Composition for protecting cerebral nerve cells

Abstract

The invention aims to provide a composition for protecting cerebral nerve cells. The present invention relates to a composition for protecting cerebral nerve cells, which is characterized by containing the following components (A) and (B): (A) lutein and / or a fatty acid ester thereof; (B) a docosahexaenoic acid compound, wherein the weight ratio ((B) / (A)) of the component (B) (in terms of docosahexaenoic acid) to the component (A) (in terms of lutein) is 10-120.

Description

Technical Field

[0001] The present invention relates to a composition for protecting brain nerve cells and the like. Background Art

[0002] The brain is a metabolically active tissue with a high oxygen consumption, and at the same time, a large amount of reactive oxygen species is generated. Along with aging, the reactive oxygen species in the brain increase, and many studies have shown that there is a connection between the increase in oxidative stress and the decrease in cognitive function associated with aging. Therefore, it is very important to appropriately control oxidative stress (Non-Patent Document 1, Non-Patent Document 2). The degeneration or necrosis of brain nerve cells caused by aging or oxidative stress can trigger diseases or disorders such as dementia or the decline of higher brain functions and cognitive functions. Therefore, a highly safe method for protecting brain nerve cells is desired.

[0003] Carotenoids are antioxidants produced by plants, algae, bacteria, etc., and are rich in vegetables or fruits, and their powerful antioxidant effects are known (Non-Patent Document 3, Non-Patent Document 4). It has been reported that 16 types of carotenoids are present in the human brain (Non-Patent Document 5), and among them, lutein and zeaxanthin, which are a type of carotenoids rich in Komatsuna etc. (Non-Patent Document 6), have a nerve cell protection effect based on the antioxidant effect in the brain nervous system (Non-Patent Document 7, Non-Patent Document 8). In addition, docosahexaenoic acid (hereinafter also referred to as DHA), which is a fatty acid component of the oil contained in blue fish such as mackerel, saury, and sardines, is also known to have various physiological activities. Non-Patent Document 8 reports the nerve cell protection effect of a composition containing lutein and DHA and having a weight ratio of DHA to lutein of 0.72, but its effect is equivalent to the effect of DHA alone. Patent Document 1 discloses an oral composition for improving brain function containing DHA, capsaicin, lutein, and zeaxanthin.

[0004] Patent Documents Patent Document 1: Japanese Patent No. 6375296 Gazette

[0005] Non-Patent Documents Non-Patent Document 1: Ionescu-Tucker, A. & Cotman, C.W. Neurobiol. Aging 107, 86 - 95 (2021) Non-Patent Document 2: Kandlur, A. et al. Front. Mol. Neurosci. 13, 41 (2020) Non-Patent Document 3: Manochkumar, J. et al. Phytomedicine 91, 153676 (2021) Non-Patent Document 4: Maoka, T. J. Nat. Med. 74, 1-16 (2020) Non-Patent Document 5: Craft, N. E. et al. J. Nutr. Health Aging 8, 156-162 (2004) Non-Patent Document 6: Aizawa, K. & Inakuma, T. Food Sci. Technol. Res. 13, 247-252 (2007) Non-Patent Document 7: Singhrang, N. et al. J. Biomed. Pharmacother. 103, 582-587 (2018) Non-Patent Document 8: Hu, Y. et al. J. Nutr. Sci. Vitaminol. (Tokyo). 67, 234-242 (2021) SUMMARY OF THE INVENTION

[0006] In Patent Document 1, the protective effect on brain nerve cells based on the above-mentioned oral composition is not clear.

[0007] An object of the present invention is to provide a composition for protecting brain nerve cells.

[0008] The present inventors conducted in-depth research to solve the above problems and found that by combining lutein and / or its fatty acid esters and docosahexaenoic acid compounds within a specific ratio range, compared with using only lutein and / or its fatty acid esters or only docosahexaenoic acid compounds, brain nerve cells can be protected more effectively (synergistically), thus completing the present invention.

[0009] The present invention relates to the following composition. [1] A composition for protecting brain nerve cells, characterized by containing the following components (A) and (B): (A) Lutein and / or its fatty acid esters; (B) Docosahexaenoic acid compounds, and the weight ratio of component (B) (docosahexaenoic acid conversion) to component (A) (lutein conversion), i.e., (B) / (A), is 10 to 120. [2] The composition according to the above [1], characterized in that the docosahexaenoic acid compound is at least one selected from free docosahexaenoic acid, its salts, and compounds containing docosahexaenoic acid as a constituent fatty acid. [3] The composition according to [2] above is characterized in that the compound containing docosahexaenoic acid as a constituent fatty acid is selected from triglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid, diglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid, monoglycerides in which the constituent fatty acid is docosahexaenoic acid, phospholipids in which part or all of the constituent fatty acids are docosahexaenoic acid, glycolipids in which part or all of the constituent fatty acids are docosahexaenoic acid, and alcohol esters of docosahexaenoic acid. [4] The composition according to any one of [1] to [3] above is characterized in that component (B) is derived from marine products. [5] The composition according to any one of [1] to [4] above is characterized in that it is an oral composition. [6] The composition according to any one of [1] to [5] above is characterized in that it is a food or drink. [7] The composition according to any one of [1] to [6] above is characterized in that it is labeled with at least one function selected from "maintaining cognitive function", "maintaining memory function", "maintaining attention function", "inhibiting brain nerve cell disorders", "protecting brain nerve cells", "guarding brain nerve cells", "reducing brain oxidative stress", "guarding the brain from oxidative stress", and "antioxidation of the brain". [8] The composition according to any one of [1] to [7] above is characterized in that it further contains zeaxanthin and / or its fatty acid ester, arachidonic acid compounds, and eicosapentaenoic acid compounds. [9] The composition according to [8] is characterized in that it is used for improving memory function or inhibiting its decline.

[0010] According to the present invention, a composition for protecting brain nerve cells can be provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 It is a graph showing the change rate of the LDH release level based on the composition containing lutein and DHA (Examples 1 to 4, Comparative Example 6) and the predicted values of the change rate (Comparative Example 2 + Comparative Example 5, Comparative Example 3 + Comparative Example 5, Comparative Example 2 + Comparative Example 4, and Comparative Example 3 + Comparative Example 4). DETAILED DESCRIPTION

[0012] The composition for protecting brain nerve cells of the present invention (hereinafter also referred to as "the composition of the present invention") contains the following components (A) and (B): (A) lutein and / or its fatty acid ester; (B) docosahexaenoic acid compounds, and the weight ratio of component (B) (docosahexaenoic acid conversion) to component (A) (lutein conversion) ((B) / (A)) is 10 to 120. The composition of the present invention contains lutein and / or its fatty acid esters and docosahexaenoic acid compounds as active ingredients.

[0013] (Component (A): Lutein and / or its fatty acid esters) The composition of the present invention contains lutein and / or its fatty acid esters. Lutein is a kind of carotenoid. Lutein and / or its fatty acid esters are contained in plants, etc., and can be prepared by extraction from plants, for example. Fatty acid esters of lutein are contained in green and yellow vegetables such as spinach, kale, broccoli, pumpkin, carrot, and green pepper; fruits such as orange, peach, papaya, plum, mango, avocado, raspberry, and rosehip; petals of marigold, etc. In the present invention, as long as the effects of the present invention can be exerted, plant-derived raw materials rich in lutein and / or its fatty acid esters can also be contained in the composition of the present invention. Lutein and / or its fatty acid esters can also use commercially available products.

[0014] The fatty acid ester of lutein is a fatty acid ester in which one or two hydroxyl groups of lutein are bonded to fatty acids. As the fatty acid in the fatty acid ester of lutein, fatty acids having 4 to 20 carbon atoms can be cited, and fatty acids having 8 to 18 carbon atoms are preferably cited. The fatty acid ester of lutein contained in the composition can be one kind or two or more kinds. Lutein is generated by hydrolyzing the fatty acid ester of lutein.

[0015] (Component (B): Docosahexaenoic acid compounds) The composition of the present invention contains docosahexaenoic acid compounds. As the docosahexaenoic acid compounds, docosahexaenoic acid (free DHA), its salts, and their derivatives can be used. As derivatives of docosahexaenoic acid, for example, compounds having a structure in which docosahexaenoic acid is bonded and compounds that can separate docosahexaenoic acid by hydrolysis can be used. As the compounds that can separate docosahexaenoic acid by hydrolysis, compounds containing docosahexaenoic acid as a constituent fatty acid can be cited. The docosahexaenoic acid compounds contained in the composition can be one kind or two or more kinds.

[0016] The docosahexaenoic acid compounds used in the present invention are preferably selected from free docosahexaenoic acid, its salts, and compounds containing docosahexaenoic acid as a constituent fatty acid. As salts of docosahexaenoic acid, for example, calcium salts, sodium salts, etc. can be cited.

[0017] As a compound containing docosahexaenoic acid as a constituent fatty acid, it is preferably a compound selected from triglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid, diglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid, monoglycerides in which the constituent fatty acid is docosahexaenoic acid, phospholipids in which part or all of the constituent fatty acids are docosahexaenoic acid, glycolipids in which part or all of the constituent fatty acids are docosahexaenoic acid, and alcohol esters of docosahexaenoic acid. Among them, as docosahexaenoic acid compounds, triglycerides, diglycerides and phospholipids in which part or all of the constituent fatty acids are docosahexaenoic acid are more preferred, triglycerides and diglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid are further preferred, and triglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid are particularly preferred.

[0018] The docosahexaenoic acid compounds used in the present invention are not limited by their form, manufacturing method, etc., and chemically synthesized products can be used, and substances from natural products or substances from genetically recombinant organisms (such as Escherichia coli, yeast, plants, etc.) can also be used. As substances from natural products, substances from marine products or algae, fungi (bacteria of the genus Colwellia, slime molds, etc.), plants (Marchantia polymorpha, etc.) with DHA production ability can be cited. Among them, docosahexaenoic acid compounds are preferably from marine products. In addition, docosahexaenoic acid compounds extracted and / or refined from raw materials such as natural products containing docosahexaenoic acid compounds can also be used. The raw materials can be used directly, or docosahexaenoic acid compounds can be contained in the composition. As raw materials containing docosahexaenoic acid compounds, for example, edible oils such as oils extracted from fish oil, meat, and milk can be cited. As fish oil containing docosahexaenoic acid compounds, fish oil concentrated in such a manner that the content rate of docosahexaenoic acid compounds (in terms of docosahexaenoic acid) reaches 5% by weight or more, preferably 10% by weight or more, and more preferably 50% by weight or more is preferred. For example, docosahexaenoic acid compounds extracted and / or refined from fish oil containing docosahexaenoic acid compounds can be used, or the fish oil can be used directly. In one mode, the composition of the present invention contains fish oil, and the docosahexaenoic acid compound can be a part of the fish oil.

[0019] In the composition of the present invention, the weight ratio ((B) / (A)) of (B) docosahexaenoic acid compounds (in terms of docosahexaenoic acid) to (A) lutein and / or its fatty acid ester (in terms of lutein) is 10 to 120. By combining lutein and / or its fatty acid esters (A) and docosahexaenoic acid compounds (B) in the above weight ratio, excellent effects can be significantly obtained in the protection of brain nerve cells. The weight ratio is preferably 15 to 100, more preferably 20 to 80, and still more preferably 20 to 60. The amount in terms of lutein or a similar expression refers to its amount when component (A) is lutein, and when component (A) is a fatty acid ester of lutein, it refers to the value obtained by multiplying the molar number of the ester by the molecular weight of lutein. The amount of lutein and / or its fatty acid esters can be measured by HPLC (high performance liquid chromatography). The amount in terms of docosahexaenoic acid or a similar expression refers to its amount when component (B) is free DHA. When component (B) is not free DHA, it refers to the amount of free DHA obtained by saponification treatment or enzymatic treatment to free DHA. The amount of free DHA can be measured by gas chromatography (GC). When using two or more components of (A), the amount of component (A) is the total amount thereof. When using two or more components of (B), the amount of component (B) is the total amount thereof.

[0020] In this specification, the so-called brain nerve cells are nerve cells constituting the brain tissue. In addition, the so-called protection of brain nerve cells means inhibiting brain nerve cell disorders or inhibiting brain nerve cell death. The protective effect on brain nerve cells can be evaluated according to the common knowledge in the technical field to which the present invention pertains, and the method is not particularly limited. For example, the protective effect on brain nerve cells can be evaluated by measuring the amount of LDH released outside the brain nerve cells.

[0021] The composition of the present invention can be used for the prevention or improvement of a state or disease that can be expected to be prevented or improved by protecting brain nerve cells. Examples of such a state or disease include a decrease in cognitive function, a decrease in memory function, a decrease in attention function, brain nerve cell death, brain nerve cell disorders, an increase in brain oxidative stress, etc. In one aspect, the composition of the present invention can be used for the prevention or improvement of such a state or disease. In one aspect, the composition of the present invention can also be used for an object in need of preventing or improving such a state or disease. In this specification, the prevention of a state or disease includes: preventing the onset of the state or disease, delaying the onset of the state or disease, reducing the onset rate of the state or disease, reducing the risk of onset of the state or disease, etc. The improvement of a state or disease includes: restoring the object from the state or disease, alleviating the symptoms of the state or disease, delaying or preventing the development of the state or disease, etc. Restoration includes at least partial restoration. The composition of the present invention is applicable to either therapeutic or non-therapeutic uses. The so-called non-therapeutic means not including medical acts, that is, the concepts of human surgery, treatment or diagnosis.

[0022] The composition of the present invention can be provided in forms such as food and drink products, pharmaceuticals, quasi-drugs, feeds, etc., but is not limited to these. The composition of the present invention can itself be a food and drink product, pharmaceutical, quasi-drug, feed, etc. for protecting nerve cells in the brain, or can also be a preparation, material such as an additive used in these. As an example, the composition of the present invention can be provided in the form of a preparation, but is not limited to this form. The preparation can be directly provided as the composition or provided as a composition containing the preparation. The composition of the present invention can also be called a preparation for protecting nerve cells in the brain. The composition of the present invention can be either an oral composition or a non-oral composition, but is preferably an oral composition. According to the present invention, an oral composition having an excellent nerve cell protection effect in the brain can be provided. As the oral composition, food and drink products, oral pharmaceuticals, oral quasi-drugs can be cited, and food and drink products are preferred.

[0023] Within the range not impairing the effects of the present invention, in addition to component (A) and component (B), the composition of the present invention can contain arbitrary additives and arbitrary components. These additives and components can be selected according to the form of the composition, etc., and substances that can be generally used in food and drink products, pharmaceuticals, quasi-drugs, feeds, etc. can be used.

[0024] As arbitrary components, zeaxanthin and / or its fatty acid esters, arachidonic acid compounds, eicosapentaenoic acid compounds, etc. can be cited. The composition of the present invention further preferably contains zeaxanthin and / or its fatty acid esters, arachidonic acid compounds and eicosapentaenoic acid compounds. In one mode, the composition of the present invention can contain lutein and / or its fatty acid esters, docosahexaenoic acid compounds, zeaxanthin and / or its fatty acid esters, arachidonic acid compounds and eicosapentaenoic acid compounds as active ingredients.

[0025] Zeaxanthin is a kind of carotenoid. Zeaxanthin and / or its fatty acid esters are contained in vegetables such as green peppers and spinach, or in eggs, petals of marigolds, etc., and can be prepared, for example, by extraction from plants. The composition of the present invention can also contain plant-derived raw materials rich in zeaxanthin and / or its fatty acid esters, etc. Zeaxanthin and / or its fatty acid esters can also be commercially available products.

[0026] The fatty acid esters of zeaxanthin are fatty acid esters in which one or two hydroxyl groups of zeaxanthin are bonded to fatty acids. Examples of the fatty acids in the fatty acid esters of zeaxanthin include fatty acids having 4 to 20 carbon atoms, preferably fatty acids having 8 to 18 carbon atoms. The fatty acid esters of zeaxanthin contained in the composition may be one kind or two or more kinds. Hydrolysis of the fatty acid esters of zeaxanthin generates zeaxanthin.

[0027] As arachidonic acid compounds, arachidonic acid (free arachidonic acid), its salts and their derivatives can be used. As derivatives of arachidonic acid, for example, compounds having a structure in which arachidonic acid is bonded and compounds that can release arachidonic acid by hydrolysis can be used. As compounds that can release arachidonic acid by hydrolysis, compounds containing arachidonic acid as a constituent fatty acid can be cited. Arachidonic acid compounds can be used alone or in combination of two or more. Arachidonic acid compounds are preferably selected from free arachidonic acid, its salts and compounds containing arachidonic acid as a constituent fatty acid. Examples of the salts of arachidonic acid include calcium salts and sodium salts. As compounds containing arachidonic acid as a constituent fatty acid, preferably selected from triglycerides in which part or all of the constituent fatty acids are arachidonic acid, diglycerides in which part or all of the constituent fatty acids are arachidonic acid, monoglycerides in which the constituent fatty acid is arachidonic acid, phospholipids in which part or all of the constituent fatty acids are arachidonic acid, glycolipids in which part or all of the constituent fatty acids are arachidonic acid, and alcohol esters of arachidonic acid. Among them, as arachidonic acid compounds, more preferably triglycerides, diglycerides and phospholipids in which part or all of the constituent fatty acids are arachidonic acid, still more preferably triglycerides and diglycerides in which part or all of the constituent fatty acids are arachidonic acid, and particularly preferably triglycerides in which part or all of the constituent fatty acids are arachidonic acid.

[0028] Arachidonic acid compounds are not limited by their form, production method, etc., and chemically synthesized products, substances from natural products or substances from genetically recombinant organisms (Escherichia coli, yeast, plants, etc.) can be used. As substances from natural products, substances from marine products, algae, microorganisms, plants (Marchantia polymorpha, etc.) having ARA production ability can be cited. Among them, the arachidonic acid compounds are preferably derived from microorganisms. Examples of the microorganisms include those belonging to the genera Mortierella, Conidiobolus, Pythium, Phytophthora, Penicillium, Cladosporium, Mucor, Fusarium, Aspergillus, Rhodotorula, Entomophthora, Echinosporangium, and Saprolegnia. In addition, arachidonic acid compounds extracted and / or refined from raw materials such as natural substances containing arachidonic acid compounds can be used, or the raw materials can be directly used to incorporate arachidonic acid compounds into the composition. Examples of the raw materials containing arachidonic acid compounds include edible oils and fats such as oils and fats produced by microorganisms, and oils and fats extracted from eggs, meats, and marine products. The production method of arachidonic acid compounds using microorganisms is not limited, and known methods can be used, such as the method described in Japanese Patent Laid-Open No. 2016-172770.

[0029] As the eicosapentaenoic acid compounds, eicosapentaenoic acid (free eicosapentaenoic acid. Hereinafter also referred to as EPA), its salts, and their derivatives can be used. The eicosapentaenoic acid compounds can be used alone or in combination of two or more. Examples of the derivatives of eicosapentaenoic acid include compounds having a structure in which eicosapentaenoic acid is bonded and compounds that can release eicosapentaenoic acid by hydrolysis. Examples of the compounds that can release eicosapentaenoic acid by hydrolysis include compounds containing eicosapentaenoic acid as a constituent fatty acid. The eicosapentaenoic acid compounds can be used alone or in combination of two or more. The eicosapentaenoic acid compounds are preferably selected from free eicosapentaenoic acid, its salts, and compounds containing eicosapentaenoic acid as a constituent fatty acid. Examples of the salts of eicosapentaenoic acid include calcium salts and sodium salts. As the compounds containing eicosapentaenoic acid as a constituent fatty acid, compounds selected from triglycerides in which part or all of the constituent fatty acids are eicosapentaenoic acid, diglycerides in which part or all of the constituent fatty acids are eicosapentaenoic acid, monoglycerides in which the constituent fatty acid is eicosapentaenoic acid, phospholipids in which part or all of the constituent fatty acids are eicosapentaenoic acid, glycolipids in which part or all of the constituent fatty acids are eicosapentaenoic acid, and alcohol esters of eicosapentaenoic acid are preferred. Among them, as eicosapentaenoic acid compounds, triglycerides, diglycerides and phospholipids in which part or all of the constituent fatty acids are eicosapentaenoic acid are more preferred, triglycerides and diglycerides in which part or all of the constituent fatty acids are eicosapentaenoic acid are further preferred, and triglycerides in which part or all of the constituent fatty acids are eicosapentaenoic acid are particularly preferred.

[0030] The eicosapentaenoic acid compounds are not limited by their form, manufacturing method, etc., and chemical synthetic products can be used, or substances from natural products or substances from genetically recombinant organisms (Escherichia coli, yeast, plants, etc.) can be used. As substances from natural products, substances from marine products or algae, fungi (such as slime molds), plants (such as liverworts) with EPA production capacity can be cited. Among them, the eicosapentaenoic acid compounds are preferably from marine products. In addition, eicosapentaenoic acid compounds extracted and / or refined from raw materials such as natural products containing eicosapentaenoic acid compounds can be used. The raw materials can be used directly, or the eicosapentaenoic acid compounds can be contained in the composition. As raw materials containing eicosapentaenoic acid compounds, for example, edible oils and fats such as oils extracted from fish oil, eggs, meat, milk or algae can be cited. As fish oil containing eicosapentaenoic acid compounds, fish oil concentrated in such a way that the content rate of eicosapentaenoic acid compounds (converted to eicosapentaenoic acid) reaches 5% by weight or more, preferably 10% by weight or more, more preferably 20% by weight or more is preferred. For example, eicosapentaenoic acid compounds extracted and / or refined from fish oil containing eicosapentaenoic acid compounds can be used, or the fish oil can be used directly.

[0031] In one mode of the present invention, a composition containing (A) lutein and / or its fatty acid ester, (B) docosahexaenoic acid compounds, zeaxanthin and / or its fatty acid ester, arachidonic acid compounds and eicosapentaenoic acid compounds, and having a weight ratio ((B) / (A)) of component (B) (converted to docosahexaenoic acid) to component (A) (converted to lutein) of 10 to 120 is preferably used for improving cognitive function or suppressing its decline. As the cognitive function, memory function is preferred. Cognitive functions such as memory function can be evaluated, for example, by the Cognitrax test. Cognitrax is a cognitive function test designed for Japanese people based on CNS vital Signs developed by CNSVital Signs, Inc. in the United States (Gualtieri CT, et al. Reliability and validity of a computerized neurocognitive test battery, CNS Vital Signs. Arch Clin Neuropsychol 21(7): 623-643 (2006)). The composition of the present invention can be used to improve, for example, the comprehensive memory evaluated by the Cognitrax test.

[0032] When the composition of the present invention is made into foods or drinks, ingredients that can be used in foods or drinks (for example, food materials, food additives used as needed, etc.) can be formulated into components (A) and (B) to make various foods or drinks. The foods or drinks are not particularly limited, and examples include general foods or drinks, health foods, health drinks, foods with functional claims, foods for specified health use, health supplements, foods for patients, etc. The above-mentioned health foods, foods with functional claims, foods for specified health use, health supplements, etc. can be used in various preparation forms such as fine granules, tablets, granules, powders, capsules, chewable tablets, dry syrups, syrups, liquids, drinks, liquid foods, etc.

[0033] When the composition of the present invention is made into pharmaceuticals or quasi-drugs, pharmacologically acceptable carriers, additives, etc. can be formulated into components (A) and (B) to make pharmaceuticals in various dosage forms. The dosage form can be made into a dosage form suitable for administration. As dosage forms for oral administration, for example, there can be mentioned solid oral preparations for oral administration such as tablets, coated tablets, fine granules, granules, powders, pills, capsules, dry syrups, chewable tablets, etc.; liquid oral preparations for oral administration such as oral liquids, syrups, etc. As dosage forms for non-oral administration, there can be mentioned injections, drip infusions, ointments, lotions, patches, suppositories, nasal preparations, pulmonary preparations (inhalants), etc. The pharmaceuticals can also be pharmaceuticals for non-human animals. As the mode of administration (ingestion) of the pharmaceuticals or quasi-drugs, there can be mentioned modes of oral or non-oral (transdermal, transmucosal, enteral, injection, etc.) administration. From the viewpoint of more fully obtaining the effects of the present invention, the mode of administration of the pharmaceuticals or quasi-drugs is preferably the oral administration mode. The carriers, additives, etc. only need to be pharmacologically acceptable substances that can be used in pharmaceuticals or quasi-drugs. For example, one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, etc. can be mentioned.

[0034] When the composition of the present invention is made into feed, components that can be used in feed can be formulated into the components (A) and (B) to make feed. Examples of the feed include livestock feed for cattle, pigs, chickens, sheep, horses, etc.; small animal feed for rabbits, guinea pigs, rats, mice, etc.; pet foods for dogs, cats, little birds, etc.

[0035] When the composition of the present invention is made into food and drink products, pharmaceuticals, quasi-drugs, feed, etc., its manufacturing method is not particularly limited, and components (A) and (B) can be used and manufactured by general methods.

[0036] The contents of component (A) and component (B) in the composition of the present invention can be appropriately set according to the form of the composition and the like. The total content (in terms of lutein) of lutein and / or its fatty acid esters in the composition of the present invention is, for example, preferably 0.01% by weight or more, more preferably 0.1% by weight or more, and further preferably 9% by weight or less in the composition. In one mode, the total content (in terms of lutein) of lutein and / or its fatty acid esters is preferably 0.01 - 9% by weight, more preferably 0.1 - 9% by weight in the composition.

[0037] The total content (in terms of docosahexaenoic acid) of docosahexaenoic acid compounds in the composition of the present invention is, for example, preferably 1% by weight or more, more preferably 10% by weight or more, and further preferably 80% by weight or less in the composition. In one mode, the total content (in terms of docosahexaenoic acid) of docosahexaenoic acid compounds is preferably 1 - 80% by weight, more preferably 10 - 80% by weight in the composition.

[0038] When the composition of the present invention contains zeaxanthin and / or its fatty acid esters, its total content (in terms of zeaxanthin) is, for example, preferably 0.01% by weight or more, more preferably 0.02% by weight or more, and further preferably 5% by weight or less, more preferably 2% by weight or less in the composition. In one mode, the total content (in terms of zeaxanthin) of zeaxanthin and / or its fatty acid esters is preferably 0.01 - 5% by weight, more preferably 0.02 - 2% by weight in the composition.

[0039] When the composition of the present invention contains arachidonic acid compounds, its total content (in terms of arachidonic acid conversion) is, for example, preferably 2% by weight or more, more preferably 4% by weight or more, further preferably 40% by weight or less, and more preferably 32% by weight or less in the composition. In one mode, the total content (in terms of arachidonic acid conversion) of arachidonic acid compounds is preferably 2 to 40% by weight, more preferably 4 to 32% by weight in the composition. The content of arachidonic acid can be determined by gas chromatography (GC) method.

[0040] When the composition of the present invention contains eicosapentaenoic acid compounds, its total content (in terms of eicosapentaenoic acid conversion) is, for example, preferably 1% by weight or more, more preferably 3% by weight or more, further preferably 40% by weight or less, and more preferably 30% by weight or less in the composition. In one mode, the total content (in terms of eicosapentaenoic acid conversion) of eicosapentaenoic acid compounds is preferably 1 to 40% by weight, more preferably 3 to 30% by weight in the composition. The content of eicosapentaenoic acid can be determined by gas chromatography (GC) method.

[0041] The composition of the present invention can be ingested or administered by an appropriate method according to its form. The composition of the present invention is preferably administered orally or ingested orally. The intake amount (which can also be called the dosage amount) of the composition of the present invention is not particularly limited, and it can be an amount (effective amount) that can obtain the effect of protecting nerve cells in the brain, and can be appropriately set according to the administration form, administration method, body weight of the subject, etc. For example, when the composition of the present invention is orally administered or ingested to humans (adults), the total dosage of lutein and / or its fatty acid esters, in terms of lutein conversion, per 60 kg of body weight per day, is preferably 0.5 mg or more, more preferably 1 mg or more, further preferably 3 mg or more, further preferably 200 mg or less, more preferably 100 mg or less, and further preferably 80 mg or less. In addition, the total dosage of docosahexaenoic acid compounds, in terms of docosahexaenoic acid conversion, per 60 kg of body weight per day, is preferably 10 mg or more, more preferably 100 mg or more, further preferably 2,000 mg or less, and more preferably 1,000 mg or less. In one mode, the total dosage of lutein and / or its fatty acid esters, in terms of lutein conversion, for humans (adults), per 60 kg of body weight per day, is preferably 0.5 to 200 mg, more preferably 1 to 100 mg, and further preferably 3 to 80 mg. In addition, the total dosage of docosahexaenoic acid compounds, in terms of docosahexaenoic acid conversion, for humans (adults), per 60 kg of body weight per day, is preferably 10 to 2,000 mg, more preferably 100 to 2,000 mg, and further preferably 100 to 1,000 mg. Preferably, the above amount is divided into one or more administrations per day, for example, one or more times (e.g., 2 to 3 times) per day for ingestion or administration. In one mode, it is preferred that the above amount of lutein and / or its fatty acid esters and docosahexaenoic acid compounds be orally ingested or administered to humans. In one mode, the composition of the present invention can be used to enable a human (adult) to ingest or administer the above amount of lutein and / or its fatty acid esters and docosahexaenoic acid compounds per 60 kg of body weight per day. In addition, when two or more kinds of lutein and / or its fatty acid esters are used, the total dosage of lutein and / or its fatty acid esters is the total amount thereof. When two or more kinds of docosahexaenoic acid compounds are used, the total dosage of docosahexaenoic acid compounds is the total amount thereof.

[0042] When the composition of the present invention contains zeaxanthin and / or its fatty acid esters, the total dosage of zeaxanthin and / or its fatty acid esters for oral administration or ingestion to humans (adults) is preferably 0.1 mg or more, more preferably 0.5 mg or more, further preferably 50 mg or less, more preferably 30 mg or less, and still further preferably 20 mg or less per 60 kg of body weight per day in terms of zeaxanthin. In one mode, the total dosage of zeaxanthin and / or its fatty acid esters, in terms of zeaxanthin, for humans (adults) is preferably 0.1 to 50 mg, more preferably 0.1 to 30 mg, and still further preferably 0.5 to 20 mg per 60 kg of body weight per day.

[0043] When the composition of the present invention contains arachidonic acid compounds, the total dosage of arachidonic acid compounds for oral administration or ingestion to humans (adults) is preferably 10 mg or more, more preferably 20 mg or more, further preferably 30 mg or more, further preferably 500 mg or less, more preferably 400 mg or less, and still further preferably 300 mg or less per 60 kg of body weight per day in terms of arachidonic acid. In one mode, the total dosage of arachidonic acid compounds, in terms of arachidonic acid, for humans (adults) is preferably 10 to 500 mg, more preferably 20 to 400 mg, and still further preferably 30 to 300 mg per 60 kg of body weight per day.

[0044] When the composition of the present invention contains eicosapentaenoic acid compounds, the total dosage of eicosapentaenoic acid compounds when orally administered or ingested to humans (adults) is, in terms of eicosapentaenoic acid conversion, per 60 kg of body weight per day, preferably 10 mg or more, more preferably 20 mg or more, still more preferably 30 mg or more. In addition, it is preferably 500 mg or less, more preferably 400 mg or less, still more preferably 300 mg or less. In one embodiment, the total dosage of eicosapentaenoic acid compounds, in terms of eicosapentaenoic acid conversion, for humans (adults), is per 60 kg of body weight per day, preferably 10 - 500 mg, more preferably 20 - 400 mg, still more preferably 30 - 300 mg.

[0045] The composition of the present invention is preferably continuously ingested or administered. By continuously ingesting or administering lutein and / or its fatty acid esters and docosahexaenoic acid compounds, it is expected to enhance the above effects. In one embodiment, the composition of the present invention is preferably continuously ingested or administered for 1 week or more, more preferably 4 weeks or more, still more preferably 8 weeks or more, and particularly preferably 12 weeks or more.

[0046] The administration subject is preferably a human or non - human mammal, more preferably a human. In addition, as the administration subject of the composition of the present invention, preferably those in need or desiring to protect nerve cells of the brain are selected. For example, preferred subjects may include those desiring to prevent or improve the above - mentioned conditions or diseases, etc. In one embodiment, the composition of the present invention can also be used for healthy subjects for the purpose of preventing or improving conditions or diseases that can be expected to be prevented or improved by protecting nerve cells of the brain.

[0047] In one embodiment, as the administration subject in the present invention, the elderly and middle - aged are preferably selected. Generally, the elderly refer to humans aged 65 or above, and the middle - aged refer to humans aged 45 or above and below 65. As the administration subject of the composition of the present invention, for example, humans aged 45 or above are preferably selected, and more preferably humans aged 55 or above and below 80. In one embodiment, as the administration subject of the composition of the present invention, more preferably those with reduced cognitive function are selected.

[0048] On the composition of the present invention, one or more of the use, types of active ingredients, the above - mentioned effects, methods of use (e.g., ingestion method, administration method), etc. can be indicated on the packaging, container, instruction manual, etc. The composition of the present invention can also be attached with an indication meaning the protection of nerve cells of the brain or the effects based on these effects. On the composition of the present invention, labels of one or more functions such as "maintaining cognitive function", "maintaining memory function", "maintaining attention function", "inhibiting nerve cell disorders", "protecting nerve cells", "guarding nerve cells", "reducing brain oxidative stress", "guarding the brain from oxidative stress", and "antioxidation of the brain" may also be attached. In one embodiment of the present invention, the composition of the present invention is preferably a food or drink product with the above labels. In addition, the above labels may also be labels indicating the meaning of being used to obtain the above functions.

[0049] The present invention also includes the use of lutein and / or its fatty acid esters and docosahexaenoic acid compounds for manufacturing a composition for protecting nerve cells. In the above use, the weight ratio of docosahexaenoic acid compounds (in terms of docosahexaenoic acid) to lutein and / or its fatty acid esters (in terms of lutein) is 10 to 120.

[0050] The present invention also includes the following method. A method for protecting nerve cells, characterized by comprising administering a composition containing the following components (A) and (B): (A) Lutein and / or its fatty acid esters; (B) Docosahexaenoic acid compounds, and the weight ratio ((B) / (A)) of component (B) (in terms of docosahexaenoic acid) to component (A) (in terms of lutein) is 10 to 120.

[0051] The present invention also includes the following use. A use, characterized by using the following composition for protecting nerve cells. A composition, characterized by containing the following components (A) and (B): (A) Lutein and / or its fatty acid esters; (B) Docosahexaenoic acid compounds, and the weight ratio ((B) / (A)) of component (B) (in terms of docosahexaenoic acid) to component (A) (in terms of lutein) is 10 to 120. The above method or use may be a therapeutic method or use, or a non-therapeutic method or use. By administering the above composition to an object, a protective effect on nerve cells can be obtained.

[0052] In the above use and method, the preferred forms of lutein and / or its fatty acid esters, docosahexaenoic acid compounds, their ratios, etc. are the same as those of the composition of the present invention above. As lutein and / or its fatty acid esters and docosahexaenoic acid compounds, one kind can be used respectively, or two or more kinds can be used. In the above applications and methods, it is preferred to administer (let the subject ingest) the above composition to the subject once or more a day, for example, once to multiple times (for example, 2 to 3 times) a day. In the above applications and methods, it is preferred to orally administer (ingest) the above composition. The above application is preferably for humans or non-human mammals, and more preferably for humans.

[0053] In the above applications and methods, it is sufficient to use one or more kinds of lutein and / or its fatty acid esters and one or more kinds of docosahexaenoic acid compounds in an amount (which can also be called an effective amount) that can achieve the desired effect. The preferred dosage or the subject to be administered of lutein and / or its fatty acid esters and docosahexaenoic acid compounds is the same as that of the composition of the present invention above.

[0054] In this specification, a numerical range represented by a lower limit value and an upper limit value, that is, "lower limit value to upper limit value" includes these lower limit values and upper limit values. For example, the range represented by "1 to 2" means 1 or more and 2 or less, and includes 1 and 2. In this specification, the upper limit and the lower limit can be set as ranges based on any combination. In addition, the academic documents and patent documents described in this specification are incorporated into this specification as references. Examples

[0055] The present invention will be described in more detail based on the examples below. In addition, the present invention is not limited to these examples.

[0056] <Examples 1 to 4, Comparative Examples 1 to 6> It was studied whether the combination of DHA and lutein has a neuroprotective effect on human neuroblastoma SH-SY5Y cells. Solutions of DHA (manufactured by Sigma-Aldrich) and lutein (manufactured by EXTRASYNTHESE S.A.) dissolved in DMSO (manufactured by NacalaiTesque) were prepared respectively. The method reported previously (Shipley, M.M. et al. J. Vis. Exp. 108, 53193 (2016).; Pezzini, F. et al. Cell. Mol. Neurobiol. 37, 665 - 682 (2017).; Gimenez - Cassina, A. et al. J. Neurosci. Res. 84, 755 - 767 (2006).) was slightly modified to induce the differentiation of human neuroblastoma SH - SY5Y (purchased from ECACC). The cells after differentiation induction had the general morphology of nerve cells. The culture medium used for cultivation is shown in Table 2.

[0057] Wake up SH - SY5Y using Medium 1 and culture it in a CO2 incubator (37 °C, 5% CO2, ambient O2, humidified). After seeding in a 48 - well plate (manufactured by AGC Techno Glass Co., Ltd.) with Medium 1, change to Medium 2 the next day. Change the medium to fresh Medium 2 at a frequency of once every 2 to 3 days. On the 7th day, change to Medium 3 containing DMSO or a DMSO solution of DHA added at the concentrations recorded in Table 1 and / or a DMSO solution of lutein (DMSO concentration is 0.1 wt%), and culture for a further 7 days. Change the medium to the above - mentioned Medium 3 at a frequency of once every 2 to 3 days. On the 14th day, change to Medium 4 containing Rotenone (manufactured by Sigma - Aldrich) added to a final concentration of 0.2 μM, and culture for 48 hours.

[0058] After 48 hours, the culture supernatant was taken, centrifuged at 400×g for 10 minutes, and the supernatant was recovered. PBS (manufactured by Nacalai Tesque) supplemented with 0.1% Triton X-100 (manufactured by Nacalai Tesque) was added to the cells after removing the culture supernatant, and the supernatant was recovered after centrifugation at 2,000×g for 15 minutes. The amount of Lactate Dehydrogenase (LDH) contained in the recovered culture supernatant and the cell lysate was measured using a Cytotoxicity LDH Assay Kit (manufactured by Fujifilm). LDH is an enzyme present in the cytoplasm and normally stays in the cytoplasm. When the cell membrane is damaged, it is released into the culture supernatant. Therefore, the greater the amount of LDH in the culture supernatant, the greater the degree of cell damage. The absorbance at 490 nm was measured using Synergy HT (manufactured by Agilent Technologies), and the average value (N = 3 or 6) of the LDH release level (the ratio of the amount of LDH in the culture supernatant to the total amount of LDH in the culture supernatant and the cell lysate) was calculated. The average value of the LDH release level in Comparative Example 1 (using Medium 3 supplemented with DMSO) was taken as 100%. The results are shown in Table 1. The LDH release level is expressed as a relative value (%) with respect to the LDH release level (100%) in Comparative Example 1.

[0059] [Table 1]

[0060] [Table 2]

[0061] Table 1 shows the values of the LDH release levels in each Example and Comparative Example and the rate of change from Comparative Example 1 (the difference in the LDH release level compared to Comparative Example 1). In addition, for the combinations in the Comparative Examples corresponding to the combinations of DHA and lutein used in Examples 1 to 4, the sum of the rates of change from Comparative Example 1 is also shown. Figure 1 shows the rates of change in Examples 1 to 4 and Comparative Example 6. In addition, for the combinations in the Comparative Examples corresponding to the combinations of DHA and lutein used in Examples 1 to 4, the sum of the rates of change from Comparative Example 1 is also shown. Figure 1In the table, “Comparative Example 2 + Comparative Example 5” is the sum of the rate of change in Comparative Example 2 from Comparative Example 1 and the rate of change in Comparative Example 5 from Comparative Example 1, “Comparative Example 3 + Comparative Example 5” is the sum of the rate of change in Comparative Example 3 from Comparative Example 1 and the rate of change in Comparative Example 5 from Comparative Example 1, “Comparative Example 2 + Comparative Example 4” is the sum of the rate of change in Comparative Example 2 from Comparative Example 1 and the rate of change in Comparative Example 4 from Comparative Example 1, and “Comparative Example 3 + Comparative Example 4” is the sum of the rate of change in Comparative Example 3 from Comparative Example 1 and the rate of change in Comparative Example 4 from Comparative Example 1. “Comparative Example 2 + Comparative Example 5,” “Comparative Example 3 + Comparative Example 5,” “Comparative Example 2 + Comparative Example 4,” and “Comparative Example 3 + Comparative Example 4” refer to the rate of change in the LDH release level in the case of a combination of lutein and DHA, as predicted from the LDH release levels in the case of lutein alone (Comparative Example 2 or 3) and DHA alone (Comparative Example 4 or 5).

[0062] (result) As shown in Table 1, in Comparative Examples 2 to 5 where DHA or lutein was used alone, there was no significant change in the LDH release level compared to Comparative Example 1, and there was basically no protective effect on brain nerve cells. Figure 1 As shown, even if DHA and lutein are combined, in Comparative Example 6 where their weight ratio (DHA / lutein) is outside the range of 10 to 120, there is indeed no significant change in the LDH release level compared to Comparative Example 1, and there is basically no protective effect on brain nerve cells. In contrast, in Examples 1 to 4 where DHA and lutein are used in combination in the above-mentioned weight ratio within the range of 10 to 120, the LDH release level is reduced by more than 5% compared to Comparative Example 1. In addition, as for the rate of change of the LDH release level in Examples 1 to 4, it is significantly lower than the value predicted by the LDH release level in the case of DHA alone or lutein alone (Comparative Example 2 + Comparative Example 5, Comparative Example 3 + Comparative Example 5, Comparative Example 2 + Comparative Example 4 and Comparative Example 3 + Comparative Example 4), so the LDH release level is confirmed to be synergistically reduced. That is, by combining DHA and lutein in a manner within the range of 10 to 120, it is shown that a synergistic protective effect of brain nerve cells is obtained.

[0063] These results indicate that a synergistic effect in protecting brain nerve cells is exhibited by combining lutein and / or its fatty acid esters with docosahexaenoic acid compounds at a specific ratio.

[0064] <Example 5> The effect of a combination of arachidonic acid, DHA, EPA, lutein, and zeaxanthin on memory function in elderly individuals with decreased cognitive function was studied. The memory function was examined using the aforementioned Cognitrax (supplied by Health Solutions Co., Ltd.).

[0065] (Subject) Healthy men and women aged 55 to 79 years with the main complaint of forgetfulness were recruited as subjects. Before the start of the trial, the subjects were administered the MoCA-J (Japanese version of Montreal Cognitive Assessment) as a cognitive function test.

[0066] (Evaluation sample) A test food (soft capsule) containing long-chain highly unsaturated fatty acids (120 mg of arachidonic acid, 300 mg of DHA, and 100 mg of EPA), 10 mg of lutein, and 2 mg of zeaxanthin per 6 capsules was manufactured (weight ratio (DHA / lutein) was 30). In addition, a food with the same appearance but without the above ingredients was manufactured as a control food.

[0067] (Evaluation method) Healthy elderly people with the main complaint of forgetfulness were made to ingest 6 capsules of the test food or the control food per day for 12 weeks. Before ingestion and 12 weeks after ingestion, a comprehensive memory test (verbal memory test and visual memory test) based on Cognitrax was conducted to evaluate the memory function. Among the subjects who completed the trial, 58 subjects (34 in the test food group and 24 in the control food group) who were confirmed to have a decline in cognitive function (MoCA-J score less than 23 points) at the pre-ingestion time point were selected as the analysis subjects. The change amount of the Cognitrax comprehensive memory before and 12 weeks after ingestion and the change amount corrected by the baseline value are shown in Table 3.

[0068] [Table 3]

[0069] (Results) As shown in Table 3, among the subjects with confirmed decline in cognitive function, the group that ingested the test food had a significantly greater improvement in memory function compared to the group that ingested the control food. From these results, it was clarified that combining lutein, zeaxanthin, arachidonic acid, DHA, and EPA in a specific ratio is effective for human memory function.

Claims

1. A composition for protecting nerve cells in the brain, characterized in that, Containing the following components (A) and (B): (A) Lutein and / or its fatty acid esters; (B) Docosahexaenoic acid compounds, and the weight ratio of component (B) (in terms of docosahexaenoic acid) to component (A) (in terms of lutein), i.e., (B) / (A), is 10 to 120.

2. The composition according to claim 1, characterized in that, The docosahexaenoic acid compounds are at least one selected from free docosahexaenoic acid, its salts, and compounds containing docosahexaenoic acid as a constituent fatty acid.

3. The composition according to claim 2, wherein The compounds containing docosahexaenoic acid as a constituent fatty acid are selected from triglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid, diglycerides in which part or all of the constituent fatty acids are docosahexaenoic acid, monoglycerides in which the constituent fatty acid is docosahexaenoic acid, phospholipids in which part or all of the constituent fatty acids are docosahexaenoic acid, glycolipids in which part or all of the constituent fatty acids are docosahexaenoic acid, and alcohol esters of docosahexaenoic acid.

4. The composition according to any one of claims 1 to 3, characterized in that, Component (B) is derived from marine products.

5. The composition according to any one of claims 1 to 3, characterized in that It is an oral composition.

6. The composition according to any one of claims 1 to 3, characterized in that, It is a food or drink.

7. The composition according to any one of claims 1 to 3, characterized in that, It is labeled with at least one function selected from "maintaining cognitive function", "maintaining memory function", "maintaining attention function", "inhibiting brain nerve cell disorders", "protecting brain nerve cells", "guarding brain nerve cells", "reducing brain oxidative stress", "guarding the brain against oxidative stress", and "antioxidation of the brain".

8. The composition according to any one of claims 1 to 3, characterized in that, It further contains zeaxanthin and / or its fatty acid esters, arachidonic acid compounds, and eicosapentaenoic acid compounds.

9. The composition according to claim 8, characterized in that, It is used to improve memory function or inhibit its decline.

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