Composition for improving eye disorder

Abstract

The present disclosure provides a composition for improving an eye disorder, said composition comprising a roe lipid preparation which is prepared so as to provide a daily intake of approximately 100-300 mg of docosahexaenoic acid (DHA).

Description

Composition for improving eye conditions

[0001] This patent application claims priority to Japanese Patent Application No. 2023-213159, the entire contents of which are incorporated herein by reference. The present disclosure relates to compositions for improving eye conditions.

[0002] Eye discomfort such as a gritty feeling, pain, difficulty seeing, and blurred vision can be caused by a variety of stimuli, including physical factors (temperature, humidity, ultraviolet rays, etc.), chemical factors (carbon dioxide, cigarette smoke, etc.), and biological factors (pollen, bacteria, viruses), and can limit activities such as reading, driving at night, using a computer, and watching television. Improving eye discomfort in healthy people is thought to play an important role in maintaining and improving health.

[0003] The Ministry of Health, Labour and Welfare published the VDT Guidelines in 2002, which set out precautions for workers who use computers and other devices. In particular, in recent years, with the advent of IT devices such as personal computers, smartphones, and tablets, staring at screens for long periods of time has caused eye strain, raising concerns that this can have a negative impact on not only physical health but also mental health. The 2019 guidelines were significantly updated, highlighting the importance of managing eye health as well as managing the work environment when using IT devices.

[0004] The physiological activities of ω-3 polyunsaturated fatty acids such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) have recently attracted attention, and it has been reported that they improve cognitive function and sleep quality (Patent Documents 1 and 2). Patent Document 3 discloses a composition rich in DHA-bound phospholipids.

[0005] Regarding dry eye, it has been reported that 30 days of ingestion of 240 mg DHA / 360 mg EPA significantly improved the Ocular Surface Disease Index (OSDI) (Non-Patent Document 1), 3 months of ingestion of 350 mg DHA / 650 mg EPA significantly improved the subjective symptom score of the eye (Non-Patent Document 2), 90 days of ingestion of krill oil containing 510 mg DHA / 945 mg EPA or fish oil containing 500 mg DHA / 1000 mg EPA significantly improved the OSDI with krill oil (Non-Patent Document 3), and 12 weeks of ingestion of fish oil containing 540 mg DHA / 1245 mg EPA improved eye pain (Non-Patent Document 4). In these reports, in addition to improvement of subjective symptoms, improvement effects were also observed in objective findings such as tear film break-up time (BUT) and Schirmer test. On the other hand, Non-Patent Document 5 reports that when 506 mg of re-esterified triglyceride-type DHA / 1680 mg of EPA was administered for two months, subjective symptoms measured by OSDI and Dry Eye-Related Quality-of-Life Score (DEQS) were significantly improved, but BUT and Schirmer test scores were not improved.

[0006] International Publication No. 2022 / 210856 Pamphlet International Publication No. 2022 / 244727 Pamphlet International Publication No. 2021 / 132516 Pamphlet

[0007] Kangari H, Eftekhari MH, Sardari S, Hashemi H, Salamzadeh J, Ghassemi-Broumand M, et al:Short-term consumption of oral omega-3 and dry eye syndrome. Ophthalmology 2013; 120:2191-2196Bhargava R, Kumar P, Kumar M, Mehra N, Mishra A. A randomized controlled trial of omega-3 fatty acids in dry eye syndrome. Int J Ophthalmol, 2013;6(6):811-816Deinema LA, Vingrys AJ, Wong CY, et al. A randomized, double-masked, placebo-controlled clinical trial of two forms of omega-3 supplements for treating dry eye disease. Ophthalmology 2017;124:43-52Kawakita T, Kawabata F, Tsuji T, Kawashima M, Shimmura S, Tsubota K. Effects of dietary supplementation with fish oil on dry eye syndrome subjects: randomized controlled trial. Biomed Res. 2013; 34: 215-20Park J, Yoo YS, Shin E, Han G, Shin K, Lim DH, Chung TY. Effects of the re-esterified triglyceride (rTG) form of omega-3 supplements on dry eye following cataract surgery. Br J Ophthalmol. 2021; 105: 1504-1509

[0008] An object of the present disclosure is to provide a new means for improving eye conditions.

[0009] The present disclosure provides a composition for improving eye complaints, comprising a fish roe lipid preparation.

[0010] The present disclosure provides a new means for improving eye conditions.

[0011] This figure shows a flowchart of subject follow-up in the study of the example. The figures show the change in J-OSDI and DEQS scores before and after 12 weeks of placebo or test food intake. A) J-OSDI total score. The sub-item eye symptom score and environmental factor score significantly decreased and improved in the test food intake group (●) compared to the placebo intake group (◯). B) DEQS summary score. The sub-item eye symptom score tended to decrease in the test food intake group compared to the placebo intake group. Data analysis was performed using the Manne-Whitney U test. * indicates the result of a significant difference between the two groups. *p<0.05. The figures show the change in DHA-PC (38:6-PC, 40:6-PC, 40:7-PC, and 44:12-PC) in human plasma before and after 12 weeks of placebo or test food intake. Plasma DHA-PC levels were significantly elevated in the test food intake group compared to the placebo intake group. Data analysis was performed using a multiple t-test. *** in the figure indicates the results of a significant difference test between the two groups. ***p<0.005

[0012] 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.

[0013] 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.

[0014] (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) or kunimasu (Oncorhynchus kawamurae). Examples of Atlantic salmon include Atlantic salmon (Salmo salar) or brown trout (Salmo trutta). In one embodiment, the fish roe lipid preparation is prepared from pink salmon (Oncorhynchus gorbuscha) roe.

[0015] 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.

[0016] 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 with 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). Alternatively, 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.

[0017] As the fish roe lipid preparation, the fish roe lipid composition described in WO2021 / 132516 (Patent Document 3) may be used.

[0018] (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 are 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 are sphingomyelin (SM) and / or 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.

[0019] 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%.

[0020] 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%.

[0021] 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.

[0022] (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.

[0023] 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 12% or more.

[0024] 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) or docosapentaenoic acid (DPA) (C22:5). Other constituent fatty acids may 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), or docosadienoic acid (C22:2).

[0025] The weight of DHA per 100 g of fish roe lipid preparation is, for example, about 10 g or more, preferably about 12 g or more, more preferably about 14 g or more, and even more preferably about 15 g or more. The upper limit of the weight of DHA per 100 g of fish roe lipid preparation is not particularly limited, but is, for example, about 30 g or less, preferably about 25 g or less, more preferably about 22 g or less, and even more preferably about 20 g or less.

[0026] 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.

[0027] Fish egg lipid preparations contain many phospholipids bound with DHA. Fish egg lipid preparations may contain phosphatidylcholine having at least one DHA bound as a constituent fatty acid (DHA-bound PC). Examples of DHA-bound PC include palmitoyl-docosahexanoyl-glycerophosphocholine, stearoyl-docosahexanoyl-glycerophosphocholine, oleoyl-docosahexanoyl-glycerophosphocholine, and / or didocosahexanoyl-glycerophosphocholine.

[0028] One of the DHA-bound PCs contained in fish egg lipid preparations is palmitoyl-docosahexanoyl-glycerophosphocholine (PDPC). PDPC refers to phosphatidylcholine (38:6) in which one of the C-1 and C-2 acyl groups is hexadecanoyl (16:0) and the other is docosahexanoyl (22:6). Herein, palmitoyl-docosahexanoyl-glycerophosphocholine (PDPC) is sometimes referred to as 38:6-PC.

[0029] One of the DHA-bound PCs contained in fish egg lipid preparations is stearoyl-docosahexanoyl-glycerophosphocholine (SDPC). SDPC refers to phosphatidylcholine (40:6) in which one of the C-1 and C-2 acyl groups is stearoyl (18:0) and the other is docosahexanoyl (22:6). In this specification, stearoyl-docosahexanoyl-glycerophosphocholine (SDPC) is sometimes referred to as 40:6-PC.

[0030] One of the DHA-bound PCs contained in fish egg lipid preparations is oleoyl-docosahexanoyl-glycerophosphocholine. Oleoyl-docosahexanoyl-glycerophosphocholine refers to phosphatidylcholine (40:7) in which one of the C-1 and C-2 acyl groups is oleoyl (18:1) and the other is docosahexanoyl (22:6). In this specification, oleoyl-docosahexanoyl-glycerophosphocholine is sometimes referred to as 40:7-PC.

[0031] One of the DHA-bound PCs contained in fish egg lipid preparations is didocosahexanoyl-glycerophosphocholine. Didocosahexanoyl-glycerophosphocholine refers to phosphatidylcholine (44:12) in which both the C-1 and C-2 acyl groups are docosahexanoyl (22:6). In this specification, didocosahexanoyl-glycerophosphocholine is sometimes referred to as 44:12-PC.

[0032] 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.

[0033] In one embodiment, the fish roe lipid preparation comprises any one selected from the group consisting of DHA-linked ether-type phospholipids and EPA-linked ether-type phospholipids.

[0034] (Components other than lipids) The fish roe lipid preparation may contain components other than lipids. Components other than lipids include proteins and inorganic substances such as sodium, potassium, or phosphorus. Lipids refer to biologically derived substances that are soluble in non-polar solvents, and include simple lipids, complex lipids, or 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.

[0035] 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.

[0036] (Uses) As shown in the examples below, ingestion of a composition of the present disclosure can improve eye complaints. Eye complaints are subjective symptoms caused by eye fatigue or dryness, and include, for example, eye discomfort such as dry eye (dryness), foreign body sensation (gritty sensation), blurred vision (hazy vision), difficulty in seeing, photophobia (glare), eye fatigue, heavy eye sensation, eye pain (stinging eye sensation), or eye itching. Eye complaints may be chronic or temporary. In some embodiments, the eye complaint is discomfort caused by eye sensitivity due to temporary dryness (e.g., in windy conditions, low humidity, or air-conditioned areas). In some embodiments, the eye complaint is dry eye or eye fatigue. In some embodiments, the eye complaint is photophobia, foreign body sensation, eye pain, blurred vision, or difficulty in seeing.

[0037] Eye complaints can be assessed, for example, by the Ocular Surface Disease Index (OSDI), the Japanese version of the Ocular Surface Disease Index (J-OSDI), or the Dry Eye-Related Quality-of-Life Score (DEQS).

[0038] It is known that objective findings and subjective symptoms can be dissociated in dry eye patients. For example, patients may experience severe pain even when there are no abnormalities in the corneal and conjunctival epithelium or tear film. Ingestion of a composition of the present disclosure may or may not improve objective findings related to the eyes. Objective findings can be evaluated, for example, by the tear film break-up time (BUT) test or the Schirmer test.

[0039] In the examples described below, ingestion of a composition of the present disclosure significantly improved the J-OSDI total score, sub-item ocular symptom score, and environmental factor score, which are indicators of subjective dry eye symptoms, but no difference was observed in BUT, an indicator of objective findings. Therefore, without being limited by theory, it is believed that the composition of the present disclosure has a sensory nerve improving effect and improves eye disorders through the improvement of sensory nerves. Improvement of sensory nerves includes improving, reducing, or alleviating sensory nerve hypersensitivity, or restoring sensory nerve hypersensitivity to normal. For example, the eye disorder may be an eye disorder in a subject with sensory nerve hypersensitivity, an eye disorder in a subject with temporarily increased sensory nerve sensitivity, or an eye disorder in a situation where the sensory nerves of the eye become sensitive due to temporary dryness.

[0040] Eye disorders can cause physical stress. Examples of physical stress include difficulty recovering from fatigue, fatigue, mental clarity, eye fatigue, strain on the shoulders and neck, and discomfort in the lower back and back. Therefore, in some embodiments, the compositions of the present disclosure can reduce physical stress caused by eye disorders. Physical stress can be assessed, for example, using a stress assessment questionnaire (PHRFSCL-SF).

[0041] The subject is typically a human. The subject's age is not limited, but may typically be 20 years or older, 25 years or older, 30 years or older, 35 years or older, or 40 years or older, and may be under 85 years, under 80 years, under 75 years, or under 70 years old. For example, the subject's age is 40 years or older but under 70 years old. The subject may have a temporary eye disorder, such as temporary eye fatigue or dryness due to their living environment. The subject may not have been diagnosed with an eye disease, but may be a patient with an eye disease, including dry eye. In some embodiments, the subject is a patient with subjective symptoms of dry eye. In some embodiments, the subject is a patient with subjective symptoms of dry eye but no objective findings. For example, the presence of depression or anxiety symptoms may result in a high dry eye symptom score, while the objective findings may not differ from those of healthy individuals. Thus, in some embodiments, the subject is a dry eye patient with depression or anxiety symptoms.

[0042] 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, or emulsions. Dosage forms for transdermal administration include patches, tapes, sprays, lotions, creams, ointments, liquids, emulsions, or suspensions. Dosage forms for nasal administration include nasal drops or nasal sprays.

[0043] 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.

[0044] 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, it may be a food to be consumed as a solid food or a liquid such as a beverage, a drinkable supplement, a powdered beverage, or a soup. Specifically, it may be consumed as, for example, juice, confectionery, jelly, tablet, dressing, or seasoning.

[0045] 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 eye disorders, improving the severity of eye disorders, or reducing physical stress caused by eye disorders. The labeling may be directly displayed on the packaging, container, label, tag, or attached documentation accompanying the product, or indirectly displayed through advertising and promotional activities. Dietary supplements include, for example, nutritional supplements and health supplements.

[0046] 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.

[0047] 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 about 300 mg or less, and especially preferably about 200 mg or less. For example, the amount of intake of the composition of the present disclosure can be an amount that results in about 100 mg to 300 mg of DHA being ingested per day, preferably about 150 mg to 200 mg of DHA being ingested per day. 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) of DHA being ingested per day.

[0048] In some embodiments, the composition of the present disclosure can be ingested in an amount that results in about 10 mg or more of DHA-bound PC per day, preferably about 30 mg or more, more preferably about 50 mg or more, and even more preferably about 70 mg or more. The composition of the present disclosure can be ingested in an amount that results in about 1,000 mg or less of DHA-bound PC per day, preferably about 500 mg or less, more preferably about 300 mg or less, even more preferably about 200 mg or less, even more preferably about 150 mg or less, and especially preferably about 100 mg or less. For example, the composition of the present disclosure can be ingested in an amount that results in about 50 mg to 150 mg of DHA-bound PC per day, preferably about 70 mg to 100 mg. In some embodiments, the composition of the present disclosure can be ingested in an amount that results in about 80 mg (e.g., 72 mg to 88 mg) of DHA-bound PC per day.

[0049] The intake amount 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 intake amount 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. For example, the intake amount of the composition of the present disclosure can be an amount that results in an intake of about 30 mg to 300 mg of EPA per day, preferably about 40 mg to 200 mg, and more preferably about 50 mg to 150 mg. In some embodiments, the intake amount 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) or about 110 mg (e.g., 99 mg to 121 mg) of EPA per day.

[0050] 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.

[0051] The compositions of the present disclosure may be taken in a single dose or multiple doses. When taken multiple times, the compositions may be taken, for example, once to several times a day, e.g., once, twice, three or four times a day, daily or every few days, e.g., every 1, 2, 3 or 7 days. The compositions of the present disclosure may be taken at any time, regardless of meal or sleep times. The duration of intake is not limited, and the compositions may be taken continuously for a period of time required to improve the eye condition, for example, for one week or more, preferably one month or more, more preferably two months or more, and particularly preferably three months or more. There may be periods during which the intake is discontinued. In some embodiments, the compositions of the present disclosure are taken in divided doses, one to four times a day. In some embodiments, the compositions of the present disclosure are taken once a day. In some embodiments, the compositions of the present disclosure are taken daily for at least 12 weeks.

[0052] 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.

[0053] The composition of the present disclosure may further contain astaxanthin. The daily intake of astaxanthin may be about 50 μg or more, preferably about 100 μg or more, more preferably about 200 μg or more, and may be 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 of two or more.

[0054] 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 krill-derived cellulose include Astax-S manufactured by Marine Daio Co., Ltd.

[0055] In one aspect, a composition for improving eye disorders is provided, the composition comprising a fish roe lipid preparation. In one aspect, a method for improving eye disorders is provided, the method comprising administering a fish roe lipid preparation to a subject in need thereof. In one aspect, a fish roe lipid preparation for improving eye disorders is provided. In one aspect, a use of a fish roe lipid preparation for improving eye disorders is provided. In one aspect, a use of a fish roe lipid preparation in the manufacture of a composition for improving eye disorders is provided.

[0056] In one aspect, a composition for improving ocular sensory nerves is provided, comprising a fish roe lipid preparation. Other ingredients that may be included in the composition and methods for using the composition are the same as those described above. In one aspect, a method for improving ocular sensory nerves is provided, comprising ingesting a fish roe lipid preparation to a subject in need of such improvement. In one aspect, a fish roe lipid preparation for improving ocular sensory nerves is provided. In one aspect, a use of ... in the manufacture of a composition for improving ocular sensory nerves is provided.

[0057] For example, the following embodiments are provided: [1] A composition for improving eye disorders, comprising a fish roe lipid preparation. [2] The composition according to item 1, which is formulated to provide a daily intake of about 100 mg to 300 mg (preferably about 150 mg to 200 mg) of docosahexaenoic acid (DHA). [3] The composition according to item 1 or 2, which is formulated to provide a daily intake of about 150 mg (or 135 mg to 165 mg) of DHA. [4] The composition according to any one of items 1 to 3, wherein the fish roe lipid preparation comprises phosphatidylcholine having at least one DHA bound thereto (DHA-bound PC) as a constituent fatty acid. [5] The composition according to item 4, wherein the DHA-bound PC comprises palmitoyl-docosahexanoyl-glycerophosphocholine, stearoyl-docosahexanoyl-glycerophosphocholine, oleoyl-docosahexanoyl-glycerophosphocholine, didocosahexanoyl-glycerophosphocholine, or a combination thereof. [6] The composition according to any one of items 1 to 5, which is formulated to provide a daily intake of about 50 mg to 150 mg (preferably about 70 mg to 100 mg) of DHA-bound PC. [7] The composition according to any one of items 1 to 6, which is formulated to provide a daily intake of about 80 mg (or 72 mg to 88 mg) of DHA-bound PC. [8] The composition according to any one of items 1 to 7, wherein the fish roe lipid preparation contains eicosapentaenoic acid (EPA). [9] The composition according to any one of items 1 to 8, which is formulated to provide a daily intake of about 30 mg to 300 mg (preferably about 40 mg to 200 mg, more preferably about 50 mg to 150 mg) of EPA.

[10] The composition according to any one of items 1 to 9, which is formulated to provide a daily intake of about 60 mg (or 54 mg to 66 mg) of EPA.

[0058]

[11] The composition according to any one of items 1 to 9, which is formulated to provide a daily intake of about 110 mg (or 99 mg to 121 mg) of EPA.

[12] The composition according to any one of items 1 to 11, which further contains astaxanthin.

[13] The composition according to item 12, wherein the astaxanthin is derived from a pigment of Haematococcus algae.

[14] The composition according to any one of items 1 to 13, which is formulated to provide a daily intake of about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg) of astaxanthin.

[15] The composition according to any one of items 1 to 14, which is formulated to provide a daily intake of about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[16] The composition according to any one of items 1 to 15, which is prepared so that the daily intake amounts are about 100 mg to 300 mg (preferably about 150 mg to 200 mg) of DHA, about 30 mg to 300 mg (preferably about 40 mg to 200 mg, more preferably about 50 mg to 150 mg) of EPA, and about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg) of astaxanthin.

[17] The composition according to any one of items 1 to 16, which is prepared so that the daily intake is about 100 mg to 300 mg (preferably about 150 mg to 200 mg) of DHA, about 50 mg to 150 mg (preferably about 70 mg to 100 mg) of DHA-bound PC, about 30 mg to 300 mg (preferably about 40 mg to 200 mg, more preferably about 50 mg to 150 mg) of EPA, and about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg) of astaxanthin.

[18] The composition according to any one of items 1 to 17, which is prepared so that the daily intake is about 150 mg (or 135 mg to 165 mg) of DHA, about 60 mg (or 54 mg to 66 mg) or about 110 mg (or 99 mg to 121 mg) of EPA, and about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[19] The composition according to any one of items 1 to 18, which is formulated to provide a daily intake of about 150 mg (or 135 mg to 165 mg) of DHA, about 80 mg (or 72 mg to 88 mg) of DHA-bound PC, about 60 mg (or 54 mg to 66 mg) or about 110 mg (or 99 mg to 121 mg) of EPA, and about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[20] The composition according to any one of items 1 to 18, 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 200 μg (or 180 μg to 220 μg) of astaxanthin.

[0059]

[21] The composition according to any one of items 1 to 20, which is formulated to provide a daily intake of about 150 mg (or 135 mg to 165 mg) of DHA, about 80 mg (or 72 mg to 88 mg) of DHA-bound PC, about 60 mg (or 54 mg to 66 mg) of EPA, and about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[22] The composition according to any one of items 1 to 21, wherein the eye complaint is dry eye, foreign body sensation, blurred vision, difficulty in seeing, photophobia, eye fatigue, heavy eye sensation, eye pain, or itchy eye.

[23] The composition according to any one of items 1 to 22, wherein the eye complaint is photophobia, foreign body sensation, eye pain, blurred vision, or difficulty in seeing.

[24] The composition according to any one of items 1 to 23, wherein the eye complaint is dry eye or eye fatigue.

[25] The composition according to any one of items 1 to 24, which further reduces physical stress caused by eye disorders.

[26] The composition according to any one of items 1 to 25, which improves eye disorders through improvement of sensory nerves.

[27] A composition for improving eye sensory nerves, comprising a fish roe lipid preparation.

[28] The composition according to item 27, which is formulated to provide a daily intake of about 100 mg to 300 mg (preferably about 150 mg to 200 mg) of docosahexaenoic acid (DHA).

[29] The composition according to item 27 or 28, which is formulated to provide a daily intake of about 150 mg (or 135 mg to 165 mg) of DHA.

[30] The composition according to any one of items 27 to 29, wherein the fish roe lipid preparation comprises phosphatidylcholine having at least one DHA bound thereto (DHA-bound PC) as a constituent fatty acid.

[0060]

[31] The composition according to item 30, wherein the DHA-bound PC comprises palmitoyl-docosahexanoyl-glycerophosphocholine, stearoyl-docosahexanoyl-glycerophosphocholine, oleoyl-docosahexanoyl-glycerophosphocholine, didocosahexanoyl-glycerophosphocholine, or a combination thereof.

[32] The composition according to any one of items 27 to 31, which is formulated to provide a daily intake of about 50 mg to 150 mg (preferably about 70 mg to 100 mg) of DHA-bound PC.

[33] The composition according to any one of items 27 to 32, which is formulated to provide a daily intake of about 80 mg (or 72 mg to 88 mg) of DHA-bound PC.

[34] The composition according to any one of items 27 to 33, wherein the fish roe lipid preparation contains eicosapentaenoic acid (EPA).

[35] The composition according to any one of items 27 to 34, which is formulated to provide a daily intake of about 30 mg to 300 mg (preferably about 40 mg to 200 mg, more preferably about 50 mg to 150 mg) of EPA.

[36] The composition according to any one of items 27 to 35, which is formulated to provide a daily intake of about 60 mg (or 54 mg to 66 mg) of EPA.

[37] The composition according to any one of items 27 to 35, which is formulated to provide a daily intake of about 110 mg (or 99 mg to 121 mg) of EPA.

[38] The composition according to any one of items 27 to 37, which further comprises astaxanthin.

[39] The composition according to item 38, wherein the astaxanthin is derived from a pigment of Haematococcus algae.

[40] The composition according to any one of items 27 to 39, which is prepared so that the daily intake of astaxanthin is about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg).

[0061]

[41] The composition according to any one of items 27 to 40, which is formulated to provide a daily intake of about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[42] The composition according to any one of items 27 to 41, which is formulated to provide a daily intake of about 100 mg to 300 mg (preferably about 150 mg to 200 mg) of DHA, about 30 mg to 300 mg (preferably about 40 mg to 200 mg, more preferably about 50 mg to 150 mg) of EPA, and about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg) of astaxanthin.

[43] The composition according to any one of items 27 to 42, which is prepared so that the daily intake is about 100 mg to 300 mg (preferably about 150 mg to 200 mg) of DHA, about 50 mg to 150 mg (preferably about 70 mg to 100 mg) of DHA-bound PC, about 30 mg to 300 mg (preferably about 40 mg to 200 mg, more preferably about 50 mg to 150 mg) of EPA, and about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg) of astaxanthin.

[44] The composition according to any one of items 27 to 43, which is prepared so that the daily intake is about 150 mg (or 135 mg to 165 mg) of DHA, about 60 mg (or 54 mg to 66 mg) or about 110 mg (or 99 mg to 121 mg) of EPA, and about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[45] The composition according to any one of items 27 to 44, which is prepared so that the daily intake is about 150 mg (or 135 mg to 165 mg) of DHA, about 80 mg (or 72 mg to 88 mg) of DHA-bound PC, about 60 mg (or 54 mg to 66 mg) or about 110 mg (or 99 mg to 121 mg) of EPA, and about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[46] The composition according to any one of items 27 to 44, 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 200 μg (or 180 μg to 220 μg) of astaxanthin.

[47] The composition according to any one of items 27 to 46, which is formulated to provide a daily intake of about 150 mg (or 135 mg to 165 mg) of DHA, about 80 mg (or 72 mg to 88 mg) of DHA-bound PC, about 60 mg (or 54 mg to 66 mg) of EPA, and about 200 μg (or 180 μg to 220 μg) of astaxanthin.

[48] The composition according to any one of items 1 to 47, which is intended to be taken by a subject aged 40 to under 70 years.

[49] The composition according to any one of items 1 to 48, which is taken in one to four divided doses per day.

[50] The composition according to any one of items 1 to 49, which is taken once per day.

[0062]

[51] The composition according to any one of items 1 to 50, which is taken daily for at least 12 weeks.

[52] The composition according to any one of items 1 to 51, which is a pharmaceutical composition.

[53] The composition according to any one of items 1 to 51, which is a food composition.

[54] A method for improving eye disorders, comprising ingesting a fish roe lipid preparation to a subject in need thereof.

[55] A fish roe lipid preparation for improving eye disorders.

[56] Use of a fish roe lipid preparation for improving eye disorders.

[57] Use of a fish roe lipid preparation in the manufacture of a composition for improving eye disorders.

[58] A method for improving ocular sensory nerves, comprising ingesting a fish roe lipid preparation to a subject in need thereof.

[59] A fish roe lipid preparation for improving ocular sensory nerves.

[60] Use of a fish roe lipid preparation for improving ocular sensory nerves.

[61] Use of a fish roe lipid preparation in the manufacture of a composition for improving ocular sensory nerves.

[0063] All documents cited in this specification are incorporated herein by reference. 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. The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0064] <Production of fish roe lipid preparation from salmon roe> 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 resulting filtrate under reduced pressure, producing a composition (40 PL) (Table 1) containing approximately 40% phospholipids as a red oily substance.

[0065] *Quantitative values ​​are the total amount of lipids containing each fatty acid and free fatty acids.

[0066] <Production of Soft Capsules> Haematococcus algae pigment (astaxanthin) was added to the 40PL to produce soft capsules "ONO-SR / AST" containing the ingredients shown in Table 2. The daily intake of salmon roe oil was 1,000 mg. Placebo capsules containing safflower oil instead of 40PL and astaxanthin were also produced.

[0067] The amounts of phospholipid classes contained in ONO-SR / AST are shown in Table 3.

[0068] The specifications of ONO-SR / AST are as shown in the table below.

[0069] <Clinical trial> Subjects and methods 1. Subjects This study targeted healthy Japanese men and women aged 40 to under 70, and Imec RD Co., Ltd. publicly recruited paid volunteers. Recruitment was carried out after approval by the Institutional Review Board. The responsible physician provided all potential subjects with informed consent forms and fully explained the purpose and content of the study. After the explanation was completed, written consent to participate in the study based on each individual's free will was obtained. Eligible subjects were selected from the potential subjects for whom this consent was obtained.

[0070] The inclusion criteria were as follows: (1) Age: 40 years or older and younger than 70 years old (2) Gender: Japanese men and women (3) BMI less than 30.0 (4) Those experiencing dry eyes or eye fatigue (5) Those who do not wear contact lenses or those who have switched to glasses two weeks prior to the screening test and will not wear contact lenses during the study period (6) Those who are able to enter an electronic diary using a smartphone or PC (7) Those who have received a full explanation of the purpose and content of the study, fully understood it, and voluntarily volunteered to participate, and provided written consent to participate in the study.

[0071] The exclusion criteria were as follows: (1) Subjects currently receiving outpatient or drug or herbal treatment for any illness (occasional use is acceptable) (2) Subjects currently undergoing eye-related treatment (3) Subjects undergoing dietary therapy and exercise therapy under the supervision of a doctor (4) Subjects who have undergone eye surgery within one year prior to the screening test date or who are scheduled to undergo eye surgery during the study period (5) Subjects who have used eye drops, including over-the-counter (OTC) medications, within two weeks prior to the screening test date (6) Subjects with serious or progressive illnesses or symptoms, or those with a history of serious illnesses, and those who cannot avoid using eye drops, including OTC medications, during the study period (7) Subjects with hay fever or allergic conjunctivitis (8) Subjects who regularly consume yogurt or lactic acid bacteria drinks (subjects who can stop after obtaining consent are eligible to participate) (9) Subjects who regularly consume health foods rich in DHA, EPA, and astaxanthin at least once a week (10) Subjects who consume specific foods for eye health at least once a week (blueberries, purple sweet potatoes, etc.) (11) Individuals who have the habit of consuming fish and fish eggs four or more times a week. (12) Individuals who currently regularly use over-the-counter medicines and quasi-drugs, foods for specified health uses, health foods, or supplements. (13) Individuals with current or past drug or food allergies. (14) Individuals who plan to make major changes to their lifestyle (diet, sleep, exercise, etc.) during the study period. (15) Individuals who are pregnant, breastfeeding, or wish to become pregnant during the study period. (16) Individuals who work night shifts or other shift work. (17) Individuals who have participated in, are currently participating in, or plan to participate in other clinical trials within one month prior to obtaining consent during the study period. (18) Individuals who the investigator deems unsuitable for participation in this study.

[0072] The subjects were required to adhere to the following restrictions. [1] During the study period: (1) Intake of health foods and supplements is prohibited. (2) Use of eye drops is prohibited. (3) Use of contact lenses is prohibited. (4) Massage or cold / hot therapy (including hot eye masks) is not permitted to treat eye fatigue. (5) Visit the hospital at the designated time. (6) Take the test food as instructed. (7) Do not let anyone else take the test food. (8) Enter data into an electronic diary every day. (9) Do not consume yogurt, lactic acid bacteria drinks, blueberries, purple sweet potatoes, etc. (10) Do not consume fish or fish eggs more than four times a week. (11) If you experience any physical abnormalities during the study period, immediately report them to the principal investigator and follow their instructions. (12) Except in emergencies, use only medications approved by the principal investigator. If medications are used, enter the reason for use, the name of the medication, the amount used, the period of use, etc. into the electronic diary and contact the study personnel. (13) If you are unable to attend the hospital for any reason, contact the study staff immediately. (14) Do not participate in any studies involving the ingestion of other foods or drugs, or studies involving the application of cosmetics or drugs. (15) Live the same lifestyle as before participating in the study. In particular, do not engage in binge eating, excessive dietary restrictions, change your diet when traveling abroad, suddenly stop exercising, start a new exercise program, stay up too late, or change your drinking habits. (16) If the test results in a positive result for COVID-19, contact the study's principal physician immediately. (17) Do not disclose any information about the study or test foods to others. [2] The day before visiting the hospital: (1) Abstain from alcohol from the day before each test until the end of the test. (2) Finish eating by 10:00 p.m. [3] The day of visiting the hospital: (1) Do not eat or drink anything other than water until the end of the test. (2) Do not engage in any visual activity, such as reading, working on a computer, or playing video or mobile games, until the end of the test.

[0073] 2. Test Food The test food was formulated with the daily intake amount (one bag, four tablets) of salmon roe oil (functional ingredients: 80 mg DHA-bound PC, 150 mg DHA, and 60 mg EPA) and 4 mg Haematococcus algae pigment (equivalent to 200 μg astaxanthin). Astaxanthin was added as a stabilizer for unsaturated fatty acids such as DHA. The placebo contained safflower oil instead of salmon roe oil, and caramel coloring was added to make it indistinguishable from the test food in appearance. The test food and placebo were provided by Ono Pharmaceutical Co., Ltd. Four tablets of the test food and placebo were taken with water once a day after breakfast. If subjects forgot to take the test food or placebo after breakfast, they were to take it on the same day and were prohibited from carrying it over to the next day.

[0074] 3. Study design This study was designed as a randomized, double-blind, placebo-controlled, parallel-group comparative study. Volunteers were given informed consent during the pre-test, and the pre-test was conducted on 86 subjects who provided written consent to participate in the study. Based on the results of the pre-test, subjects who met the inclusion criteria but did not meet the exclusion criteria were selected, and eligibility was determined based on the overall judgment of the principal investigator, with 44 subjects being included in this study.

[0075] Regarding the number of patients, an effect size (Hedge's g) of 0.898 was calculated based on data from a previous study (Kawakita T, Kawabata F, Tsuji T, Kawashima M, Shimmura S, Tsubota K. Effects of dietary supplementation with fish oil on dry eye syndrome subjects: randomized controlled trial. Biomed Res. 2013; 34: 215-20) on subjective symptom VAS assessment of eye pain at 12 weeks. Based on the calculated effect size, the required sample size for subjective assessment of eye function was estimated to be 20 patients per group, assuming a significance level of α = 0.05 and a statistical power of 80% using a two-tailed, two-sample t-test. Furthermore, assuming a 10% discontinuation or dropout rate during the study period, a total of 44 patients (22 patients per group) was determined.

[0076] The allocation factors were age, sex, BMI, and BUT test results, and subjects were divided into two groups by stratified randomization with a 1:1 ratio of placebo intake group to test food intake group. Allocation was carried out by an allocation officer at Imec RD Co., Ltd., who was not involved in this study. Allocation information was strictly stored until the key was opened after the cases were fixed, maintaining blinding for all parties involved and subjects except the allocation officer.

[0077] The test food was continuously taken from May 14, 2022 to August 7, 2022. Various tests were performed from week 0 to week 12 after intake (Table 5). All tests were performed at National Sakura Hospital, a medical corporation. General blood tests were performed by BML Co., Ltd.

[0078]

[0079] 5. Evaluation Methods A. Primary Evaluation Items 1) J-OSDI (Japanese version of the Ocular Surface Disease Index) The J-OSDI (Midorikawa-Inomata A, Inomata T, Nojiri S, Nakamura M, Iwagami M, Fujimoto K, Okumura, et al. Reliability and validity of the Japanese version of the Ocular Surface Disease Index for dry eye disease. BMJ Open 2019; 9: e033940.) is the Japanese version of the OSDI, the most widely used questionnaire in many dry eye studies. It consists of 12 items with three sub-scores: ocular symptoms, vision-related function (impact on daily life), and environmental factors. It is also considered useful for diagnosing and classifying the severity of dry eye, and is simple and easy to use.

[0080] The J-OSDI (Japan Dry Eye Survey Inventory) was administered before and at the 12-week visit. To what extent have you experienced the following symptoms in the past week? 1. Glare, 2. Gritty eyes, 3. Eye pain, 4. Blurred vision, 5. Difficulty seeing. To what extent have your eye symptoms limited the following activities in the past week? 6. Reading, 7. Driving at night, 8. Using a computer, bank, or ATM, 9. Watching television. In the past week, have you experienced eye discomfort in the following environments? 10. When the wind is strong, 11. In places with low humidity (dry), 12. In places with air conditioning. Participants were asked to answer "Always (4), Most of the time (3), Half the time (2), Sometimes (1), Never (0), or Not applicable (-)." Responses to questions 1 through 5 were summed to calculate an eye symptom score. Responses to questions 6 through 9 were summed to calculate a vision-related function score. The responses to questions 10 to 12 were summed to calculate the environmental factor score. The responses to all questions were summed to calculate the total score. The 12-week change in each sub-item and total score was calculated by subtracting the pre-intake measurement value from the 12-week post-intake measurement value.

[0081] 2) DEQS (Dry Eye-Related Quality-of-Life Score) The development process of the DEQS (Sakane Y, Yamaguchi M, Yokoi N, Uchino M, Dogru M, Oishi T, et al. Development and Validation of the Dry Eye-Related Quality-of-Life Score Questionnaire. JAMA Ophthalmol. 2013; 131:1331-1338) has been published in a paper, and its reliability and validity have been verified psychometrically. It consists of six items related to ocular symptoms and nine items related to interference with daily life, and is said to be useful for evaluating changes in symptoms and QOL due to treatment.

[0082] Before the intake and at the 12-week visit, participants were asked to fill out a questionnaire about eye symptoms and daily life (DEQS). In the past week, have you experienced any of the following symptoms? "1. Gritty eyes (foreign body sensation), 2. Dry eyes, 3. Painful eyes, 4. Tired eyes, 5. Heavy eyelids, 6. Red eyes." In the past week, have you experienced any of the following? In response to 15 questions, including "7. It is difficult to keep your eyes open, 8. Things appear blurred when you use your eyes, 9. Light is too bright, 10. Eye symptoms get worse when reading newspapers, magazines, books, etc., 11. Eye symptoms get worse when watching TV or using a computer or cell phone, 12. Eye symptoms cause difficulty concentrating, 13. Eye symptoms interfere with work, housework, or studying, 14. You tend to avoid going out because of eye symptoms, and 15. You feel depressed because of eye symptoms," participants were asked to choose "0. Never, 1. Sometimes, 2. Sometimes, 3. Often, 4. Always" in Column A. If they answered "none," they proceeded to the next question. If they answered "any of the above," they were asked to choose "1. Not much of a bother, 2. Somewhat of a bother, 3. A bother, 4. Very of a bother" in Column B. Finally, please tell us about your overall condition, including your eye symptoms over the past week and any associated problems in daily life. Participants were asked to respond with "1. Excellent, 2. Very good, 3. Good, 4. Not very good, 5. Not good, 6. Not good at all." A summary score (total of severity scores for all questions x 25 / number of valid responses) was calculated from the severity scores of 1 to 4 for each question. Eye symptoms were calculated from questions 1 to 6, and impact on daily life scores were calculated from questions 7 to 15. The change in each sub-item and summary score over 12 weeks was calculated by subtracting the pre-intake measurement value from the 12-week post-intake measurement value.

[0083] 3) BUT test (tear film break-up time) Before intake and at the 12-week visit, the fluorescent dye fluorescein was instilled into the eyes, and the time until the breakdown of the tear film appeared was measured as the tear film break-up time (BUT). Average measurements were calculated for each eye from three measurements for the left and right eyes. Furthermore, the left and right average values ​​were calculated from the average measurements for the left and right eyes. For the left and right average values, the 12-week change was calculated by subtracting the pre-intake measurement value from the 12-week post-intake measurement value, and the 12-week change rate was calculated by dividing the pre-intake measurement value from the 12-week post-intake measurement value.

[0084] 4) Schirmer test Before intake and at the 12-week visit, a special test paper was placed between the lower eyelids and left for 5 minutes, and the amount of tear secretion was measured based on the length of time the test paper was wetted with tears. Average measurements were calculated for both the left and right eyes. The 12-week change was calculated by subtracting the pre-intake measurement from the 12-week post-intake measurement, and the 12-week change rate was calculated by subtracting the pre-intake measurement from the 12-week post-intake measurement.

[0085] B. Secondary Evaluation Items 1) Near-point Accommodation Before intake and at the 12-week visit, near-point accommodation was measured using a binocular open-angle constant refraction near-point meter "D'ACOMO." Accommodation was measured according to the measurement method (procedure) of D'ACOMO (WAC Corporation), and accommodation was calculated as 100 ÷ D'ACOMO measurement cm = accommodation. The minimum value of three measurements for each eye was used as the representative measurement. For the representative measurements of the dominant eye, non-dominant eye, left eye, and right eye, the 12-week change was calculated by subtracting the pre-intake measurement from the 12-week post-intake measurement, and the 12-week change rate was calculated by dividing the pre-intake measurement from the 12-week post-intake measurement.

[0086] 2) Computer Vision Syndrome (CVS, Digital Eye Strain Survey) Score Eye strain was assessed before and at the 12-week visit using a digital eye strain survey (Blehm C, Vishnu S, Khattak A, Mitra S, Yee RW. Computer vision syndrome: A review. Surv. Ophthalmol. 2005; 50: 253-262). The change over 12 weeks was calculated by subtracting the pre-insertion measurement from the 12-week post-insertion measurement for each question.

[0087] 3) Subjective symptom questionnaire (stress and fatigue) Before intake and at the 12-week visit, stress and fatigue states were investigated using a stress assessment questionnaire (PHRFSCL-SF) (Imazu Yoshie, Matsuno Toshio, Murakami Masato, Hayashi Yoko, Yamata. Examining the construct validity of the Public Health Research Foundation Stress Checklist Short Form - The relationship between stress reactions and egograms (SGE) in psychosomatic medicine patients - Jpn J Psychosom Med 2016; 56: 263-270). "1. Sudden shortness of breath, 2. Heart palpitations, 3. Sometimes feel dizzy, 4. Sometimes have chest pain, 5. Don't feel like eating even your favorite foods, 6. Have trouble falling asleep and not being able to fall asleep, 7. Feel tired and not able to get rid of fatigue, 8. Sometimes have stiff shoulders and neck, 9. Sometimes have back or waist pain, 10. Eyes get tired easily, 11. Get tired easily when doing something, 12. Have a dull head (feels heavy), 13. Can't do work with confidence, 14. When doing something, feel anxious that it won't go well, 15. Have difficulty approaching things positively." Respondents were asked to answer 24 questions: "I can't do it," "I hesitate and can't make a decision," "I can't make a decision," "I can't make a decision," "I can't make a decision," "I can't handle changes in my environment and keep working," "I feel pressured by the weight of my work," "I sometimes can't trust people," "I sometimes have trouble getting along with people wherever I am," "I want someone who will properly appreciate my efforts," "I sometimes get angry or irritated over small things," "I sometimes lose hope in the future," and "I sometimes feel bad." Responses to questions 1 through 6 were summed to calculate an autonomic nervous system symptom score. Responses to questions 7 through 12 were summed to calculate a fatigue physical response score. Responses to questions 13 through 18 were summed to calculate an anxiety and uncertainty score. Responses to questions 19 through 24 were summed to calculate a depression and mood dysphoria score. For each score, the change over 12 weeks was calculated by subtracting the pre-intake measurement value from the 12-week post-intake measurement value.

[0088] 4) Subjective symptom questionnaire (chilliness, skin condition) Chilliness and skin condition were investigated using VAS and multiple choice questions before intake and at the 12-week visit. For each question, the 12-week change was calculated by subtracting the pre-intake measurement from the 12-week post-intake measurement.

[0089] 5) Blood Total Cholesterol, HDL-C, LDL-C, and Cortisol Blood total cholesterol, HDL-C, LDL-C, and cortisol were measured at the SCR visit and the 12-week visit.

[0090] C. Plasma Lipid Measurement 1) Plasma Sample Preparation Blood was collected from the subject's antecubital vein using EDTA vacutainer tubes. The collected blood was immediately mixed with 20x CTAD solution (a mixture of citric acid, theophylline, adenosine, and dipyridamole). After centrifugation in a refrigerated centrifuge (2500 G x 10 minutes, 4°C), an ATX inhibitor (ONO-8430506, final concentration 10 μM) was added to the plasma. All procedures were performed on ice.

[0091] 2) LC-MS / MS Measurements: Plasma was mixed with a 9-fold volume of methanol containing an internal standard (PC24:0, final concentration 5 μM) and sonicated. After centrifugation in a refrigerated centrifuge (20,000 x g, 5 minutes, 4°C), the supernatant was filtered (YMC Duo-Filter) and used as the LC-MS sample. LC-MS / MS was performed using a NEXERA SERIES LC-40A (Shimadzu Corporation) and a Triple Quad 5500+ (AB Sciex Corporation) according to a previously reported method (Kano K, Matsumoto H, Kono N, Kurano M, Yatomi Y, Aoki J. Suppressing postcollection lysophosphatidic acid metabolism improves the precision of plasma LPA quantification. J Lipid Res. 2021;62:100029). LC separation was performed using a reversed-phase column (C8 CAPCELL PAK UG120 column, 1.5 × 250 mm, Osaka Soda) and an acetonitrile-based mobile phase. MS / MS analysis targeting PC was performed in positive mode, detecting [M+H]+ as the precursor ion and m / z 184 (phosphocholine) as the fragment ion. The peak area of ​​each PC molecular species was calculated from the results, and the area ratio between this and the peak area of ​​the internal standard was used for statistical analysis.

[0092] D. Safety Evaluation For the purpose of safety evaluation, hematology tests (white blood cell count, red blood cell count, hemoglobin content, hematocrit, MCV, MCH, MCHC, platelet count), general biochemistry tests (total protein (TP), albumin (ALB), total bilirubin (T-Bil), ALP, LD (LDH), AST, ALT, γ-GT, CK, triglycerides, urea nitrogen, creatinine, uric acid (UA), blood glucose (fasting), HbA1c (NGSP)), physical examination (body weight, BMI, vital signs), and adverse event investigation were conducted.

[0093] 6. Statistical Analysis R (version 4.2.1) was used on macOS to perform this statistical analysis. The test methods for each evaluation item are described below. Summary statistics were calculated for each evaluation item, and a significance probability (p-value) of less than 0.05 in a two-sided test was considered statistically significant. No adjustment was made for multiplicity of tests associated with multiple hypotheses.

[0094] 1) J-OSDI, DEQS The effects of the test food on each sub-item score (J-OSDI, DEQS), total score (J-OSDI), and summary score (DEQS) were evaluated using analysis of covariance (ANCOVA), with the change 12 weeks after intake as the dependent variable and the pre-intake measurement values ​​as the covariates.

[0095] 2) BUT test, Schirmer test The effects of the test food were evaluated using ANCOVA, with the amount or rate of change 12 weeks after intake as the dependent variable and the pre-intake measurement value as the covariate.

[0096] 3) Near-point accommodation The effect of the test food was evaluated using ANCOVA, with the change in 12 weeks after intake as the objective variable and the pre-intake measurement value as the covariate.

[0097] 4) CVS score The effect of the test food was evaluated using the Brunner-Munzel test for the change in each question 12 weeks after intake.

[0098] 5) Subjective Symptom Questionnaire (Stress / Fatigue) The effect of the test food on the Stress / Fatigue Status Questionnaire (PHRFSCL-SF) was evaluated using ANCOVA with the change in score 12 weeks after intake as the objective variable and the pre-intake measurement value as the covariate. The change in raw score 12 weeks after intake was evaluated using the Brunner-Munzel test.

[0099] 6) Questionnaire on subjective symptoms (chilliness, skin condition) The effects of the study food were evaluated using the Brunner-Munzel test for the values ​​measured before intake and 12 weeks after intake for each question in the questionnaire on chilliness and skin condition.

[0100] 7) Total cholesterol, HDL-C, LDL-C, and blood cortisol The effects of the test food were evaluated using ANCOVA, with the change in 12 weeks after intake as the dependent variable and the pre-intake measurements as the covariate.

[0101] 8) Lipid Measurement The effects of the test food were evaluated using ANCOVA with the change in lipid profile 12 weeks after intake as the dependent variable and the pre-intake measurement values ​​as the covariate.

[0102] 9) Testing methods for safety evaluation items The effects of the test food were evaluated using Welch's t-test or Brunner-Munzel test depending on the normality of the data for the blood test items (hematology tests, general blood biochemistry tests) and physical examinations (body weight, BMI, vital signs) measured before intake and 12 weeks after intake.

[0103] 10) Adverse events, side effects, and other accompanying symptoms were not tested. 11) Subject background For each pre-ingestion measurement, differences between groups were assessed using the chi-square test (without continuity correction) for qualitative variables, and Welch's t-test or Brunner-Munzel test for continuous variables, depending on the normality of the data.

[0104] Results 1. Analysis Population The analysis population for efficacy evaluation consisted of study completers, excluding those who discontinued or dropped out, and excluded subjects who met any of the following analysis exclusion criteria: (1) test food intake less than 90%, (2) significant behavior that compromised the reliability of test results, or (3) failure to meet the inclusion criteria, the exclusion criteria, or failure to comply with the study restrictions. There were no subjects who discontinued or dropped out, or who fell into any of the categories (1) to (3), and all enrolled subjects (44 subjects) were included in the analysis (intention-to-treat analysis, ITT). For safety evaluation, all subjects (44 subjects) who consumed the test food at least once were included in the analysis (Figure 1).

[0105] 2. Primary endpoints The distribution of pre-intake measurement values ​​for the primary endpoints was uniform across all groups compared to the overall population (Table 6).

[0106] 1) J-OSDI and DEQS: The total score, which is the combined score of questions regarding ocular symptoms, vision-related function, and environmental factors, showed a significant decrease in the test food group compared to the placebo group, indicating an overall improvement in subjective eye symptoms. Among the sub-items of ocular symptom score, vision-related function score, and environmental factor score, the ocular symptom score and environmental factor score significantly decreased and improved in the test food group compared to the placebo group. The results are shown in Table 7 and Figure 2A).

[0107]

[0108] Although there was no significant difference in the summary score of the change in DEQS score 12 weeks after ingestion between the placebo and test food groups, there was a tendency for a decrease (p = 0.06). Furthermore, among the sub-items of dry eye symptoms and impact on daily life, dry eye symptoms tended to decrease in the test food group compared to the placebo group (p = 0.07). The results are shown in Table 8 and Figure 2B.

[0109]

[0110] 2) BUT test, Schirmer test No intergroup differences were observed in the left and right average values ​​of BUT test values, either in the amount of change or the rate of change 12 weeks after intake.No intergroup differences were observed in the left and right average values ​​of Schirmer test values, either in the amount of change or the rate of change 12 weeks after intake (Table 9).

[0111]

[0112] 3. Secondary Evaluation Items 1) Near-point Accommodation No differences were observed between groups in the amount of change or rate of change in near-point accommodation 12 weeks after intake. Furthermore, when measuring near-point accommodation, data was missing for 3 of the 44 subjects (6.8%) at the 12-week measurement after intake. This missing data was due to the subject's near-point accommodation falling below the lower limit of the measurement range of the measuring device, and all 3 subjects were in the placebo intake group.

[0113] 2) CVS (Computer Vision Syndrome) score Regarding the change in CVS score 12 weeks after intake, scores for 11 of the 14 items were lower in the test food intake group compared to the placebo intake group, but no differences were observed between the groups in any of the items.

[0114] 3) Subjective Symptom Questionnaire (Stress / Fatigue) In the stress assessment questionnaire (PHRFSCL-SF), among the autonomic nervous system symptom score, fatigue physical response score, anxiety / uncertainty score, and depression / dysphoria score, the fatigue physical response score was significantly reduced and improved in the test food intake group compared to the placebo intake group (Table 10).

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[0116] 4) Subjective Symptom Questionnaire (Chilliness, Skin Condition) No between-group differences were observed in the amount of change in any of the questions in the chilliness questionnaire 12 weeks after intake.No between-group differences were observed in the amount of change in any of the questions in the skin questionnaire 12 weeks after intake.

[0117] 5) Total cholesterol, HDL-C, LDL-C, and blood cortisol No intergroup differences were observed in any of the measured values ​​of total cholesterol, HDL-C, LDL-C, and blood cortisol.

[0118] 4. Plasma lipid measurements In this study, DHA-PC (38:6-PC, 40:6-PC, 40:7-PC, and 44:12-PC) present in plasma was measured using liquid chromatography-mass spectrometry (LC-MS / MS) before and after 12 weeks of placebo or test food intake, and the changes were calculated. The total composition ratio of each molecular species of DHA-PC in plasma was significantly higher in the test food group compared to the placebo group at 12 weeks after intake (Figure 3).

[0119] 5. Safety Evaluation and Adverse Events No differences were observed between groups in hematology tests, general blood biochemistry tests, vital signs, or physical examinations, and the fluctuations were within physiologically normal ranges, so the principal investigator determined that there were no safety issues. The principal investigator determined that the causal relationship of adverse events to test food intake was unrelated or probably unrelated. Commonly occurring events included diarrhea (4 / 22 cases (18.2%) in the placebo group; 3 / 22 cases (13.6%) in the test food group), abdominal pain (1 / 22 cases (4.5%) in the placebo group; 3 / 22 cases (13.6%) in the test food group), and runny nose / nasal congestion (0 / 22 cases (0%) in the placebo group; 3 / 22 cases (13.6%) in the test food group).

[0120] <Safety evaluation through overdose testing> 1. Subjects The purpose of this test was to confirm the safety of excessive intake (5x dose) of salmon roe oil and astaxanthin-containing food "ONO-SR / AST." When examining food safety, it is desirable to conduct and study across a wide range of age groups and genders, so the 5x dose group consisted of 20 subjects to ensure as equal a representation of men and women across all age groups in their 20s to 60s. For reference, to confirm safety at 1x and 2x doses, the number of subjects was reduced to 5 subjects each for the 1x and 2x dose groups.

[0121] 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 pre-examination (lifestyle questionnaire, medical interview, anthropometry, physical examination, and fasting clinical test). Based on the results of the pre-examination, subjects who did not meet the following exclusion criteria were selected.

[0122] Exclusion criteria: (1) individuals who regularly consume health foods rich in DHA, EPA, or astaxanthin; (2) individuals with or a history of serious illnesses such as diabetes, liver disease, kidney disease, or heart disease; (3) individuals at risk of developing allergies related to the study; (4) individuals with illnesses currently being treated that may affect the study, or individuals with a history of chronic or serious illnesses 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 from the pre-examination; (7) individuals who have participated in other clinical trials within one month of obtaining consent to participate in this study, or individuals who plan to participate in other clinical trials 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 their responses to the lifestyle questionnaire; (10) individuals who are otherwise deemed unsuitable as subjects by the responsible investigator.

[0123] 2. Test Food The test food "ONO-SR / AST" (Ono Pharmaceutical Co., Ltd.) was a soft capsule containing 1g of salmon roe oil (150mg DHA, 60mg EPA), 200μg astaxanthin per bag (4 capsules), with gelatin, glycerin, water, and caramel coloring as secondary ingredients. The raw materials and nutritional components are shown in the table below.

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[0125] 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) per day with water or lukewarm water after a meal, without chewing. The intake time 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 continuing up until that point.

[0126] The subjects visited the hospital in the second and fourth weeks of taking the test food, where they underwent a medical interview (to check their physical condition), physical measurements, physical examinations, and fasting clinical tests. They 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), medication use, and other information. Similar tests were planned to be conducted two weeks 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 in the second week after the end of intake to ensure their safety. After the end of the intake period, subjects continued to keep a diary for two weeks.

[0127] 4. Test items and evaluation items Tests included a medical interview to check physical condition and the presence or absence of adverse events, physical measurements including height (pre-test only), weight, and BMI, physical examination including systolic and diastolic blood pressure and pulse rate, hematological tests in fasting clinical tests including white blood cell count, red blood cell count, hemoglobin, hematocrit, and platelet count, blood biochemistry tests including 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, and HbA1c (pre-test only), and urine tests including protein qualitative, glucose quantitative, and occult blood reaction. Except for items that were only required for the pre-test, each test was conducted at the pre-test, week 2, and week 4 of intake. The clinical testing was outsourced to LSI Medience Corporation (Shinagawa-ku, Tokyo).

[0128] The evaluation items were adverse events, measurement values, and test values. Adverse events were subjective symptoms reported in the interview and diary, as well as abnormal fluctuations in test values.

[0129] 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.

[0130] Results 1. Subject Background In this study, a pre-test was conducted on 62 subjects who provided 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. Subject background is shown in Table 2. 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 the study.

[0131] All 30 subjects began taking the test food; however, one male subject dropped out on the 15th day of intake, and one female subject dropped out on the first day after the end of intake due to self-restraint from going out due to the spread of COVID-19. One male subject also dropped out on the first day after the end of intake due to a busy schedule that made it difficult to continue the study. With these three subjects dropping out, 27 subjects (5 in the 1x intake group, 5 in the 2x intake group, and 17 in the 5x intake group) completed the test food intake. Although testing was planned for the second week after the end of intake, due to the COVID-19 state of emergency declaration, subjects were not required to visit the hospital to ensure their safety. Therefore, in this study, safety was evaluated using measurement and test values ​​up to the fourth week. Therefore, measurement and test values ​​were evaluated for the 27 subjects who completed intake, and adverse events were evaluated for all 30 subjects.

[0132] 2. Adverse Events There were 13 cases of fluctuations in measurement values ​​and test values ​​for each subject in 9 subjects. None of the fluctuations in test values ​​were judged to be abnormal (not adverse events). The frequency and total number of adverse events were 0 cases in 0 out of 5 subjects (0%) for the 1x dose, 5 cases in 2 out of 5 subjects (40%) for the 2x dose, and 8 cases in 5 out of 20 subjects (25%) for the 5x dose. 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.

[0133] 3. Measurements and test results 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 doctor in charge of the study determined that all of these changes were at a level that would not pose a clinical problem. Several subjects showed positive clinical test results (urine tests), but this was thought to be due to the effects of menstruation or temporary fluctuations, so the doctor in charge of the study determined that there were no clinical problems.

[0134] Conclusion: The above results indicate that there are no safety issues when an overdose test is conducted in which participants take the salmon roe oil and astaxanthin-containing food "ONO-SR / AST" in amounts of 1, 2, or 5 times the recommended daily intake for four consecutive weeks.

[0135] Discussion The subjective symptoms caused by dry eyes vary widely from person to person, and include not only the simple sensation of dry eyes, but also vague complaints such as eye fatigue, eye grittiness, stinging (pain), and glare. Symptoms such as blurred vision and difficulty seeing can also occur. These eye-related subjective symptoms are said to have a negative impact on the overall quality of life (QOL). Dry eye patients also experience a wide variety of subjective symptoms, including a foreign body sensation, blurred vision, photophobia, eye fatigue, heavy eye pain, and itchy eyes, in addition to dryness.

[0136] The involvement of corneal nociceptors and the sensory nervous system in the mechanism of dry eye has been a hot topic. In 2017, the Tear Film & Ocular Surface Society (TFOS) Dry Eye Workshop (DEWS) II updated the definition of dry eye and added the term "neurosensory abnormalities." The 2016 revision of the Japanese definition and diagnostic criteria for dry eye also reaffirmed the importance of subjective symptoms. It also noted a discrepancy between objective findings and subjective symptoms, with corneal nociceptors and the sensory nervous system being implicated as one of the reasons for this. For example, there are occasional cases in which patients complain of severe pain despite no abnormalities in the corneal and conjunctival epithelium or tear film, and in such cases, a link to neuropathic pain has been suggested.

[0137] In such cases, it is believed that the nerves on the ocular surface or the central nervous system are in a sensitive state, and therapeutic approaches from the sensory system are also attracting attention. When the ophthalmic nerve, which controls corneal sensation in the corneal epithelial cell layer, receives various stimuli, the stimuli are transmitted to the central trigeminal nerve via the trigeminal ganglion. At the end of the ophthalmic nerve, polymodal nociceptors that respond to mechanical, chemical, and thermal stimuli contain receptors and channels such as transient receptor potential ankyrin 1 (TRPA1), transient receptor potential V1 (TRPV1), and acid-sensing ion channel (ASIC), and these responses are thought to cause discomfort and pain. As stated in the J-OSDI question, the discomfort caused by dryness in environments such as strong winds, low humidity, and air-conditioned environments is thought to be due to the above-mentioned sensory nerve sensitivity. It has been reported that nerve block with botulinum toxin, a drug used to treat diabetic neuropathic pain, in patients with neurosensory dry eye reduces photophobia and dryness, regardless of tear film parameters. This suggests that suppression of ophthalmic nerve abnormalities improves subjective symptoms.

[0138] In recent years, a relationship between subjective symptoms of dry eye and psychiatric symptoms has been reported. Patients with depression and anxiety symptoms had higher scores on the Dry Eye Questionnaire (DEQ), a questionnaire about subjective symptoms of dry eye, whereas the results of objective findings of dry eye, such as tear film breakup time (BUT), Schirmer test, and corneal fluorescein staining (FL), were not different from those of healthy individuals. A correlation was also found between depression / anxiety scores and subjective dry eye symptoms. It is presumed that ingestion of the test substance in this study acted on the nervous system, which was in a state of sensitivity to various stimuli, and showed improvement in subjective symptoms and an anti-stress effect without changing objective findings.

[0139] No safety issues were identified in this clinical trial. In addition, no adverse effects were observed in an overdose test (five times the normal dose) of the test food, demonstrating its safety.

[0140] The present disclosure relates to the improvement of eye disorders and can be used in the fields of medicine and food.

Claims

1. A composition for improving eye disorders comprising a fish egg lipid preparation that is formulated to provide a daily intake of about 100 mg to 300 mg of docosahexaenoic acid (DHA).

2. The composition according to claim 1, wherein the fish egg lipid preparation comprises phosphatidylcholine having at least one DHA bound thereto as a constituent fatty acid (DHA-bound PC).

3. The composition of claim 2, wherein the DHA-bound PC comprises palmitoyl-docosahexanoyl-glycerophosphocholine, stearoyl-docosahexanoyl-glycerophosphocholine, oleoyl-docosahexanoyl-glycerophosphocholine, didocosahexanoyl-glycerophosphocholine, or a combination thereof.

4. A composition according to any one of claims 1 to 3, wherein the fish egg lipid preparation contains eicosapentaenoic acid (EPA).

5. The composition according to any one of claims 1 to 4, which is formulated to provide a daily intake of about 100 mg to 300 mg of DHA, about 30 mg to 300 mg of EPA, and about 100 µg to 1000 µg of astaxanthin.

6. The composition according to any one of claims 1 to 5, which is formulated so that about 50 mg to 150 mg of DHA-bound PC is ingested daily.

7. The composition according to any one of claims 1 to 6, which is formulated to provide a daily intake of about 150 mg of DHA, about 60 mg of EPA, and about 200 µg of astaxanthin.

8. The composition according to any one of claims 1 to 7, which is formulated so that about 80 mg of DHA-bound PC is ingested per day.

9. The composition according to any one of claims 1 to 8, wherein the eye complaint is dry eye, foreign body sensation, blurred vision, photophobia, eye fatigue, heavy eye, eye pain or eye itching.

10. The composition according to any one of claims 1 to 9, wherein the eye complaint is dry eye or eye fatigue.

11. The composition according to any one of claims 1 to 10, which further relieves physical stress caused by eye disorders.

12. The composition according to any one of claims 1 to 11, which is taken by a subject aged 40 years or older and younger than 70 years.

13. The composition according to any one of claims 1 to 12, which is taken in one to four divided doses per day.

14. The composition according to any one of claims 1 to 13, which is taken once a day.

15. The composition of any one of claims 1 to 14, wherein the composition is taken daily for at least 12 weeks.

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

17. The composition according to any one of claims 1 to 15, which is a food composition.

18. A composition according to any one of claims 1 to 17 for improving eye disorders through improvement of sensory nerves.

19. A composition for improving ocular sensory nerves, comprising a fish egg lipid preparation that is formulated to provide a daily intake of about 100 mg to 300 mg of docosahexaenoic acid (DHA).

20. The composition of claim 19, wherein the fish egg lipid preparation comprises phosphatidylcholine having at least one DHA bound thereto as a constituent fatty acid (DHA-bound PC).

21. The composition of claim 20, wherein the DHA-bound PC comprises palmitoyl-docosahexanoyl-glycerophosphocholine, stearoyl-docosahexanoyl-glycerophosphocholine, oleoyl-docosahexanoyl-glycerophosphocholine, didocosahexanoyl-glycerophosphocholine, or a combination thereof.

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