astareal Oil 5F manufactured by Fuji Chemical Industry Co., Ltd., Astaxanthin-5C and Astaxanthin-20C manufactured by Oryza Oil & Fat Chemical Co., Ltd., Astaxanthin 5% Oil manufactured by Bioactives Japan, and Astabio manufactured by Biogenic Co., Ltd. （登録商標） Examples of astaxanthin derived from krill include Astax-S manufactured by Marine Daio Co., Ltd.

[0047] An example of a suitable ingredient for use in combination is lecithin. The daily intake of lecithin can be about 50 mg or more, preferably about 60 mg or more, more preferably about 70 mg or more, and can be about 500 mg or less, preferably about 300 mg or less, more preferably about 150 mg or less, for example, about 100 mg (e.g., 90 mg to 110 mg). Lecithin is a general term for lipid products containing phospholipids, and can include phospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, and phosphatidic acid. Examples of lecithin include soybean lecithin, enzymatically hydrolyzed soybean lecithin, hydrogenated soybean lecithin, egg yolk lecithin, hydrogenated phospholipids, milk-derived phospholipids, lysolecithin, phosphatidylcholine, and phosphatidylserine, and preferably plant lecithin, particularly soybean lecithin. Soybean lecithin can be produced as a by-product of the degumming process during the production of soybean oil. It may be further decolorized and purified. Commercially available lecithin can also be used, such as purified soybean lecithin manufactured by Nisshin Oillio Co., Ltd., purified egg yolk lecithin manufactured by Asahi Kasei Pharma Corporation, and egg yolk lecithin PL-100M manufactured by Kewpie Corporation.

[0048] When the composition of the present disclosure contains lecithin, in one embodiment, the content of PC contained in the composition is, for example, about 24-45%, preferably about 26-43%, more preferably about 28-41%, and even more preferably about 30-39%. The content of PE contained in the composition is, for example, about 0.90-3.5%. The content of PI contained in the composition is, for example, about 0.80-2.5%. The content of LPC-2 contained in the composition is, for example, about 0.60-3.0%. The content of SM contained in the composition is, for example, about 0.8%-2.3%. The content of phosphatidic acid contained in the composition is, for example, about 0.10-1.0%. The content of DHSM contained in the composition is, for example, about 0.050%-0.40%. The phospholipid content in this composition is about 35% or more, more preferably about 40% or more, and in this case, the PC content in the phospholipid is about 74% or more, preferably about 77% or more.

[0049] In one embodiment, a cognitive function improving agent comprising a fish roe lipid preparation is provided. In one embodiment, a method for improving cognitive function is provided, comprising ingesting a fish roe lipid preparation to a subject in need of cognitive function improvement. In one embodiment, a fish roe lipid preparation for improving cognitive function is provided. In one embodiment, a use of a fish roe lipid preparation for improving cognitive function is provided. In one embodiment, a use of a fish roe lipid preparation in the manufacture of a composition for improving cognitive function is provided.

[0050] In one aspect, an agent for improving mental health or mental aspects of QOL is provided, comprising a fish roe lipid preparation. In one aspect, a method for maintaining, enhancing, and / or improving mental health or mental aspects of QOL is provided, comprising administering a fish roe lipid preparation to a subject in need of such improvement. In one aspect, a fish roe lipid preparation for improving mental health or mental aspects of QOL is provided. In one aspect, a use of a fish roe lipid preparation for improving mental health or mental aspects of QOL is provided. In one aspect, a use of a fish roe lipid preparation in the manufacture of a composition for improving mental health or mental aspects of QOL is provided.

[0051] For example, the following embodiments are provided. [1] A composition for improving cognitive function, comprising a fish roe lipid preparation. [2] The composition according to item 1, wherein the cognitive function is language ability in a subject who engages in regular exercise. [3] A composition for improving cognitive function, comprising a fish roe lipid preparation that has been formulated to provide a daily intake of at least about 100 mg but less than 300 mg (preferably about 100 mg to 200 mg, more preferably about 150 mg to 200 mg) of DHA, wherein the cognitive function is language ability in a subject who engages in regular exercise. [4] The composition according to any one of items 1 to 3, further for improving mental health or mental aspects of QOL. [5] A composition for improving mental health or mental aspects of QOL, comprising a fish roe lipid preparation. [6] The composition according to item 4 or 5, wherein the composition for improving mental health. [7] The composition according to any one of items 4 to 6, wherein improving mental health is improving at least one of nervousness, depression, and gloomy mood. [8] The composition according to any one of items 4 to 6, wherein improving mental health means improving to a calm or happy mood. [9] The composition according to item 4 or 5, for improving mental QOL assessed by SF-36 2MCS (Japanese version).

[10] The composition according to item 4 or 5, for improving mental QOL assessed by SF-36 3MCS.

[11] The composition according to any one of items 1 to 10, comprising a fish roe lipid preparation containing about 30% or more (preferably about 40% or more) of phospholipids.

[12] The composition according to any one of items 1 to 11, wherein the fish roe lipid preparation contains DHA-bound phospholipids, and the composition ratio of DHA to the constituent fatty acids of the lipid is about 15% or more.

[13] The composition according to any one of items 1 to 12, wherein the fish roe lipid preparation comprises at least one selected from the group consisting of α-GPC, SM, and DHSM (preferably the group consisting of SM and DHSM).

[14] The composition according to any one of items 11 to 13, wherein the phospholipids contain about 74% or more of PC.

[15] The composition according to any one of items 1 to 14, wherein the fish roe lipid preparation contains a DHA composition ratio of about 15% or more and an EPA composition ratio of about 5 to 25% of the constituent fatty acids of the lipids.

[16] The composition according to any one of items 1 to 15, further comprising astaxanthin.

[17] The composition according to item 16, wherein the astaxanthin is derived from a pigment of Haematococcus algae.

[18] The composition according to any one of items 1 to 17, further comprising lecithin.

[19] The composition according to item 18, wherein the lecithin is soybean lecithin.

[20] The composition according to any one of items 1 to 19, which is formulated to provide a daily intake of about 100 mg or more but less than 300 mg (preferably about 100 mg to 200 mg, more preferably about 150 mg to 200 mg) of DHA.

[21] The composition according to any one of items 1 to 20, which is formulated to provide a daily intake of about 30 mg to 300 mg (preferably about 60 mg to 300 mg, more preferably about 60 mg to 150 mg) of eicosapentaenoic acid (EPA).

[22] The composition according to any one of items 1 to 21, which is formulated to provide a daily intake of about 160 mg (or 144 mg to 176 mg) of DHA.

[23] The composition according to any one of items 1 to 22, which is formulated to provide a daily intake of about 100 mg to less than 300 mg (preferably about 100 mg to 200 mg, more preferably about 150 mg to 200 mg) of DHA, about 30 mg to 300 mg (preferably about 60 mg to 300 mg, more preferably about 60 mg to 150 mg) of eicosapentaenoic acid (EPA), and about 50 mg to 500 mg (preferably about 60 mg to 300 mg, more preferably about 70 mg to 150 mg) of soybean lecithin.

[24] The composition according to item 23, further comprising about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg) of astaxanthin.

[25] The composition according to item 23, which is formulated to provide a daily intake of about 160 mg of DHA (or 144 mg to 176 mg), about 100 to 110 mg (preferably about 100 mg (or 90 to 110 mg) or about 110 mg (or 99 mg to 121 mg)) of EPA, and about 100 mg (or 90 mg to 110 mg) of soybean lecithin.

[26] The composition according to item 23, which is formulated to provide a daily intake of about 160 mg of DHA (or 144 mg to 176 mg), about 110 mg of EPA (or 99 mg to 121 mg), and about 100 mg of soybean lecithin (or 90 mg to 110 mg).

[27] The composition according to item 25 or 26, which is further formulated to provide a daily intake of about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[28] The composition according to item 23, which is formulated to provide a daily intake of about 150 mg (or 135 mg to 165 mg) of DHA, about 60 mg (or 54 mg to 66 mg) of EPA, and about 100 mg (or 90 mg to 110 mg) of soybean lecithin.

[29] The composition according to item 28, further adjusted to contain about 200 μg (or 180 μg to 220 μg) of astaxanthin for ingestion.

[30] The composition according to any of items 1 to 29, wherein the fish roe lipid preparation is derived from pink salmon.

[31] The composition according to any of items 1 to 30, which is ingested daily for at least 12 weeks.

[32] The composition according to any of items 1 to 31, wherein the subject to be ingested is middle-aged or elderly.

[33] The composition according to item 32, wherein the subject is 60 to 85 years old.

[34] The composition according to any of items 1 to 33, which is a pharmaceutical composition.

[35] The composition according to any of items 1 to 33, which is a food composition.

[36] An agent for improving cognitive function, comprising a fish roe lipid preparation.

[37] A method for improving cognitive function, comprising administering a fish roe lipid preparation to a subject in need of such improvement.

[38] A fish roe lipid preparation for improving cognitive function.

[39] Use of a fish roe lipid preparation for improving cognitive function.

[40] Use of a fish roe lipid preparation in the manufacture of a composition for improving cognitive function.

[41] An agent for improving mental health or mental aspects of QOL, comprising the fish roe lipid preparation.

[42] A method for improving mental health or mental aspects of QOL, comprising having a subject in need of improvement in mental health or mental aspects of QOL ingest the fish roe lipid preparation.

[43] A fish roe lipid preparation for improving mental health or mental aspects of QOL.

[44] Use of a fish roe lipid preparation for improving mental health or mental aspects of QOL.

[45] Use of a fish roe lipid preparation in the manufacture of a composition for improving mental health or mental aspects of QOL.

[0052] All documents cited in this specification are incorporated herein by reference. The present invention will be described in more detail in the following examples, but the present invention is not limited to these examples. The above descriptions are all non-limiting, and the present invention is defined in the appended claims, and various modifications are possible within the scope of the technical idea thereof.

[0053] <Production of fish roe lipid preparation from salmon roe> A fish roe lipid preparation was produced by the method described in WO2021 / 132516. Specifically, frozen and crushed raw salmon roe (Oncorhynchus gorbuscha) (15 kg) was extracted with aqueous ethanol (150 L), and the residue was filtered off. The residue was returned to the extractor, and ethanol (75 L) was added again for extraction. The solvent was removed from the obtained filtrate under reduced pressure, and a composition (40 PL) containing approximately 40% phospholipids in a red oily substance was produced (Tables 1 and 2).

[0054] *Quantitative values ​​are the amounts of constituent fatty acids in total lipids.

[0055] <Production of Soft Capsules> Haematococcus algae pigment (astaxanthin) and soybean lecithin (vegetable lecithin) were added to the 40PL to produce soft capsules "ONO-SR / AST-SOYPC" containing the ingredients in the table below per capsule (Table 3). The amounts of phospholipid classes contained in the soft capsules are shown in Table 4. *The raw material gelatin is derived from cows

[0056]

[0057] The specifications of ONO-SR / AST-SOYPC are as shown in the table below.

[0058] <Clinical trial> I. Subjects and methods 1. Subjects The subjects were 100 men and women aged between 60 and 85 who either had subjective symptoms of forgetfulness or had a close relative who had been told they had symptoms of forgetfulness (including those diagnosed with MCI). Prior to the study, a consent form was issued to the subjects, the purpose and content of the study were fully explained, and written consent was obtained from the subjects based on their own free will. Based on the results of the pre-tests of subjects who were able to give consent, subjects who did not fall under the following exclusion criteria were selected.

[0059] (1) Individuals who regularly consume health foods rich in DHA, EPA, astaxanthin, or soy lecithin. (2) Individuals who continuously consume, at least once a week, pharmaceuticals, health foods, foods for specified health uses, functional foods, or foods with functional claims that contain ingredients with cognitive function improving, antioxidant, or blood flow improving effects, which may affect the results of the study. (3) Individuals who have difficulty performing cognitive function tests due to poor eyesight, or who have been diagnosed with amblyopia or blindness. (4) Individuals who have difficulty performing cognitive function tests due to poor hearing, or who have been diagnosed with hearing loss or deafness. (5) Individuals with a history or current illness of mental disorders, dementia, neurological disorders, or cerebrovascular diseases. (6) Individuals with current menopausal disorders. (7) Individuals who are taking pharmaceuticals that may affect the results of the study (hyperlipidemic drugs, antipsychotics, antianxiety drugs, antidepressants / manic drugs, antiparkinsonian drugs, antiepileptic drugs, anticoagulants, etc.). (8) Persons with a history of serious illnesses that required medication (malignant tumors, diabetes, liver disease (hepatitis), kidney disease, heart disease, etc.). (9) Persons with a past or current history of drug or alcohol dependence. (10) Persons suspected of having dementia based on the cognitive function test in the pre-test or a medical interview with a doctor. (11) Persons whose lifestyles become irregular multiple times during the study period due to night shifts, etc. (12) Persons who currently smoke or have quit smoking within 12 months prior to the pre-test. (13) Persons who have undergone a cognitive function test (Mini-Mental State Examination (MMSE)) or the Japanese version of the Montreal Cognitive Assessment (MoCA-J)) within 12 months prior to the pre-test. (14) Persons who have donated 200 mL or 400 mL of whole blood or received a blood transfusion within 3 months prior to the pre-test. (15) Persons who have participated in another clinical study within 1 month prior to the pre-test, or who plan to participate during the study period. (16) Anyone who is deemed unsuitable as a subject based on the clinical test values ​​and measurements of the pre-examination. (17) Anyone who is at risk of developing an allergy related to the study. (18) Anyone who is deemed unsuitable as a subject based on the responses to the lifestyle questionnaire. (19) Anyone who is otherwise deemed unsuitable as a subject by the responsible physician.

[0060] Prior to the study, the researcher instructed all subjects not to violate the following rules. Throughout the study, (1) make as few changes as possible to their lifestyle habits, including drinking, eating, sleeping, and work, as they had before participating in the study. (2) Limit excessive exercise that deviates significantly from their usual range, as well as dieting and overeating. (3) Do not start a new exercise routine or discontinue an existing exercise habit. (4) Do not travel or go on business trips abroad. (5) Do not consume any new health foods, foods for specified health uses, foods with nutrient functions, or foods with functional claims. Continue to consume health foods, etc. that you are currently consuming during the study period. (6) Do not consume any medicines, health foods, foods for specified health uses, foods with functional claims, or energy drinks that contain ingredients that may affect cognitive function, antioxidants, or improving blood flow. (7) Do not donate blood. (8) Consume the specified amount of study food every day and keep a daily diary. During the preliminary (blood sampling) test, (9) Refrain from drinking alcohol from the morning of the day before the test until the end of the test on the day. (10) Do not engage in excessive exercise from the day before the test until the end of the test on the day of the test. (11) On the day of the test, fast for at least 8 hours after waking up until the end of the test (water may be consumed). For the pre-test (cognitive) test: (12) Get sufficient sleep and rest the day before the test. (13) Do not engage in any activities that may have a significant impact on cognitive function immediately before the test. (14) Observe the same rules as items (9) and (10). For the 6-week test and 12-week (cognitive) test: (15) Observe the same rules as items (9), (10), (12), and (13). (16) On the day of the test, come to the hospital without consuming any study foods. (17) As a general rule, cognitive function testing should start at approximately the same time as the pre-test (cognitive) test. The test should be conducted in the morning or afternoon, and the start time of the pre-test (cognitive) cognitive function test should be within a ±2-hour range. (18) Live your life so that your work schedule before the pre-test (cognitive) and the day of the test do not change drastically. (19) Live your life so that your exercise schedule the day before and the day of the test and your last meal before the test do not change drastically. At the time of the 12th week (blood sampling) test, (20) observe the same contents as items (9) to (12) and (16).

[0061] 2. Study Foods There were two types of study foods: test food and placebo. The test food contained 1000 mg of salmon roe oil (containing 150 mg of DHA and 60 mg of EPA) in four capsules, the daily intake amount. Other ingredients included astaxanthin, soy lecithin, gelatin, glycerin, water, and caramel coloring in soft capsules. The placebo was a soft capsule containing safflower oil instead of salmon roe oil. Each study food was packaged in a bag containing four capsules, the daily intake amount, so that the test food and placebo could not be distinguished from each other by the packaging.

[0062] 3. Study design This study was a randomized, double-blind, placebo-controlled, parallel-group comparative study with an intake period of 12 weeks. The allocation manager, who was not directly involved in the study, created an allocation list using a random number table and assigned allocation numbers to the study foods based on this. The allocation list was sealed by the allocation manager and kept sealed until it was opened. The principal investigator, subjects, medical facility staff, and all other staff involved in this study were kept blind.

[0063] Before the intervention, all subjects in both groups underwent a pre-examination, including anthropometric measurements (height, weight, and BMI calculation), physical examinations (blood pressure, pulse rate), various clinical tests (hematology, blood chemistry, and urinalysis), cognitive function tests (MMSE, MoCA-J), and the MOS 36-Item Short-Form Health Survey (SF-36) (S Fukuhara, et al. Translation, adaptation, and validation of the SF-36 Health Survey for use in Japan. J Clin Epidemiol 1998; 51: 1037-44; S Fukuhara, et al. Psychometric and clinical tests of validity of the Japanese SF-36 Health Survey. J Clin Epidemiol 1998; 51: 1045-53). The obtained values ​​were used as baseline values.

[0064] Each subject was randomly assigned to take one bag of the study food (four tablets) per day after meals with water or lukewarm water, without chewing, throughout the 12-week intervention period. During the intervention period, subjects were asked to record their intake of the study food, changes in their physical condition, and medication use in a diary. Six and 12 weeks after the start of intake, all subjects visited the hospital for anthropometric measurements, physical examinations, and cognitive function tests. After 12 weeks, clinical tests, special tests, and the SF-36 were also conducted.

[0065] 4. Evaluation of Efficacy To evaluate the efficacy of the study food, the MoCA-J was used as the primary endpoint, and the MMSE and SF-36 were used as secondary endpoints. Both the MoCA-J and MMSE are tests that can perform multifaceted evaluations of various areas of cognitive function. In addition, since both are used to screen for MCI, they were used to evaluate the cognitive function of healthy subjects in this study, including those with MCI. Details of each test are described below.

[0066] 1) MoCA-J The MoCA-J consists of 11 items: trail making, visuospatial cognitive function (cube), visuospatial cognitive function (clock drawing), naming, memory, attention (forward and backward repetition, vigilance, and calculation), repetition, word recall, abstract thinking, delayed recall, and orientation. It was developed to screen for MCI. A score of 25 or less out of a possible 30 points indicates MCI, with reported sensitivity of 80-100% and specificity of 50-87% (Ziad S Nasreddine, et al. The Montreal Cognitive Assessment MoCA: a brief screening tool for mild cognitive impairment. J Am Geriatr Soc 2005; 53: 695-9; Bruce A Fage, et al. Mini-Cog for the diagnosis of Alzheimer's disease dementia and other dementias within a community setting. Cochran Database Syst Rev 2015: CD010860). In this study, it was noted that the attention (calculation) and orientation items overlap with those of the MMSE, and the MoCA-J and MMSE were administered simultaneously, as in a previous study (Tashiro Daisuke et al., Age-related comparison of cognitive function using the MMSE and MoCA-J in community-dwelling older adults. Physical Therapy Science 2019; 34: 331-5). The tests were administered on the same day, in the order of MMSE and MoCA-J. The attention (calculation) items in the MoCA-J and the attention and calculation items in the MMSE were the same calculation task (repeatedly subtracting 7 from 100 five times), but considering the time from memorization to delayed recall in the MoCA-J, the calculation task was also administered in the MoCA-J, and the scores from the MMSE were used. The orientation of the MoCA-J and the orientation to time and orientation to place of the MMSE were conducted only in the MMSE because the tasks of date (year, month, day, day of the week) and place (city, building) were the same, and the scores from the MMSE were used. In both cases, a higher score indicates a higher level of cognitive function.

[0067] 2) MMSE The MMSE is a test consisting of 11 items: orientation to time, orientation to place, memorization, attention and calculation, recall, naming, repetition, comprehension, reading, writing, and drawing. It has been reported that a score of 23 or less out of a possible 30 points indicates dementia (sensitivity 81%, specificity 89%), and a score of 27 or less indicates MCI (sensitivity 45-60%, specificity 65-90%) (MF Folstein, et al. "Mini-mental state". A practical method for grading the cognitive state of patients for the clinician. J Psychiatr Res 1975; 12: 189-98.; Kelvin KF Tsoi, et al. Cognitive Tests to Detect Dementia: A Systematic Review and Meta-analysis. JAMA Intern Med 2015; 175: 1450-8; Syed H Tariq, et al. Comparison of the Saint Louis University mental status examination and the mini-mental state examination for detecting dementia and mild neurocognitive disorder: a pilot study. Am J Geriatr Psychiatry 2006; 14: 900-10; Judith Saxton, et al. Computer assessment of mild cognitive impairment. Postgrad Med 2009; 121: 177-85.; Daniel I Kaufer, et al. Cognitive screening for dementia and mild cognitive impairment in assisted living: comparison of 3 tests. J Am Med Dir Assoc 2008; 9: 586-93.) In each case, higher scores indicate better cognitive function.

[0068] 3) SF-36 The Japanese version of the SF-36v2 was used to assess health-related quality of life (Health Related Quality of Life). The SF-36 consists of 36 items, and NBS scores (scores based on a scoring method based on national norms) for eight health concepts (physical function, daily role function (physical), bodily pain, general health, vitality, social function, daily role function (mental), and mental health) can be calculated. Component summary scores can also be calculated from these scores (three-component summary scores (physical, mental, and social aspects) based on factor coefficients from the 2002 Japan survey, two-component summary scores (physical and mental aspects) based on factor coefficients from the 1995 U.S. national survey, and two-component summary scores (physical and mental aspects) based on factor coefficients from the 1995 Japan national survey). A higher score indicates a higher QOL.

[0069] 5. Safety Evaluation Based on the results of the interview, physical measurements, and physical examinations at each visit, as well as entries in the diary, the principal investigator treated all undesirable or unintended injuries, illnesses, or symptoms thereof that occurred in the subjects during the intake period as adverse events. In addition, clinical tests were conducted for the following items: hematological tests (white blood cell count, red blood cell count, hemoglobin, hematocrit, platelet count), blood biochemistry tests (total protein, albumin, total bilirubin, direct bilirubin, indirect bilirubin, ALP, AST (GOT), ALT (GPT), LD, γ-GT, total cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol, urea nitrogen, creatinine, uric acid, Na, K, CL, blood glucose, HbA1c (preliminary test only)), and urinalysis (protein (qualitative), sugar (qualitative), occult blood reaction (qualitative)). The investigator-in-chief determined whether the test values ​​of individual subjects were abnormal fluctuations that constituted adverse events based on the reference values ​​established by the medical institution, and with reference to the criteria for determining abnormal fluctuations established by the Japanese Society of Chemotherapy, CTCAE v5.0-JCOG, and the Japan Society of Ningen Dock Judgment Classification (revised April 1, 2018, partially revised December 14, 2018). Furthermore, the investigator-in-chief investigated and considered the relationship with the study food, and determined whether each adverse event was a side effect of the study food.

[0070] For safety analysis, subjects who had taken the study food at least once were included for adverse events, and subjects who had completed all of the prescribed study schedules and contents for measurements and test results.

[0071] 6. Statistical analysis: Test values ​​and measurements were used in the evaluation even if they appeared to be outliers. If the cause was clearly determined to be a measurement error, they were excluded from the analysis at that point. Missing values ​​were not imputed and were not included in the analysis as missing data.

[0072] The baseline values ​​before the start of the intervention, the measured values ​​at each time point after intake, and the change from baseline were shown as mean ± standard deviation to compare the characteristics of each subject group (gender is the number of subjects). ２A two-sample t-test was used for comparison of changes from baseline between groups and at each post-ingestion time point. A one-sample t-test was used for comparison of changes from baseline in measurements at each post-ingestion time point within groups. For reference, changes at 12 weeks, with baseline set at 0, were evaluated using the Wilcoxon signed-rank test. Similar statistical methods were used for comparisons between and within groups in subgroup analyses. Statistical analysis software used was Microsoft Excel 365 (Microsoft, USA) and IBM SPSS Statistics 26 for Windows (IBM). The statistical significance level was set at 5% on both sides.

[0073] II. Results 1. Subjects Figure 1 shows a study flowchart illustrating the process from subject selection and test food allocation to analysis. A pre-test (blood sampling) was conducted on 264 subjects who provided written consent to participate in the study. A pre-test (cognitive) was conducted on 187 subjects, excluding 72 subjects who were ineligible for study participation and 5 who declined to participate. Based on these results, 100 subjects (50 in each of the test food group and placebo group) who met the inclusion criteria and did not violate any of the exclusion criteria were included in the study as eligible subjects. Subject background information is shown in Table 6. No significant differences were observed between the groups.

[0074]

[0075] 2. Data analysis for primary and secondary endpoints We examined changes after intervention in both groups for each item of the MoCA-J, which was set as the primary endpoint, and for the MMSE and SF-36, which were set as secondary endpoints. Items for which the baseline and change amounts were the same for all participants in both groups were not tested.

[0076] Table 7 shows the total and individual scores for the MoCA-J during the intervention period. Attention (repeated recitation) at week 6 was significantly improved in the placebo group compared with the test food group (P = 0.03). Additionally, total scores, abstract thinking, delayed recall, and orientation were significantly improved at weeks 6 and 12 compared with baseline in both groups.

[0077]

[0078] Regarding the change in the MMSE total score and each item before and after the intervention during the intervention period, significant changes were observed in the total score and question 2 (orientation to place) at 6 and 12 weeks in both groups compared to baseline, with significant increases (improvements); however, no significant differences were observed between the placebo group and the test food group at any of the test weeks.

[0079] Table 8 shows the NBS scores and component summary scores of the SF-36 during the intervention period. At week 12, the test food group showed significant increases (improvements) compared with the placebo group in the NBS scores for mental health, the mental aspect of the 2002 3-component summary score, and the mental aspect of the 1995 2-component summary score (Japanese version) (P = 0.03, 0.01, and 0.03, respectively). Furthermore, the NBS scores for role functioning (mental) and the mental aspect of the 1995 2-component summary score (US version) significantly increased (improved) at week 12 compared with baseline in both groups. The NBS scores for role functioning (physical), the social aspect of the 2002 3-component summary score, and the physical aspect of the 1995 2-component summary score (Japanese version) significantly increased (improved) at week 12 compared with baseline in the placebo group. NBS scores for physical function, vitality and mental health, the mental aspects of the 2002 3-component summary score and the mental aspects of the 1995 2-component summary score (Japanese version) significantly increased (improved) in the test food group at 12 weeks compared to baseline.

[0080]

[0081] 3. Subgroup analysis Regarding the presence or absence of regular exercise habits, all subjects were stratified into those who exercised at least once a week during the intake period (those with regular exercise habits) and those who exercised less than once a week (those without regular exercise habits), and subgroup analysis was performed for each subject stratum. Note that tests were not performed on items where the baseline and change amounts were the same for all subjects in both groups.

[0082] The number of subjects in the placebo group and the test food group in the regular exercise group was 24 (11 men, 13 women) and 26 (15 men, 11 women), respectively, and the number of subjects in the placebo group and the test food group in the non-regular exercise group was 26 (16 men, 10 women) and 24 (11 men, 13 women), respectively.Among the age, male / female ratio, anthropometric measurements, physical examination values, and total MoCA-J score, there was a significant imbalance between the groups in age for the regular exercise group and systolic blood pressure for the non-exercise group (both P = 0.04) (Table 9).

[0083]

[0084] Table 10 shows the total and individual scores for the MoCA-J during the intervention period for those with regular exercise habits. At week 12, the test food group showed a significant increase (improvement) in repetition scores compared with the placebo group (P = 0.03).

[0085]

[0086] In the group with regular exercise habits, no significant differences were observed between the placebo group and the test food group in the change in the MMSE total and individual item scores before and after the intervention period in any of the test weeks.

[0087] In the group without regular exercise habits, no significant differences were observed between the placebo group and the test food group in the changes in the total scores and individual scores of the MoCA-J and MMSE before and after the intervention period in any of the test weeks.

[0088] 4. Safety Evaluation Safety evaluations were conducted on 50 safety analysis subjects in each group (Figure 1) for both adverse events and measurement and test values. During the intervention period, adverse events such as positive urine occult blood tests, diarrhea, joint pain, runny nose, and lower back pain were confirmed in 21 subjects (55 events) in the placebo group and 13 subjects (23 events) in the test food group. Adverse events other than positive urine occult blood tests were either due to an identifiable cause other than the study food or were mild and transient, and were deemed unrelated to the study food, i.e., not to be side effects of the study food, by the investigator. The positive urine occult blood test was considered to be due to the subject's exercise habits, but this could not be identified. Therefore, the investigator determined that it was "probably not" related to the study food, but was within the range of clinically relevant. Significant changes were observed in some measurement and test values ​​when comparing pre- and post-intake values ​​in each group. However, all were within the normal range or minor fluctuations within the reference range, and the investigator determined that they were not clinically relevant.

[0089] III. Discussion This study involved men and women aged 60 years or older who either had subjective symptoms of forgetfulness or had a close relative who had been told they had memory problems. The study involved participants taking a supplement containing DHA and EPA derived from salmon roe oil for 12 weeks, and examining its usefulness on cognitive function and health-related QOL.

[0090] No significant between-group differences were observed in the change in the individual test items or total scores of the MoCA-J and MMSE used to assess cognitive function at 12 weeks. It has been pointed out that in cognitive function assessments, a "practice effect" occurs, whereby memory and familiarity from previous tests result in higher scores on later tests. The total scores for the MoCA-J and MMSE increased compared to baseline in both groups, suggesting that the practice effect influenced the cognitive improvement effects of the test food under the conditions of this study. Furthermore, most subjects achieved the highest scores on some test items, indicating a ceiling effect. Therefore, it is believed that the effects of the test food were not observed in the overall analysis.

[0091] In the "Mental Health" section of the SF-36, which was used to assess health QOL, the change at 12 weeks was 0.11 ± 6.37 points for the placebo group and 2.68 ± 5.20 points for the test food group, demonstrating that the test food significantly improved over placebo. A high score for "Mental Health" indicates that "for the past month, the subject felt calm, happy, and peaceful." The average "Mental Health" score for the subjects in this study was in the 57-point range, which is slightly higher than the average "Mental Health" score of the general Japanese population aged 60 to 84 (NBS score distribution by age and gender in 2017), which is in the 53-54 point range. However, it was found that consuming the test food further improved "Mental Health." Furthermore, regarding the summary score, which is the superordinate concept of the eight subscales, the 2002 3-component summary score "Mental Aspects" and the 1995 2-component summary score (Japanese version) "Mental Aspects" also showed significant improvements at 12 weeks in the test food group compared to the placebo group. The 3-component summary score "Mental Aspects" is composed of "Bodily Pain," "Overall Health," "Social Functioning," "Role Functioning (Mental)," "Vitality," and "Mental Health," while the 2-component summary score "Mental Aspects" is composed of "Overall Health," "Social Functioning," "Role Functioning (Mental)," "Vitality," and "Mental Health." Both of these scores indicate mental QOL. These results suggest that ingesting the test food results in a "calm, happy, and peaceful mood," which is a state of good mental health, and improves mental QOL. Mental health is an important condition for living a vibrant and true life, and specifically includes the ability to recognize and express one's own emotions (emotional health), the ability to think appropriately in the given situation and solve practical problems (intellectual health), the ability to build constructive and good relationships with others and society (social health), and the ability to find purpose and meaning in life and make independent choices about one's life (personal health). It is believed that consuming the test food will improve mental health in these areas as well.

[0092] A subgroup analysis of the test food's effects on cognitive function was conducted based on the presence or absence of regular exercise. Results showed that in subjects who exercised at least once a week (regular exercisers), the test food group showed significant improvements in the MoCA-J "repetition" score compared to the placebo group. Repetition is a test of auditory short-term memory, and the language ability it measures is generally considered to be "the ability to understand the meaning of something, remember it, and accurately communicate it in words." This language ability is said to have three roles: creating something, thinking logically, expressing emotions and feelings, and communicating one's creations, thoughts, emotions, and feelings to others (communication), all of which play an important role in cognitive function. These results suggest that the test food is effective in improving language ability, a part of cognitive function, in elderly people who exercise at least once a week.

[0093] Regarding safety, one case of "positive urine occult blood reaction" was observed in the test food group, which was "probably not" related to the study food, but was judged by the lead investigator to be within the range of clinically relevant. All other adverse events were judged to be "not" related to the study food, and the test food is considered safe to ingest under the conditions of this study.

[0094] Conclusion: When elderly people aged 60 to 85 years old took a supplement containing DHA and EPA derived from salmon roe oil for 12 consecutive weeks, it was suggested that it improved their mental quality of life by making them feel "calm, happy, and peaceful," and that it also improved their language ability, which is one of their cognitive functions, in those who exercise at least once a week. Regarding safety, it was concluded that supplements containing DHA and EPA derived from salmon roe oil can be taken safely under the conditions of this study.

[0095] <Safety evaluation through overdose testing> 1. Subjects The purpose of this study was to confirm the safety of excessive intake (5x dose) of "ONO-SR / AST-SOYPC," a food containing salmon roe oil, astaxanthin, and soy lecithin. When examining food safety, it is desirable to conduct and study across a wide range of age groups and genders. Therefore, in Study 1, to ensure as equal an inclusion of men and women of each age group (20s to 60s), the 5x dose intake group consisted of 20 subjects. For reference, to confirm safety at 1x and 2x doses, the number of subjects was reduced to 5 subjects each in the 1x and 2x dose intake groups. Study 2 was conducted with 10 subjects in the 5x dose intake group only.

[0096] Prior to the study, subjects were given a consent form, the purpose and content of the study were fully explained, and their voluntary consent was obtained in writing. Subjects who gave consent underwent a preliminary examination (lifestyle questionnaire, medical interview, anthropometry, physical examination, and fasting clinical test). Based on the results of the preliminary examination, subjects who did not meet the following exclusion criteria were selected.

[0097] Exclusion criteria: (1) Individuals who regularly consume health foods rich in DHA, EPA, astaxanthin, or soy lecithin; (2) Individuals with or a history of serious diseases such as diabetes, liver disease, kidney disease, or heart disease; (3) Individuals at risk of developing allergies related to the study; (4) Individuals currently undergoing treatment for a disease that may affect the study, or individuals with a history of a chronic or serious disease that required medication; (5) Individuals with a history or current history of drug or alcohol dependence; (6) Individuals who are deemed unsuitable as subjects based on the clinical test values ​​and measurements obtained in the pre-examination; (7) Individuals who have participated in other clinical research within one month of obtaining consent to participate in this study, or individuals who plan to participate in other clinical research after obtaining consent to participate in this study; (8) Individuals who are pregnant, breastfeeding, or plan to become pregnant or breastfeed during the study period; (9) Individuals who are deemed unsuitable as subjects based on the results of the lifestyle questionnaire; (10) Any other person who the responsible physician deems unsuitable as a subject.

[0098] 2. Test Food The test food "ONO-SR / AST-SOYPC" (Ono Pharmaceutical Co., Ltd.) was a soft capsule containing 1g of salmon roe oil (150mg DHA, 60mg EPA), 200μg astaxanthin, 100mg soybean lecithin, and gelatin, glycerin, water, and caramel color as secondary ingredients per 4 capsules. The raw materials and nutritional components are shown in Table 11.

[0099]

[0100] 3. Test Method The test was an open-label study. The 1x dose group took one bag (4 tablets) per day, the 2x dose group two bags (8 tablets), and the 5x dose group five bags (20 tablets) with water or lukewarm water after a meal, without chewing. The time of intake was not specified, and participants were allowed to take the tablets multiple times throughout the day. The intake period was four weeks. During the test period, participants were instructed to make as few changes as possible to their lifestyle habits, such as drinking, eating, and sleeping, to limit excessive exercise that deviated significantly from their normal routine, and to limit dieting and overeating, and not to start any new exercise routines or discontinue any exercise habits they had been maintaining.

[0101] In Study 1, subjects visited the hospital two weeks after taking the test food and underwent a medical interview (to check their physical condition), physical measurements, physical examinations, and fasting clinical tests. Subjects were also instructed to keep a daily diary throughout the study, recording their intake of the test food, changes in their physical condition, mood in daily life, sleep (falling asleep, deep sleep), and medication use. Similar tests were planned for the fourth week of intake and the second week after the end of intake, but due to the declaration of a state of emergency due to COVID-19, subjects were not required to visit the hospital during those two weeks to ensure their safety. After the end of the intake period, subjects continued to keep a diary for two weeks.

[0102] In Study 2, subjects visited the hospital two and four weeks after taking the test food, where they underwent a medical interview (to check their physical condition), physical measurements, physical examinations, and fasting clinical tests. Subjects were also instructed to keep a daily diary throughout the study, recording their intake of the test food, changes in their physical condition, mood in daily life, sleep (how well they fell asleep and slept), medication use, etc. After the end of the intake period, subjects continued to keep the diary for two weeks, and then, two weeks after the end of intake, various tests were conducted as a two-week post-intake test.

[0103] 4. Test items and evaluation items Tests included a medical interview to check physical condition, confirmation of the presence or absence of adverse events, anthropometric measurements (height (pre-test only), weight, BMI), physical examination (systolic and diastolic blood pressure, pulse rate), hematological tests in fasting clinical examinations (white blood cell count, red blood cell count, hemoglobin, hematocrit, platelet count), blood biochemistry tests (total protein, albumin, total bilirubin, direct bilirubin, indirect bilirubin, ALP, AST, ALT, LD, γ-GT, total cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol, urea nitrogen, creatinine, uric acid, sodium, potassium, chloride, blood glucose, HbA1c (pre-test only)), and urine tests (protein qualitative, glucose qualitative, occult blood reaction). In Study 1, all tests were conducted at the pre-test and at week 2 of intake, except for items that were only tested pre-test. In Study 2, with the exception of items that were tested only beforehand, all tests were conducted at the pre-test, 2 weeks after intake, 4 weeks after intake, and 2 weeks after the end of intake. Clinical tests were outsourced to LSI Medience Corporation (Shinagawa-ku, Tokyo).

[0104] Evaluation items were adverse events, measurement values, and test values, and adverse events included subjective symptoms reported in the interview and diary, as well as abnormal fluctuations in test values. Adverse events as subjective symptoms and objective findings were judged by the responsible physician. Abnormal fluctuations in test values ​​(adverse events) for individual subjects were judged as adverse events by the responsible physician based on the reference values ​​set by the medical institution, with reference to the criteria for determining abnormal fluctuations set by the Japanese Society of Chemotherapy, the Common Terminology Criteria for Adverse Events v5.0 Japanese Translation JCOG Edition (CTCAE v5.0-JCOG), and the judgment classification of the Japanese Society of Ningen Dock.

[0105] 5. Statistical analysis The measured values ​​before and after intake at each time point were compared using a one-sample t-test. The significance level of the test was set at 5% on both sides. Microsoft Excel (Microsoft Corporation) was used as the statistical analysis software.

[0106] Results 1) Study 1 1. Subject Background In Study 1, a pre-test was conducted on 63 subjects who had given written consent to participate in the study. Based on the results of the pre-test, 15 men and 15 women, a total of 30 subjects, who met the inclusion criteria but did not meet the exclusion criteria, were selected as eligible subjects for this study. Note that some subjects had clinical test values ​​that exceeded the reference range (test reference values ​​of the clinical testing institution), but in all cases the responsible physician examined each subject individually and, after determining that there were no problems with their participation in the study, they were included in this study.

[0107] All 30 people completed the intake of the test food, but one woman did not visit the hospital due to self-restraint on going out due to the spread of COVID-19, and as there was no data after intake, she was excluded from the analysis due to missing data. Therefore, measurement and test values ​​were evaluated for 29 people (single dose intake group: 5 people, double dose intake group: 5 people, five five-times intake group: 19 people) excluding the one person excluded from the analysis, and adverse events were evaluated for all 30 people as analysis subjects.

[0108] 2. Adverse Events Two cases of fluctuations in measurement values ​​and test results were observed in two subjects. None of the test value fluctuations were deemed abnormal (not adverse events). Adverse events observed included headache, abdominal pain / loose stools, and tendonitis. The incidence and total number of events was one event in one of five subjects (20%) at the 1x dose, two events in two of five subjects (40%) at the 2x dose, and three events in three of 20 subjects (15%) at the 5x dose. All adverse events were mild in severity, and no serious adverse events occurred in this study. Furthermore, it was determined that there was no relationship to the test food.

[0109] 3. Measurements and Test Values ​​There were some significant changes in body measurements, physical examination values, and clinical test values ​​(hematology tests, blood biochemistry tests) after intake compared to before, but the responsible physician determined that all of these changes were at a level that would not pose a clinical problem. Several subjects showed positive clinical test values ​​(urine tests), but this was thought to be due to the influence of menstruation, and the responsible physician determined that there was no clinical problem.

[0110] 2) Study 2 1. Subject background In Study 2, a pre-test was conducted on 23 subjects who had given written consent to participate in the study. Based on the results of the pre-test, 5 men and 5 women, a total of 10 people who met the inclusion criteria and did not meet the exclusion criteria, were selected as eligible subjects for this study.

[0111] All 10 subjects began taking the test food, but one woman was found to be ineligible to be a subject (having a history of cough asthma) on the 15th day of intake and discontinued the study. The remaining 9 subjects completed the prescribed test schedule and content. Therefore, the measurement and test values ​​of the 9 subjects who completed intake, and the adverse events of all 10 subjects were evaluated as analysis subjects.

[0112] 2. Adverse Events Two cases of fluctuation in measurement values ​​and test values ​​were observed in two subjects. Neither of the test value fluctuations was judged to be abnormal (not an adverse event). One case each of diarrhea and sore throat was observed as adverse events, with the frequency and total number of occurrences being two cases in two out of 10 subjects (20%). All adverse events were "mild," and no serious adverse events occurred in this study. Furthermore, it was judged that there was "no" relationship to the test food.

[0113] 3. Measurements and Test Values ​​There were some significant changes in physical measurements, physical examination values, and clinical test values ​​(hematology tests, blood biochemistry tests) after intake compared to before, but the responsible physician determined that all of these changes were at a level that would not pose a clinical problem. Several subjects showed positive clinical test values ​​(urine tests), but this was thought to be due to a transient change, and the responsible physician determined that there was no clinical problem.

[0114] Conclusion The results of the above two studies showed that there were no safety issues when an overdose study was conducted in which participants ingested five times the recommended daily intake of "ONO-SR / AST-SOYPC," a food containing salmon roe oil, astaxanthin, and soy lecithin, for four consecutive weeks.

[0115] The present disclosure relates to improving cognitive function and can be used in the fields of medicine and food.

Claims

1. A composition for improving cognitive function comprising a fish egg lipid preparation that is formulated to provide a daily intake of from about 100 mg to less than 300 mg of DHA, wherein the cognitive function is language ability in a subject who has a regular exercise habit.

2. 2. The composition of claim 1, comprising a fish egg lipid preparation containing at least about 30% phospholipids.

3. The composition according to claim 1, wherein the fish egg lipid preparation contains docosahexaenoic acid (DHA)-bound phospholipids, and the composition ratio of DHA in the constituent fatty acids of the lipid is about 15% or more.

4. The composition of claim 1 further comprising soy lecithin.

5. 2. The composition of claim 1, formulated to provide a daily intake of about 160 mg of DHA.

6. The composition according to claim 1, which is formulated to provide a daily intake of about 100 mg or more but less than 300 mg of DHA, about 30 mg to 300 mg of eicosapentaenoic acid (EPA), and about 50 mg to 500 mg of soy lecithin.

7. 2. The composition of claim 1, formulated to provide a daily intake of about 160 mg of DHA, about 110 mg of EPA, and about 100 mg of soy lecithin.

8. 10. The composition of claim 1 taken daily for at least 12 weeks.

9. The composition according to claim 1, further for improving mental health or mental aspects of QOL.

10. The composition of claim 1 for further improving mental health.

11. 11. The composition of claim 10, wherein improving mental health is improving at least one of nervousness, depression and gloomy mood.

12. The composition according to claim 10, wherein improving mental health is improving to a calm or happy mood.

13. The composition according to claim 9 for improving the mental aspect of QOL as assessed by SF-36 2MCS (Japanese version).

14. The composition according to claim 9 for improving the mental aspect of QOL as determined by the 3MCS of SF-36.

15. The composition according to any one of claims 1 to 14, which is a pharmaceutical composition.

16. The composition according to any one of claims 1 to 14, which is a food composition.